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Due to the high lipid content, adipose tissue has been challenging to visualize using traditional histological methods. Adipo-Clear is a tissue clearing technique that allows robust labeling and high-resolution volumetric fluorescent imaging of adipose tissue. Here, we describe the methods for sample preparation, pretreatment, staining, clearing, and mounting for imaging.
Adipose tissue plays a central role in energy homeostasis and thermoregulation. It is composed of different types of adipocytes, as well as adipocyte precursors, immune cells, fibroblasts, blood vessels, and nerve projections. Although the molecular control of cell type specification and how these cells interact have been increasingly delineated, a more comprehensive understanding of these adipose-resident cells can be achieved by visualizing their distribution and architecture throughout the whole tissue. Existing immunohistochemistry and immunofluorescence approaches to analyze adipose histology rely on thin paraffin-embedded sections. However, thin sections capture only a small portion of tissue; as a result, the conclusions can be biased by what portion of tissue is analyzed. We have therefore developed an adipose tissue clearing technique, Adipo-Clear, to permit comprehensive three-dimensional visualization of molecular and cellular patterns in whole adipose tissues. Adipo-Clear was adapted from iDISCO/iDISCO+, with specific modifications made to completely remove the lipid stored in the tissue while preserving native tissue morphology. In combination with light-sheet fluorescence microscopy, we demonstrate here the use of the Adipo-Clear method to obtain high-resolution volumetric images of an entire adipose tissue.
Until recently, adipose tissue was conceived of as an amorphous collection of fat cells. Over the past few decades, our understanding has grown more sophisticated, with fat now recognized to be a complex organ containing different types of adipocytes, as well as adipocyte precursors, immune cells, fibroblasts, the vasculature, and nerve projections. Interactions among these adipose-resident cells have pronounced effects on adipose tissue and organismal physiology and pathophysiology1. Although emerging studies have unraveled important molecular mechanisms underlying certain interactions, a more comprehensive understanding requires reliable structural profiling of the entire tissue in three dimensions (3D).
Our current knowledge of adipose tissue morphology is largely based on histological analysis of thin sections (5 μm) with relatively high-magnification imaging (more than 10X)2,3. However, this approach has several significant limitations. First, intricate filamentous structures such as sympathetic nerves and the vasculature, which are known to play important roles in adipose function4,5,6,7, are difficult to evaluate through thin sections. Second, due to its seemingly amorphous shape and the lack of representative structural units to focus on, it is difficult to appreciate adipose tissue structures based only on section staining. Third, adipose tissue has a very high lipid content, creating challenges in obtaining consistent serial sections that are suitable for 3D anatomical reconstruction, a conventional method used to study whole brain morphology8. Given these factors, there is a great need for a whole-mount approach that can provide 3D visualization of an entire adipose depot while still achieving cellular resolution.
3D volumetric imaging of an entire organ is challenging due to the obscuring effects of light scatter. A major source of light scatter in biological tissues comes from lipid-aqueous interfaces. Although the efforts to eliminate scatter by removing lipids have been ongoing for over a century, there have been a large number of recent innovations9. One such newly developed tissue-clearing method is immunolabeling-enabled 3D imaging of solvent-cleared organs (iDISCO/iDISCO+)10,11. However, adipose tissue presents a particular challenge given its high level of lipids, and therefore, additional modifications to the iDISCO/iDISCO+ protocol are required to fully extract the lipids while protecting the tissue from collapsing. The modified protocol we have developed, now called Adipo-Clear, employs methanol/dichloromethane-based delipidation of adipose tissue to achieve optimal transparency suitable for high-resolution volumetric imaging12. Because the delipidation step largely quenches endogenously expressed fluorescent proteins such as GFP and RFP, visualization of such proteins must be achieved by immunolabeling. Overall, this simple and robust protocol can be applied to study tissue-level organization of adipose-resident cells, lineage tracing of adipocyte progenitor cells, and adipose morphogenesis during development.
Animal care and experimentation were performed according to procedures approved by the Institutional Animal Care and Use Committee at the Rockefeller University.
1. Tissue Preparation
Perform standard intracardiac perfusion with ~20 mL of 1x phosphate buffered saline (PBS) at 4 °C until the blood is completely removed from the tissue.
Switch the perfusate to ~20 mL of fixative solution (4% paraformaldehyde (PFA) in 1x PBS) at 4 °C until the neck and tail have significantly stiffened.
CAUTION: PFA is toxic. Avoid contact with skin, eyes, and mucous membrane. Solutions should be made inside a fume hood.
Dissect the fat pads of interest carefully to remove the entire fat pad without damaging it. Use blunt-ended forceps to avoid pinching or squeezing the tissue. Avoid contaminating the dissected tissue with any fur.
Post-fix the tissue in 4% PFA in 1x PBS overnight at 4 °C. For each fat pad, post-fix with ~10 mL of fixation solution in a 15 mL conical tube.
Wash the tissue 3 times (1 h each) with 1x PBS at room temperature (RT).
Store the tissue for short-term (up to 2 weeks) in 1x PBS at 4 °C and protected from light. For long-term storage (e.g., 1-2 years), switch the buffer to 1x PBS with 0.05% sodium azide (as a preservative).
CAUTION: Sodium azide is highly toxic when ingested orally or absorbed through the skin. Concentrated sodium azide solutions (at 5% or greater) should be handled under a fume hood.
2. Delipidation and Permeabilization
Timing: 1-2 days
Prepare 20%, 40%, 60%, and 80% methanol gradient with B1n buffer (Table 1) (v/v), e.g., for 20% methanol/B1n buffer, mix 20 mL of methanol and 80 mL of B1n buffer. Store all buffers at 4 °C. Glycine crystals will precipitate from 60% and 80% methanol/B1n buffers due to saturation. Avoid removing the crystals; use the liquid solution for the following washes.
CAUTION: Methanol is volatile, irritant, and flammable. Avoid skin or eye contact.
Gently remove the hair from the sample under a dissection microscope. Any hair, lint, or debris attached to the sample will cause shadows during imaging. Transfer the cleaned sample into a new tube.
Perform all following steps on ice or at 4 °C unless otherwise indicated. For small samples (e.g., posterior subcutaneous/perigonadal fat pads from young lean mice), use 2 mL microcentrifuge tubes with 1.6 mL of the solution. For large samples or samples with high lipid content (e.g., fat pads from obese mice), use 5 mL tubes with 4 mL of the solution.
Place the tubes containing the samples horizontally on an orbital shaker set at ~ 100 rpm. Make sure the samples can move freely inside the tube.
Dehydrate the sample in a graded series of 20%, 40%, 60%, 80%, and 100% methanol/B1n buffer for the indicated time (Table 2). Minimize the carry-over solution.
Delipidate the sample with 100% dichloromethane (DCM) (3 times), each with the incubation time indicated in Table 2. The sample should sink to the bottom of the tube at the end of the second and third DCM washes. If not, extend the incubation time to ensure full delipidation (e.g., extend from 30 min to 1-2 h).
CAUTION: DCM should be handled inside a fume hood. Use glass containers for storage and microcentrifuge tubes that are made from polypropylene for incubations. The microcentrifuge tubes should not be used for long-term storage of DCM. Petri dishes and serological pipettes that are made from polystyrene are not compatible with DCM. DCM is highly volatile. Transfer the sample into fresh solution quickly to prevent desiccation.
Wash twice with 100% methanol for the indicated time (Table 2).
Optional: If the sample is not well-perfused, the color of hemoglobin from the remaining red blood cells might cause strong autofluorescence. Bleach with 5% H2O2 in methanol (1 volume of 30% H2O2 to 5 volumes of methanol) overnight at 4 °C.
Rehydrate the sample in a reversed methanol/B1n buffer series: 80%, 60%, 40%, 20% methanol/B1n, and 100% B1n buffer (2 times) for the indicated time (Table 2).
Wash the sample with PTxwH buffer (Table 1) for 2 h at RT.
Store the delipidated sample in PTxwH buffer at 4° C (up to 1 year) or proceed immediately to the immunostaining step.
3. Whole Mount Immunostaining
Timing: 8-10 days
Note: All of the following steps should be carried out at RT unless noted otherwise, with shaking and protection from light. For small samples, use 2 mL microcentrifuge tubes with 1.6 mL of solution. For large samples, use 5 mL tubes with 4 mL of solution. It is recommended to first validate the antibodies on small pieces of the tissue or methanol-treated tissue sections.
Dilute the primary antibody in PTxwH buffer to the recommended concentration. Centrifuge the diluted antibody solution at ~20,000 x g for 10 min to prevent introducing antibody precipitates or aggregations.
Incubate the delipidated sample with the primary antibody solution for the indicated time (Table 2).
Wash the sample with PTxwH buffer in a series of incubation steps: 5 min, 10 min, 15 min, 20 min, 1 h, 2 h, 4 h, and overnight.
Dilute the secondary antibody in PTxwH buffer to the recommended concentration. Centrifuge the diluted antibody solution at ~20,000 x g for 10 min to prevent introducing antibody precipitates or aggregations.
Note: To avoid high background caused by tissue autofluorescence, secondary antibodies conjugated with fluorophores that emit light in the red and far-red regions are recommended. Depending on the number of laser lines available on a microscope, up to 3-4 markers can be imaged simultaneously in one sample.
Incubate the sample with the secondary antibody solution for the indicated time (Table 2).
Wash sample with PTxwH buffer in a series of incubation steps: 5 min, 10 min, 15 min, 20 min, 1 h, 2 h, 4 h, and overnight.
Optional: For tissue types that are fragile, fix the stained tissue with 4% PFA in 1x PBS overnight at 4 °C to help preserve tissue morphology.
Note: This step may increase imaging background. It is not necessary to perform this step for adipose tissue.
Wash the sample with 1x PBS in a series of incubation steps: 5 min, 10 min, and 30 min.
Gently remove lint from the sample under a dissection microscope.
4. Tissue Clearing
Optional: Embed the sample in agarose to facilitate sample mounting for light-sheet microscopy.
Note: This step is strongly recommended for adipose tissue to stabilize its shape during imaging.
Prepare the embedding solution with 1% agarose in 1x PBS (w/v). Cool the embedding solution to ~ 40 °C to avoid exposing the sample to excessive heat.
Place the cleaned sample into a mold (e.g., Petri dish or weighing boat) and arrange it into the desired position. Avoid any liquid carryover. Gently pour the embedding solution over the sample. Avoid any air bubbles.
Let the agarose fully solidify at RT. Cut out a block containing the sample.
Note: All of the following steps including overnight incubations should be carried out at RT, with shaking and protection from light. For small samples, use 2 mL microcentrifuge tubes with 1.6 mL of solution. For large samples, use 5 mL tubes with 4 mL of solution.
Dehydrate the sample in methanol gradient with H2O: 25%, 50%, 75%, and 100% (3 times) for the indicated time (Table 2).
Note: Sample can be left overnight at the 100% methanol step.
Incubate the sample in 100% DCM for 1 h with shaking (3 times). Sample should sink to the bottom of the tube at the end of each DCM wash. If not, extend the incubation time to ensure full removal of methanol. Sample can be left overnight at the 100% DCM step.
Incubate the sample in dibenzyl ether (DBE) overnight with mild shaking to achieve refractive index matching.
Note: Sample will eventually become transparent in visible light.
CAUTION: DBE is hazardous. Avoid any contact with skin. Handle it inside a fume hood with double-layered gloves. Discard the gloves as soon as it is contaminated with DBE. Use glass containers for storage and microcentrifuge tubes (polypropylene) for incubations. The microcentrifuge tubes should not be used for long-term storage of DBE. Petri dishes and serological pipettes that are made from polystyrene are not compatible with DBE. Clean any spill with 100% ethanol.
Incubate the sample with fresh DBE with mild shaking for 2 h. Store the sample at RT in the dark or proceed directly to imaging.
Note: Samples are recommended to be imaged within a month. Although certain fluorophores are more stable than others, long-term storage of the samples at this stage is not recommended.
5. Microscopy
Imaging with a light-sheet microscope that is compatible with DBE.
Note: Only objectives that are approved for organic solvent-based imaging and matched with the refractive index of DBE can be used as dipping objectives in DBE-immersed imaging.
Mount the agarose-embedded sample onto a holder that can prevent movement during imaging. Immerse the sample into a DBE-filled chamber.
Collect a z-stack covering the whole sample (e.g., 1-2X magnification) to obtain whole-tissue distribution/structure of the marker of interest.
Switch to an objective with higher magnification (e.g., 4-12X magnification) to zoom in on the regions of interest to image detailed structures.
Note: Collagen is a major contributor to the extracellular matrix of adipose tissue, which surrounds all fat cells13. Collagen has a typical emission spectrum ranging around 400 nm to 550 nm14. Imaging the sample with the 488 laser line (green) and collecting the emitted light at a wavelength range that lies within the emission spectrum of collagen (e.g., 525-550 nm) will provide the tissue autofluorescence signal, which was previously shown to delineate fat cell contour and overall tissue architecture12.
Imaging with an inverted confocal or a two-photon microscope.
Place the agarose-embedded sample into a chamber slide with a glass bottom without touching any plastic parts (or other DBE-compatible imaging chamber that can seal). Make sure DBE does not leak out.
Choose the objectives with longer working distances to achieve deeper penetration.
Note: Imaging should be done with an inverted confocal or two-photon microscope through the glass bottom of the chamber slide. Avoid direct contact between the sample and dipping objectives that are not suggested for DBE-based imaging.
Adipo-Clear prepared whole fat pads can be imaged in 3D to analyze how tissue morphology and cellular interactions are affected in the lean and obese states. This method can be easily applied to analyze general adipose structure by collecting the tissue autofluorescence signal in the green channel. We have previously shown that the autofluorescence signal in adipose overlays favorably with perilipin staining, a commonly used marker to outline mature adipocytes12. For example, scanning a posterior subcutaneous fat pad (psWAT) using a light-sheet microscope with low magnification (1.3X) shows the lobular organization of adipocytes (Figure 1A and B). More detailed information, such as the size of adipocytes, can be revealed by zooming into the regions of interest with higher magnification (4X) (Figure 1C and D).
Adipo-Clear is particularly useful for visualizing filamentous structures such as nerve projections and blood vessels, which are challenging to capture or trace on thin sections. The sympathetic nervous system (SNS) plays a crucial role in controlling lipolysis and thermogenesis in adipose tissue4,5. Imaging a psWAT pad stained with tyrosine hydroxylase (TH), a marker for SNS, reveals the structures that appear as large nerve bundles, blood vessel innervation, as well as dense terminal arborization in the tissue parenchyma (Figure 2A-H). In addition, the TH+ parenchymal projections show regional variation within psWAT, with the inguinal portion having higher density relative to the dorsolumbar portion (Figure 2E-H; Supplementary Movie 1). Our previous work has demonstrated that SNS terminal arborization can be computationally traced and reconstructed using the FilamentTracer tool of Imaris software to assess the density of innervation12.
Adipose tissue is known to be heavily vascularized. The changes in metabolic demands of adipose are often associated with dynamic remodeling of its vasculature6,7. Robust and rapid profiling of whole-tissue vasculature can provide additional unbiased analysis for blood vessel remodeling. Using platelet endothelial cell adhesion molecule (PECAM-1, also known as CD31) as a marker to label blood vessels, we observed that all adipocytes are in contact with the capillaries throughout the whole tissue (Figure 3A-H; Supplementary Movie 2), supporting the high demand for efficient nutrient and oxygen exchange in adipose.
Immune cells are another crucial component of adipose tissue. In the obese state, adipose tissue becomes inflamed, which is accompanied by the infiltration of pro-inflammatory macrophages that form "crown-like" structures surrounding dead adipocytes15,16. Fat pads from obese animals are particularly difficult to clear due to their large size and higher lipid content. However, the extended version of Adipo-Clear (described in Table 2 for large tissue or tissue with high fat content) can achieve consistent clearing of whole high-fat-laden tissue. For example, epididymal fat from a mouse fed with 16 weeks of high-fat diet shows dense "crown-like structures", immunolabeled by CD68, throughout the whole tissue (Figure 4A and B). Importantly, optical sections taken from various positions over the entire depth of the tissue (~4-5 mm) show equally sharp images, demonstrating complete clearing of the tissue (Figure 4C-F).
Figure 1: Analysis of adipose tissue morphology using the autofluorescence signal. All panels are light sheet fluorescence microscopy (LSFM) images of an Adipo-Clear prepared psWAT pad isolated from an 8-week-old C57Bl/6J male mouse housed at RT. The autofluorescence signal is collected by scanning the cleared sample with the green channel. Optical sections (cross-sections from the middle of the sample) of the dorsolumbar region (A) and the inguinal region (B) taken by 1.3X objective. (C, D) High-magnification (4X) optical sections of the boxed regions from A and B. Lymph nodes are indicated by asterisks. Scale bars are indicated in each panel. Please click here to view a larger version of this figure.
Figure 2: 3D imaging of sympathetic innervation in adipose tissue. All panels are LSFM images of a psWAT labeled with tyrosine hydroxylase (TH) (the same sample as in Figure 1). Maximum projections of the reconstructed dorsolumbar region (A) and inguinal region (B) taken by the 1.3X objective. (C, D) Optical sections from the middle of A and B. (E, F) High-magnification (4X) optical sections of the boxed regions from C and D. (G, H) High-magnification optical sections of the overlay between TH (green) and autofluorescence (magenta). Arrowheads indicate distinct patterns of sympathetic innervation: (1) nerve bundle; (2) blood vessel innervation; (3) parenchymal arborization. Lymph nodes are indicated by asterisks. Scale bars are indicated in each panel. Please click here to view a larger version of this figure.
Figure 3: 3D imaging of blood vessels in adipose tissue. All panels are LSFM images of a CD31 labeled psWAT (the same sample as in Figure 1). (A, B) Maximum projections of the reconstructed dorsolumbar region (A) and inguinal region (B) taken by the 1.3X objective. (C, D) Optical sections from the middle of A and B. (E, F) High-magnification (4X) optical sections of the boxed regions from C and D. (G, H) High-magnification optical sections of the overlay between CD31 (red) and autofluorescence (cyan). Lymph nodes are indicated by asterisks. Scale bars are indicated in each panel. Please click here to view a larger version of this figure.
Figure 4: 3D imaging of the "crown-like structures" in adipose tissue. All panels are LSFM images of an Adipo-Clear prepared eWAT pad isolated from a male mouse fed with high fat diet for 16 weeks. The sample was immunolabeled with CD31 and CD68. (A, B) Maximum projections of the reconstructed sample with a total depth of more than 4 mm. (A) X-Y view. (B) Y-Z view. (C-F) Optical sections from the indicated depths in B. Scale bars are indicated in each panel. Please click here to view a larger version of this figure.
Buffer Chemical Final concentration
B1n buffer
Glycine 0.3 M
Triton X-100 0.1% (v/v)
H2O Solvent
Sodium azide (preservative, optional) 0.01% (w/v)
Adjust pH to 7 with NaOH
PTxwH buffer
10x PBS 1x
Tween 20 0.05% (v/v)
Heparin 2 µg/ml
Table 1: List of buffers and solutions. This table contains recipes for the buffers used in Adipo-Clear. For long-term storage of the buffers, it is recommended to add sodium azide as a preservative.
Buffer Small tissue Large tissue (or with high fat content) Temperature
20% methanol/B1n buffer 30 min 1 h 4°C
100% methanol 30 min 1 h 4°C
DCM 30 min 1 h 4°C
DCM 1 h 2-3 h, or Overnight 4°C
Optional: 5% H2O2/methanol Overnight Overnight 4°C
B1n buffer 30 min 1 h RT
B1n buffer Overnight Overnight RT
PTxwH buffer 2 h 2 h RT
PTxwH buffer Storage Storage 4°C
Primary antibody incubation 3 days 4-5 days RT
Secondary antibody incubation 3 days 4-5 days RT
Table 2: Incubation times for delipidation and immunostaining. This table contains incubation times for the delipidation and immunostaining steps of the protocol. The approximate weight of small tissue is < 300 mg.
Supplementary Movie 1: 3D imaging of sympathetic innervation in adipose tissue. Tyrosine hydroxylase (TH) immunostaining of a psWAT sample (same as in Figure 2). The movie shows the fly-through of optical sections (4X) from the dorsolumbar and inguinal regions of psWAT, with a total depth of ~2 mm. TH is shown in green. Autofluorescence is shown in magenta. The region from the dorsolumbar portion appears to have lower SNS density. Please click here to download this file.
Supplementary Movie 2: 3D imaging of the vasculature in adipose tissue. CD31 immunostaining of a psWAT sample (same as in Figure 3). The movie shows the fly-through of optical sections (4X) from the dorsolumbar and inguinal regions of psWAT, with a total depth of ~2 mm. CD31 is shown in red. Autofluorescence is shown in cyan. All adipocytes appear to be closely surrounded by capillaries. Please click here to download this file.
Adipo-Clear is a straightforward and robust method for clearing adipose tissue, which can be easily performed in a regular lab setup. In comparison to other solvent-based clearing methods such as iDISCO/iDISCO+10,11,12, Adipo-Clear is particularly optimized for clearing adipose tissue and other tissue with high fat content. The delipidation step completely removes lipids from adipose, and therefore facilitates immunolabeling throughout the whole tissue and largely minimizes light scatter, allowing end-to-end imaging without any loss of XY resolution. In addition to light-sheet fluorescence microscopes, which provide rapid scanning of large tissue, confocal and two-photon microscopes can also be used to obtain greater resolution and more details.
The methanol/DCM-based delipidation is a critical step. Insufficient delipidation can result in blurred images, especially towards the core of the tissue. It is important to ensure full removal of lipids by observing adipose samples sinking in DCM. The samples that contain a mixture of tissue types (e.g., epididymal fat with epididymis and testis attached) may not fully sink even with extended DCM incubations. However, incubating these samples overnight in DCM should achieve complete delipidation. Due to the denaturation of proteins in these organic solvents, certain antibodies may not be compatible with the delipidation step. Therefore, choosing a suitable antibody becomes another critical step for this protocol. It is recommended to first validate the antibodies on methanol-treated tissue sections or small pieces of tissue processed by Adipo-Clear. Similarly, the compatibility of chemical dyes with methanol/DCM/DBE should also be tested before application.
One limitation of the protocol is the quenching of the endogenously expressed fluorescent proteins. The organic solvent-based delipidation and clearing steps largely denature such proteins. To visualize endogenous fluorescent proteins, antibody labeling needs be employed. The availability of suitable antibody combinations limits the ability to perform highly multiplexed imaging. The current available light-sheet microscopes only allow us to image up to 4 channels.
Adipo-Clear is particularly useful for visualizing filamentous structures and cell populations that have relatively low density in adipose tissue. However, imaging dense signals becomes limited with this approach. When antibodies are used to label dense signals, they can be sequestered by the epitopes located on the surface of the sample, blocking the access to the tissue interior. Therefore, small chemical probes are recommended to stain dense structures. Due to the same issue, the application of Adipo-Clear to brown adipose tissue is limited. Brown fat is an adipose depot with dense structures. In addition to dense blood vessels and nerve innervation, it is also tightly packed with small adipocytes that contain a large number of mitochondria17. When applying CD31 and TH staining with the same procedure as described above to brown fat, only the surface of the sample was labeled (data not shown). Moreover, brown fat produces strong tissue autofluoresence, leading to low signal-to-noise ratio during imaging. It is recommended to cut brown fat into smaller pieces and use chemical probes when possible.
Overall, Adipo-Clear allows simultaneous profiling of multiple structures of interest with high-resolution in whole adipose tissue. Using this approach, one can analyze how structures such as nerve projections, the vasculature, immune cells, and adipocytes interact throughout the whole fat pad. It provides unbiased imaging data by avoiding sectioning or choosing regions of interest. Adipo-Clear can also be applied to study adipose development such as events during early morphogenesis as well as the distribution of adipocyte progenitor and precursor cells in lineage tracing studies. In addition to adipose, Adipo-Clear may also facilitate 3D whole-mount analysis of other tissues that have high lipid content or are surrounded by fat, such as mammary gland and fatty lymph node. This method also offers an opportunity to study the histology of human adipose tissue in physiological and pathological conditions.
The authors have nothing to disclose.
We thank Christina Pyrgaki, Tao Tong, and Alison North from the Bioimaging Resource Center at the Rockefeller University for assistance and support. We also thank Xiphias Ge Zhu for movie editing. This work was supported by the Human Frontier Science Program Organization (PC).
1x phosphate buffered saline Corning 21-040-CV
Paraformaldehyde Sigma Aldrich P6148-1KG
Methanol Fisher Scientific A412SK-4
Triton X-100 Sigma Aldrich X100-500ML
Tween 20 Sigma Aldrich P2287-500ML
Heparin Sigma Aldrich H3393-100KU
Dichloromethane Sigma Aldrich 270991
Hydrogen peroxide 30% Fisher Scientific 325-100
Benzyl ether Sigma Aldrich 108014
Agarose Invitrogen 16500500
Sodium azide Sigma Aldrich 71289-5G
Glycine Fisher Scientific BP381-1
Rabbit polyclonal anti-Tyrosine Hydroxylase Millipore AB152 1:200 dilution
Goat polyclonal anti-CD31/PECAM-1 R&D Systems AF3628 Final concentration of 2 µg/mL
Rat monoclonal anti-CD68, Clone FA-11 Bio-Rad MCA1957 Final concentration of 2 µg/mL
Donkey anti-rabbit IgG (H+L) Alexa Fluor 647 Jackson ImmunoResearch 711-605-152 Final concentration of 5-10 µg/mL
Donkey anti-goat IgG (H+L) Alexa Fluor 568 Invitrogen A11077 Final concentration of 5-10 µg/mL
Donkey anti-rat IgG (H+L) Alexa Fluor 647 Jackson ImmunoResearch 712-605-153 Final concentration of 5-10 µg/mL
Imaging chamber ibidi 80287
Light sheet microscope LaVision BioTec Ultramicroscope II
Imaging software LaVision BioTec Imspector software
Microscopy visualization software Bitplane Imaris
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Supplementary Movie 1


Abbas is an acclaimed photographer who roams around the world covering major political and social events. He has covered wars and revolutions in Biafra, Bangladesh, Northern Ireland, Vietnam, the Middle East, Chile, Cuba, and South Africa during the apartheid. He's been a member of the elite photo agency Magnum since 1981.
The objective of the workshop was to push our creative boundaries as we create a series of photographs that tell a story. I've been shooting for 3 years now and my focus has mostly been on capturing interesting photos that stands on its own and may or may not tell a story. I haven't really tried creating a photo essay – so I guess it's high time.
One by one, the participants lay down 10 to 15 prints of their work on a long table. Abbas would ask everyone to critique, edit, and then sequence each set until it told a story. He'd make the final edit giving us a good peek over his line of thinking. Seeing Abbas edit and sequence my Bad Weather shots was a pretty surreal experience.
The shoot continues… we were taken to a Hindu temple in Little India. It was my first time to shoot in a place like this and got very excited with the barrage of imagery, textures, and ceremonies in the temple. It felt like there was one photographic opportunity after another and you had to keep your eyes peeled to not lose it.
After lunch, the next destination was a Buddhist temple in Toa Payoh. This place was the complete opposite of the Hindu temple, but equally beautiful. The place was spacious and peaceful. The ceremony was quite serene with all the chanting.
Again, it was pretty surreal shooting as Abbas looks on, constantly giving tips on how to see and what to shoot.
I was late for an hour because I joined the Safra Bay Half Marathon Run which started at 5:15am. But still, I couldn't miss this last day… coz as Abbas said, this is the day we start the actual workshop.
We were asked to bring 120 photos from our day-and-a-half shoot. We were grouped in pairs where our partner helped us edit the photos down to 20. These were then projected on the wall where the whole group debated and narrowed it down further. Then all final photos of the participants were printed, laid down on a long table, edited, and sequenced until it gave a pretty good essay on religion / culture in Singapore.
After everything, Abbas gave each of us a signed print of his work and brought out a bottle of Chivas Regal.
Getting to converse with Abbas was a very humbling experience. He kept a low profile and was very fatherly in his approach, but the thoughts he shared in the workshop was absolutely inspiring for me.
Overall, the workshop reintroduced me to the profound side of photography. I've been so focused lately on capturing beauty with a long lens, I forgot to step back and look at the big picture of serious stories happening around. While I still want to pursue portraiture, I have a new-found respect and passion for documentary photography.
In addition to learning from the master himself, I got to meet a group of wonderfully different personalities all sharing the same passion for photography. Big thanks to Nathan, Khanh, Boon, Kim, Fun, Laxmi, and Deanna. I'm looking forward to more photo shoots with you guys!
Melisa – you're awesome. Leonard – you rock, man.
The theme we covered during our 1½ day of shooting was 3 of the prevalent religions in Singapore: Islam, Hinduism, and Buddhism. Abbas has always considered religion as culture rather than faith. With this in mind, we focused not only on the temples and ceremonies, but also on the streets in its periphery – hopefully giving a peek of the people within and outside of their place of worship.
Below are the keepers I got. View these photos larger to see the details, please click here.
View these photos larger to see the details, please click here.
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Sattler College enrolls first-time applicants who present clear and compelling evidence of having completed an academically rigorous program of education at the secondary level, and transfer students with comparable secondary education who have demonstrated, through enrollment in an accredited college or university, that they are capable of succeeding as students in an academically rigorous college. All students enrolled in Sattler College are also required to demonstrate their strong commitment to the fundamental values of Sattler as a Christian college.
Sattler College seeks applications from students who are well-schooled in pre-college or college-level mathematics and life sciences; have a broad and deep knowledge of the history of the United States and the world; have an understanding of the lives of people in societies throughout the world (especially the less developed world) through the study of literature and culture; are able to communicate fluently and effectively in written and spoken English and; demonstrate interest in studying other classical and modern languages. Sattler seeks applications from students who have demonstrated their ability to be both disciplined and self-directed in pursuing their academic goals.
Sattler welcomes applications from students who have acquired their secondary education in a variety of settings, including traditional private and public high schools in the United States or in other countries, as well as students who have received their secondary education at home.
The College encourages applicants to present a wide array of evidence to demonstrate both their academic preparation as well as their commitment to the values of the College. Applicants are encouraged to submit traditional high school transcripts, and results of standardized tests such as the American College Test (ACT); the Scholastic Aptitude Test (SAT), either the general SAT or SAT subject tests; or Advanced Placement Tests. Applicants may also present more qualitative evidence of their academic accomplishments, including statements of the academic program they have followed or portfolios of work accomplished.
All applicants are required to submit a substantial essay that addresses why they are interested in the very particular, focused education that is offered by Sattler College; how their prior academic experience prepares them to succeed at Sattler; and how their decision to pursue a Christian education has influenced their choice of Sattler College.
The College welcomes inquiries from all interested applicants. College faculty and staff are readily available to consult with potential applicants. This openness to inquiries and willingness to consult is particularly important to the application process since Sattler not only accepts but welcomes the presentation of a wide array of evidence in an equally wide array of formats. A student applying to Sattler will not simply fill out forms, but will be asked to reflect on his or her application and present material in forms that best reflect his or her experience.
Students who have submitted applications that are determined by the College to be complete may be asked to participate in a pre-acceptance interview, one that can take place either in person in Boston or through video conferencing.

redpajama_set_name: RedPajamaCommonCrawl


text: Florida man january 1. In an attack on the man who many expect to be his main rival for the 2024 Republican presidential nomination, former president Donald Trump blasted Florida Gov. The zodiac gemstone for Capricorn is ruby. Wesley Scott, 40, was arrested on Friday on an. Thompson. A Florida man was arrested in January after robbing a liquor store using a Spider-Man mask as a disguise, police said. [12] In 2019, a variation of the meme developed on social media, in which people were encouraged to look up "Florida Man" and the date of their birthday, typically finding a bizarre news report involving a 'Florida Man' on that date. Deputies said a man showed up unmasked to a Winn-Dixie Wine and Spirits Store in Casselberry, FL on Jan Bryan Stewart, of Milton Florida, allegedly cut a man with a machete moments after he was heard threatening to kill someone "with kindness," according to the Pensacola and he would have gotten away with it if it weren't for those darned kids and their dog ST. By Stephen Sorace | Fox News. PETERSBURG, Fla. The Pinella Published January 20, 2020 2:13pm EST. A man had taken a cocktail of drugs before being found dead in a hot tub at a glamping site, an inquest heard. He is a man of character and accomplishment. January 21, 2021 at 12:00 pm PST NOW PLAYING ABOVE Man known as 'The Monkey Whisperer' arrested for illegally selling primates (NCD) (NCD) TAMPA, Fla. The. m. They next faced off in the 1969 Gator Bowl. — A St. 2 days ago · Man breaks into Florida rental, takes bath before sleeping in bedroom, deputies say 41 minutes ago No place like dome: $1. He was that state's first solicitor general and sued the Biden. He just told deputies they weren't his. 6, 2021. Naples, Fla. Almost every single day there is a crazy new Florida man headline in the news. . ET LOCATION: Orlando, Florida STADIUM: FBC Mortgage Stadium, home of the UCF Knights TELEVISION: CBS Sports Network The Hula Bowl will take place this year in Orlando, Florida at FBC Mortgage Stadium, home of the UCF Knights, after being previously played in Honolulu, Wailuku, and Halawa, Trump lashes out at DeSantis ahead of potential 2024 clash. Crypto broker Genesis Global Capital is suspending redemptions and new loan originations at its lending business, it said on Wednesday, in the latest sign of the industry fallout from last week's collapse of major crypto exchange FTX. Killed while swimming near Key West, Florida, the man was bitten in the groin area. (WFLA) – A Florida man who made headlines after impersonating a doctor as a teenager and stealing from a patient, landed behind bars again this week. The Sheriff's Office said its Cold Case Unit and the State Attorney's Office is still investigating, as they do with all shootings involving police. Jesse Pack, a 40-year old man from Bunnell, left his front door open and perfectly aligned for officers to see The Florida man who was recorded on New Year's Eve taking a beating from a McDonald's worker after grabbing her over a straw was sentenced to jail last week. WPTV reports Malachi. On January 11, 1994, McMoyler's remains washed up on the beach at Waipio, Hawaii. These conditions include: Laundrie's manner of death was determined to be suicide one month after his remains were found at the Myakkahatchee Creek Environmental Park in Tampa, Florida. 15 On January 10, 1861, . The man was identified as Javon Tyrone Jones. On January 10, 1861, . The former world No. January 9, 2019. 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Arteta is expected to target a new midfielder. Ron DeSantis on Truth Social Thursday night, taking credit for DeSantis . (AP) -- Body camera footage released by authorities showed a man screaming in pain and pleading for help just before a sheriff's deputy shot a Malayan tiger that had grabbed the man's . 1 day ago · Dewhirst earlier this month resigned as solicitor general for Montana, a position he was appointed to in January 2021. These conditions include: In an attack on the man who many expect to be his main rival for the 2024 Republican presidential nomination, former president Donald Trump blasted Florida Gov. Deputies said a man showed up unmasked to a Winn-Dixie Wine and Spirits Store in Casselberry, FL on Jan 2nd. These conditions include: The Collier County Sheriff's Office said the man was seriously injured Wednesday evening when he entered an unauthorized area near the tiger's enclosure at Naples Zoo at Caribbean Gardens. The state seceded from the Union on January 10, 1861, becoming one of the seven original Confederate States, though rejoined the union after the Civil War on June 25, 1868. 1 day ago · The maximum sentence for the obstruction count was 20 years imprisonment. Djokovic, who is Serbian, was deported from Australia in the leadup to the Grand Slam in January after he declined to be vaccinated. The Collier County Sheriff's Office said the man was seriously injured Wednesday evening when he entered an unauthorized area near the tiger's enclosure at Naples Zoo at Florida Man February 1 (2/1) Florida man 'swung a sword' in road-rage attack. The Florida man who hung a giant roll of TP in his front yard just because. The first task for our new Administration will be to liberate our citizens from the crime and terrorism and. In an executive order, the Governor outlined conditions that Kentuckians with at least one of 21 medical conditions, which include cancer, multiple sclerosis, post-traumatic stress disorder, muscular dystrophy or a terminal illness, must meet to access medical cannabis beginning January 1, 2023. 3M hurricane-resistant home hits market in Florida Panhandle DATE: Saturday, January 14, 2023 TIME: 12:00 p. S. -- A Florida man is behind bars after robbing a store disguised as Spiderman. Real estate news with posts on buying homes, celebrity real estate, unique houses, selling homes, and real estate advice from realtor. Bush, he previously served as the 46th governor of Texas from 1995 to 2000. Florida Man is an Internet meme popularized in 2013. Jonathan Bailey, 34-year-old Florida man was arrested Tuesday night outside a Wal-Mart store. 1 was originally barred from the. Florida man attempts to hide underwater from cops, gets arrested after coming up for air, police say. Hubbard was an effective administrator who served 15 consecutive terms while being nationally known as an outspoken segregationalist who sought to keep Dearborn free of the perceived social and political ills of The race is still characterized as a toss-up according to CBS News estimates, with Lawler leading by 3,250 votes, or 1. November 11, 2022. Published: January 21, 2020, 1:16 PM Updated: January 23, 2020, 12:35 PM Tags: Strange Florida , News 6 at Nine Christopher Booth (Suwannee County Sheriff's Office) Garnet is the modern birthstone for the month of January while Emerald is the mystical birth stone (based on Tibetan origin). Fun fact: The birth flower for 1 st January 2001 is Carnation for friendship. A member of the Republican Party, Bush family, and son of the 41st president George H. "I threw pizza at my dad's face because he helped birth me" says Florida man. Chris Lieberman, FISM News. George Walker Bush (born July 6, 1946) is an American politician and businessman who served as the 43rd president of the United States from 2001 to 2009. The suspect laughed and made bird noises as he paced back and forth, completely n**e except for 1 day ago · FILE - Violent rioters, loyal to President Donald Trump, storm the Capitol in Washington, Jan. and he would have gotten away with it if it weren't for those darned kids and their dog A Florida man told police officers he was "teaching it a lesson" when he tried to throw a live alligator he had stolen from a miniature golf course onto the roof of a beachside cocktail lounge . Pete man spent New Year's day in a jail cell after police say he attacked a McDonald's employee for not giving him a straw. These conditions include: A play titled "Florida Man" by Michael Presley Bobbitt premiered July 31, 2019, at New York's Theatre Row Studios. The Florida man who was recorded on New Year's Eve taking a beating from a McDonald's worker January 15, 2021 2:54pm A drunk Florida man was arrested after he was caught operating a floating tiki bar that was reportedly stolen, the US Coast Guard announced . (WTXL) - A Florida man denied syringes were his after jail officials found them in his rectum during a strip search. In 2012, the killing of Trayvon Martin, a young black man, by George Zimmerman in Sanford drew national attention to Florida's stand-your-ground laws, . 1 day ago · FILE - Violent rioters, loyal to President Donald Trump, storm the Capitol in Washington, Jan. The suspect laughed and made bird noises as he paced back and forth, completely n**e except for the mud smeared over his body. A drunk Florida man was arrested after he was caught operating a floating tiki bar that was reportedly stolen, the US Coast Guard announced . After the Florida District 12. 1. Florida Man " refers to an alleged prevalence of male persons performing irrational or absurd actions in the US state of Florida . The Tampa -- A Florida man is behind bars after robbing a store disguised as Spiderman. When the Florida man who was best known for fighting with a giant Easter bunny and arrested after fleeing from an accident, he - Florida Man challenge Find your birthday JANUARY FEBRUARY MARCH APRIL MAY JUNE JULY AUGUST SEPTEMBER OCTOBER NOVEMBER DECEMBER What did the Florida man do on January 16? Florida man Florida Man January 17 (1/17) Wanted Florida man promised officers he would turn himself in when he was finished with his job. Now this has made an interesting searches on Google such as Florida Man January 1, Florida Man March 11, Florida Man March 17 or Florida Man Pastor Florida Man arrested and charged for masturbating on Starbucks patio — for the second time OSCEOLA COUNTY, Fla. Florida Man is an Internet meme first popularized in 2013, [1] referring to an alleged prevalence of male persons performing irrational or absurd actions in the US state of Florida. The court heard how Jai Williams, from Caerphilly in Wales, arrived at Florence. TAMPA, Fla. W. 1 day ago · Speaking at a CNN town hall, former vice-president Mike Pence admitted he was "angered" by a provocative tweet aimed at him by Donald Trump on January 6, 2021, as rioters – some of them . Scott Anthony Massa, 51, allegedly took a bucket of accelerant and attempted to torch his Tampa. Last month, 48-year-old James Mucciaccio of Deerfield Beach, Florida drove his Ferrari 360 off the end of a dock in Palm Beach after a confrontation with a police officer. He is thought to have been killed by an 8-foot (2 m) tiger . He is the right man for the job. The 32-year-old man was arrested early Thursday when Daytona Beach Shores police officers spotted him attempting to throw the gator onto the roof of a cocktail lounge located just off Highway. Florida Man December 1 (12/1) Florida man offered to pay officer with hamburger for o**l s*x. [1] ". These conditions include:. The suspect laughed and made bird noises as he paced back and forth, completely n**e except for 1 day ago · The maximum sentence for the obstruction count was 20 years imprisonment. history. The challenge is simple, just type the words "Florida man" followed by your birthday into Google and see what news stories you get. The United States Census Bureau estimated that the population of Florida was 21,477,737 on July 1, 2019, a 14. National Florida Day is recognized on January 25th each year, and to celebrate, we've compiled a list of some of the most read "Florida Man" stories of the last year. Capitol Police officer was convicted of obstruction of justice on Friday, Oct . He. Florida Man January 1 (1/1) N***d Florida man bites K-9, punches and spits on deputies. Florida and Tennessee would not meet again for fourteen years, the longest gap in the series. A play titled "Florida Man" by Michael Presley Bobbitt premiered July 31, 2019, at New York's Theatre Row Studios. let him out of jail, we need more people like him. Florida Man. Facebook; Jonathan Bailey, 34-year-old Florida man was arrested Tuesday night outside a Wal-Mart store. jamesgilboy. The Florida-based airline cut the most routes from Florida, including flights coming out of Orlando, Fort Lauderdale and Miami. A former U. A Clearwater, Florida man attempted a bold strategy after deputies discovered syringes in his rectum as he was being booked into the Pinellas County Jail. The Seminole County Sheriff's Office said a man was spotted forcing his way into a Winn-Dixie Wine He's a list of 58 of the funniest headlines involving Florida men: "Florida Man Flashes Buttocks at IHOP After Impersonating a Police Officer to Get Free Food" "Florida Man Bites Off Neighbor's Ear Because He Wouldn't Give Him a Cigarette" "Florida Man Attacks Nephew Over Undercooked Noodles" florida man january 1, florida man january 2, florida man january 3, florida man january 4, florida man january 5, florida man january 6, florida man january 7, florida man january 8, florida man january 9, florida man january 10, florida man january 11, florida man january 12, florida man january 13, florida man january 14, florida man january Florida Man nearly set himself on fire while trying to torch his own home Oops: Scott Anthony Massa reportedly took a bucket of accelerant and tried to set his Tampa Bay home on fire Sunday. When Robert Houston, 33 years old and from Holiday in the state Florida Man January 25 (1/25) Florida man known for Easter bunny fight arrested after hit-and-run. com. It has quickly become a huge meme and now there is also a challenge surrounding the concept. Internet users typically submit links to news stories and articles about unusual or strange crimes and other events occurring in Florida , with the headlines . Yasmine James was The U. 2 percentage points, with 98% of jurisdictions reporting results. A fictitious "Florida Man" story. 1 day ago · WASHINGTON (AP) — Attorney General Merrick Garland named a special counsel on Friday to oversee the Justice Department's investigation into the presence of classified documents at former President. Ron DeSantis on Truth Social Thursday night, taking credit for DeSantis' rise to power and blasting him for disloyalty. For instance, my birthday is August 29, so . Pictures and video of this crazy incident can be found around the internet, and you can see that the windshield has definite marks from where he hit it. The January window will provide an opportunity for Arsenal to bring in a couple of new signings as they look to maintain a stunning title challenge. Petersburg Police arrested 57-year old Frank Caponi for offering a hamburger in exchange for o**l s*x to an undercover officer and placing the man under arrest on charges of prostitution, as reported by police spokesperson. St. Which cause it to catch on fire (but that didn't wake him up). PINELLAS COUNTY, Fla. Attorney for the Middle District of Florida said Jimmy Wayne Hammonds, 57, owner of The Monkey Whisperer, illegally sold a capuchin monkey to a buyer in California, A Florida man nearly set himself in flames when he tried to burn his house down, a report said. Agents from the Coast Guard's Key West station. MIAMI (CBSMiami) – A Florida man is in jail charged with aggravated assault with a deadly weapon after he threatened to kill his neighbor "with kindness," because the word "kindness" was. — A Kissimmee pastor who teaches online ministry classes A Florida man nearly set himself in flames when he tried to burn his house down, a report said. Florida was the principal location of the Seminole Wars (1816–1858), the longest and most extensive of the Indian Wars in U. — A Florida man known as "The Monkey. When deputies Florida man January 1, He did something incredible so what did the man do on my birthday? All of a sudden the "Florida Man Headlines" have appeared from nowhere. Four new routes will be added next year, including two non-stop. While the semi-truck drove he bashed the windshield with his fists. Internet users typically submit links to news stories and articles about unusual or strange crimes and other events . The Tampa Bay Times reports that the suspect pulled his car up beside the victim's vehicle and "swung a sword," cutting the other driver's hand A Florida man pelted his girlfriend in the "head and face area" with sweet and sour sauce packets on Sunday because he was angry she had "bought the wrong food from McDonald's," a felony battery . Florida finally beat Tennessee in 1954, winning 14–0 in Knoxville, but Tennessee won the return visit to Gainesville the following year, which was the first time that the teams played at Florida Field. Cops were On August 1, 2020, Florida Man climbed onto the hood of a semi-truck and clung on as it drove 9 miles down the highway. FTX filed for bankruptcy protection on Friday in one of the highest-profile crypto blowups after traders pulled . A more . florida man january 1 ehqybaav bhjmmw dxpz tfgvrbn bbksy kvfeoks ycipmix onxa gmlscu nnhmw


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ShopHome » Products » Carl Dixon – Unbroken
Carl Dixon – Unbroken
Can't Love A Memory
Bowl Me Over
Nothing Lasts Forever
All My Love And Hopes For You
This Isn't The End
Drive Just Drive
Carl Dixon (vocals)
Robert Boebel (guitar)
Thomas Bauer (bass)
Mark Santer (drums)
Dylan Gown (drums)
Produced by: TBD
Engineered by: Robert Bobel, Carl Dixon
Mixed by: Robert Bobel
Mastered by: TBD
Carl Dixon - Unbroken quantity
SKU: AORH00195 Category: CD
Dixon's solo debut 'One' (1993), appears to have been a pretty obvious move for a man of his standing (at least from the European perspective). Carl, born in Sault Ste Marie, Ontario had come to prominence in the early 80s as one of the frontmen for the critically acclaimed Canadian hard rock band Coney Hatch. However, having released three albums between 1982 and 1985, Dixon had become pretty disillusioned with his career at that point; tired of the many frustrations that being on a major label (who don't appear to care) brings on a touring band trying to get noticed. Ironically, as has been shown time and again, North American bands often appear to be curiously shielded from any success they may be achieving abroad. In Coney Hatch's case there was another market out there bursting to embrace them in Europe, if only they'd known about it. After focussing on songwriting for other artists for a while, eventually switching focus from North America, where Grunge was on the brink, to Europe, especially Germany, where Coney Hatch made a much bigger impact, Carl wouldn't fully learn about it only until Coney Hatch' performance at the 2014 edition of Firefest, England. 'One' features fifteen songs, one of them ('One Good Reason') a co-write with Brett Walker, another one ('Taste Of Love') Aerosmith meets Def Leppard flavored. Dixon's first wife Stella ended up adding backing vocals alongside e.g. Mark Santer (Santers) or Mike Shotton (Von Groove). It took a month of recording in four different studios especially focussing on how the guitars sound. The Canadian still is very proud of it and about the fact that people still remember the album. After being with The Guess Who and April Wine for a while and a horrible car crash in Australia in 2008, he's back with Coney Hatch leading to the 2013 record 'Four' and has released a couple of solo records since. Additionally, he's having a brand new solo effort by the name of 'Unbroken' coming out soon on with he collaborated with Frontline's Robby Böbel. After the re-issue of the classic album "One" in October 2019, November sees the release of the first collaboration between Carl and Robert Boebel, a long-time stalwart on the German music scene with bands like Frontline and Phantom V, among many. The result is the new release, Carl Dixon UNBROKEN, a powerfully sung and performed album that reaches the heights of AOR Melodic Rock. Memorable hooks, strong melodies, great guitar playing and production, and drumming performed by Mark Santer (Santers) and newcomer Dylan Gowan (Vesperia, Cardinal Street).
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Hawstead är en by och en civil parish i St Edmundsbury i Suffolk i England. Orten har 334 invånare (2001). Den har en kyrka.

Referenser

Externa länkar
 http://www.britishlistedbuildings.co.uk/england/suffolk/hawstead

Orter i Suffolk
St Edmundsbury District
Civil parishes i Suffolk

redpajama_set_name: RedPajamaWikipedia


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Green Birding Challenge Recap
Written by	Anne Reis
Thank you to everyone who helped make the 5th Annual Green Birding Challenge a success! 19 teams, comprised of both experienced and fledgling birders, participated in this year's challenge and observed over 100 distinct bird species in a fossil fuel free search around the city. Their efforts inspried more than 150 generous donors to pledge $8,000 for the Center's Citzien Science program. Wow! Read on for more fun facts and photos from the day's birding adventure!
Below is a graph of the number of species found by each team and the bonus points earned. This year's Biking Challenge and overall winners with 65 species identified was Pleased to Beat 'Cha! The Spotted Cowbirds and the Common Caws were our Walking and Sitting Challenge winner respectively, while the Satanic Nightjars (it's the name of a real bird!) beat out the competition in the Mini Challenge.
We also made a word cloud based on all of the species observed.
Below are a couple of photos from the event. Be sure to check out our full photo album on Facebook. Please tag yourself in these pictures!
The Satanic Nightjars out on the trail. Photo: Eric Meils
Hoo's Your Daddy and their entire flock! Photo: Anne Reis
Everyone was doing their part for their team. Photo: Eric Meils
We had a great time with you on International Migratory Bird Day and we look forward to seeing you again in 2016!
Photo Credit: Eric Meils
How to Love a River	Native Animal of the Month: Beaver (Castor canadensis) Part 2
Anne Reis
Anne has conducted research on a variety of topics including cranberries, potatoes, wetlands, lichens, tamaracks, and most recently bats. She has a B.S. and M.S. in Horticulture from UW-Madison and a M.S. in Biological Sciences and a GIS Certificate from UW-Milwaukee. Anne is the GIS Specialist at the Center and enjoys reading, mapping, gardening, and spending time outside with her husband and daughter in the Riverwest neighborhood.
Latest from Anne Reis
Research Highlight: Castor canadensis in Riverside Park!
Research Highlight: Migrating Monarchs!
Research Highlight: Much Ado About Mothing
Research Highlight: Share your Urban Ecology Center wildlife sightings - there's an App for that!
Say Hello to the Junco!


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Hock 5
Tigers Surge In Second Half and Roll Into Next Round
OA sophomore Erin Holmberg throws a skip pass over the top of the New Bedford defense during the Tigers' 69-39 win in the Div. 1 South first round. (Josh Perry/HockomockSports.com)
NORTH EASTON, Mass. – The first half was a shootout between the top scorers on each team with New Bedford sophomore Chiron Rose and Oliver Ames senior Kayla Raymond going head-to-head, scoring 15 points apiece. But, it is hard to win a playoff game on the back of one player, even someone like Raymond, the Stonehill-bound Hockomock MVP.
In the second half, the rest of the Tigers took over. Raymond still got her points in the third (before sitting for the entire fourth quarter) but it was OA's depth that turned a six-point lead at the half into a 69-39 victory over the 12th seeded Whalers in the Tigers' first postseason game since moving to Div. 1.
"They came out with fire in their eyeballs," said OA coach Laney Clement-Holbrook. "It was amazing. In the third quarter, we were pushing the pace. We knew that was something that we had to do, we knew we had to run, and I thought the kids did a great job of allowing New Bedford just one shot."
Rose came out firing and got the sizable traveling contingent from New Bedford excited from the opening whistle. She buried a pair of step-back threes and had eight points in the first to help the Whalers get off to a 8-2 start. The Tigers took a few minutes to get going but then Raymond started to take over and helped OA finish the first on a 14-2 run.
She scored on a feed from sophomore Erin Holmberg then added one of her six blocks on the defensive end, which was rebounded by Katie Flynn (10 rebounds) who then fired an outlet pass to Raymond for a transition basket. She assisted on a three-point play for Alex Sheldon (19 points and 13 rebounds), drilled a three and hit a long jumper.
Rose got off to a great start in the second, scoring the first seven points to put New Bedford back in front at 17-16, but Raymond answered right back with layup off a Meg Holleran assist. Abby Reardon (seven points) and Raymond both scored off two more of Holleran's five assists and then Raymond added a layup around the out-stretched arms of New Bedford's 6-foot-2 center Janice Leao (13 rebounds).
A three by Kyara Almeida-Sylvia cut the OA lead to six at the break and a Rose three and a pair from beyond the arc by Monica Eires kept the Whalers in the game, down just 34-29. The Tigers would score the game's next 13 points and in total put together a 25-2 run that finally broke the game open.
"We had a couple match-up situations, but I thought we remedied them in the second half," Clement-Holbrook said. "The unsung heroes like Katie Flynn inside just helping on Leao and just from a collective defensive principle I think that's where we got the points from. We shut them down and just ran the floor."
Freshman Caroline Flynn got it started with a drive to the rim and she added three assists in the second half. Holmberg scored all eight of her points in the third quarter, finding leak-out opportunities for transition buckets. Sheldon was also a force in the paint, scoring 11 of her 19 points in the second half.
Clement-Holbrook said of Sheldon's contributions, "She was absolutely, positively key in all this because everyone's trying to take away Kayla and the fact that we could get the ball inside…we're just sending shuffle cuts and good movement and that allowed Alex to get the ball and she's very physical and she's very strong. She came to play today."
Raymond, who finished with a line of 23 points, seven rebounds, six blocks, four assists and three steals, was still doing her thing in the third, capping the quarter by picking Rose clean at one end and swooping in for a finger roll at the other. The New Bedford guard, who was held to three points in the second half, was left with a wry smile on her face as she looked up to the rafters following the steal.
"Rose in the first half, she was money and we gave her a little too much room to wiggle," Clement-Holbrook explained. "In the second half we did a much better job. When we switched Kayla on her, I thought Kayla's length really bothered her."
OA led 51-31 after three quarters. The Tigers scored one fewer point in the third than they had in the entire first half. That continued in the fourth quarter as Tate Hadges came off the bench and scored all six of her points, Sheldon continued to control the paint and a trip to the Div. 1 South quarterfinal was confirmed.
When it was mentioned to Clement-Holbrook that the move to Div. 1 seems to be going well, she smiled and replied, "Currently, but we'll see how these things go."
Oliver Ames (16-5) will travel to No. 4 seed Bridgewater-Raynham in the quarterfinal round on Saturday, barring a change because of the impending storm.
Josh Perry
Josh Perry is the Managing Editor of HockomockSports.com. He can be contacted at JoshPerry@hockomocksports.com and followed on Twitter at @Josh_Perry10.
Latest posts by Josh Perry (see all)
Girls Basketball Photos: Oliver Ames vs. Franklin - 01/17/2020
Quinn Sets Career-High as Franklin Rolls at OA - 01/17/2020
Boys Hockey Photos: King Philip vs. North Attleboro - 01/16/2020
Abby Reardon Alex Sheldon Caroline Flynn Erin Holmberg Katie Flynn Kayla Raymond Laney Clement-Holbrook Meg Holleran New Bedford Oliver Ames Tate Hadges
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No more interleague games against the American League for a year. We're finished with different teams, different pitchers, different hitters, different rules, different parks, different hotels, different time zones and different running water.
The numbers aren't pretty, 4-11 record, .220 batting average and 57 runs scored. Only the Padres (3-15) had a poorer record.
Speaking of running water, the NL took a total bath: 102-149 (NL East 34-44, NL Central 41-51, NL West 27-54). The AL held the upper hand all around: .275-.251 average, 1,247-1,010 runs, 276-251 home runs and 3.69-4.56 ERA. For the record, there is one game left on the schedule, a June 26 rain out of the Yankees at Pittsburgh that is now set for July 10.
So, so long to the AL and back to the NL. 19 of the next 25 games are within our division. Three months remain in the 2008 marathon season and every game and series is big.
Despite losing five straight series, we will still be in first place tomorrow morning. In case you are wondering, the Phillies have been leaders on July 1 in seven previous seasons: 1950, 1976, 1978, 1993, 1995, 2001 and 2004.
Last time we won a series was in Atlanta where we begin a three-game set tomorrow night. We swept three from the Braves there during the first weekend of this month. I'm sure the Braves haven't forgotten.
Appreciate the postings following Saturday's Phillies Insider. Jenn added a cool suggestion for what to do with the bats, peg-leg for pirates.
"Build a bandwagon that we'll soon be jumping off" was another posted comment. Another clever idea but don't leap.
1932 Chuck Klein hits 23rd and 24th homers in 9-3 win over Brooklyn at Baker Bowl, the most home runs at that time by the end of June by any National League player.
1938 New York Giants romp, 14-1, over the Phillies in final game at decaying Baker Bowl before 1,500 fans. On July 4, the Phillies moved to Shibe Park, home of the Philadelphia A's.
1967 Cookie Rojas pitches scoreless ninth inning in 12-3 loss to the Giants, 2nd game. Rojas has now filled all nine positions since joining the Phillies in 1963, and played eight different positions this season.
In case you haven't noticed, we are scuffling. The offense has gone south since the 20-run win in St. Louis on Friday the 13th.
**Burn them in a bonfire.
**Invite a colony of termites for a feast.
**Sell them to a toothpick factory.
**Dip them in The Bull's spicy barbeque sauce.
**Stick them in a freezer to make them harder.
**Drape them with flowers as a sign of affection.
**Send them back to the forest.
**Grind them into kitty litter.
Charlie Manuel was the pre-game story yesterday as he drastically changed the lineup on his V-8 Machine. While the Phillies still struggled with runners on base, the slump, streak, nose-dive, decline–whatever you want to call it–came to an end.
The game story belonged to Kyle Kendrick and Chase Utley. Kyle dominated the A's for eight innings, throwing 71 strikes out of 113 pitches. He demonstrated what command can do, especially pitching inside to a lineup dominated by left-handed hitters. Going into the game, LH hitters were batting .328 against him.
Utley broke out of his mixup with a four-hit game–single, double, triple, single in that order. When asked how worried he was over Utley's recent struggles, Manuel said, with a grin, "I was scared to death," as Todd Zolecki reported in this morning's Inquirer.
Eric Bruntlett, Mr. Versatile, made his first start at first base last night. He borrowed Rich Dubee's first baseman's mitt. Frank Coppenbarger, he of equipment fame, needs to order Eric's own glove.
So, winning two of three comes down to this afternoon's rubber game. Next, three games in Texas (different time zone) and three games next week in Atlanta (another different time zone).
Rollins ss; Utley, 2b; Burrell, lf; Howard, 1b; Dobbs, 3b; Victorino, cf; Werth, rf; Jenkins, dh; Ruiz, c. Adam Eaton on the mount.
Lehigh Valley OF Chris Snelling has been placed on the DL (left knee inflammation) . . . Clearwater RHP Kris Benson has been promoted to LV . . . Clearwater RH reliever Brett Harker is moving up to Reading . . . Reading LH reliever Jason Kershner was released.
OF Zack Collier was named to the USA TODAY High School All-Star team . . . SS Anthony Hewitt was selected to the second team. Hewitt and Collier were the first two players we selected in the draft earlier this month.
In response to a comment yesterday, Doug Mansolino is wearing #53.
1987 RHP Steve Bedrosian ties major league record (Sparky Lyle, Yankees, 1973) with his 11th saved in last 11 appearances, 5-2, over Mets.
When things are going good, you can feel it. When things aren't going good, you can feel it.
A key base, a big pitch, a defensive play….all part of winning games. Right now, it just isn't happening. Every little thing is magnified.
*Pat Burrell's home run last night gave us a lead for the first time since the Monday game against Boston. That's hard to believe.
*Jamie Moyer was brilliant. Yet, one pitch did him in.
*Trailing, 3-2, in the 8th, J. C. Romero gave up a two-run, opposite-field home run to a left-handed batter who has been scuffling. It was the first homer by a lefty off J. C. since April 2006.
It is a case of hanging in there and battling until things get turned around.
RHP Kris Benson made his second rehab start for Clearwater last night against Vero Beach: 4.2 innings, 7 hits (6 singles), 1 run (top of 5th inning), 1 walk, 3 strikeouts. "Command was better with all his pitches and he was down in the zone more often," reported Steve Schrenk, pitching coach.
Benson took the 2-1 loss. Clearwater had trouble hitting with runners on base. Three times with a man on third and one out, the next hitter struck out. They must be watching the big club.
Came across a great story on the Oakland A's website. Ryan Howard and A's reliever Brad Ziegler were roommates in Clearwater four years ago. Ziegler was originally drafted by the Phillies.
1986 Phillies release Steve Carlton. In 15 years with the Phillies, he won 241 games, started 499 and struck out 3,031, all club records. He won 10 or more games for 13 consecutive seasons, another Phillies record.
Still remember the night very well. Went into Bill Giles' office well after the game was over. Bill still had tears in his eyes because he had to deliver the news to Lefty.
Three teams in the majors have a winning record on the road and the Phillies (20-17) are one of the trio. The others: St. Louis (21-16) and Angels (24-12).
Since 2004, only the Angels (202) and Yankees (194) have more wins on enemy turf or sod than us (192).
When Jamie Moyer starts, we're 10-5.
Hopefully, these numbers bode well to end the gloomy five-game spiral.
Against the A's, we're 3-3, winning two of three at home in 2003 and losing two of three in the McAfee Coliseum in 2005.
The rest of the Phillies hitters have never faced the A's.
Charlie Manuel can use the DH tonight and for the next five days. The A's are starting a RHP, LHP and RHP these three games.
Most likely options: Burrell, Dobbs, Victorino, Werth, Bruntlett, Jenkins. Sorry, catching instructor Mick Billmeyer. We're not activating a .247 minor league hitter who hasn't batted in 15 years!
1994 Phillies sign two free agents, LHP Fernando Valenzuela and INF Randy Ready. Sorry, Fernando and Randy, but history is kind of bleak for this date. Perhaps something historic will occur tonight. Ending a five-game losing streak isn't historic, but at this point, we'll take it!
Two weeks ago we swept the Braves in Atlanta. What a high.
Now, we've dropped five in a row. What a low.
Someone once told me: "Don't get too low when the going is bad and don't get to high when the going is good." That someone? The late Gene Mauch.
Manuel's V-8 Machine has been misfiring. But, it will get going again. This is too good an offensive club to keep sputtering. Hitting is contagious which, I suppose, means non-hitting is the same.
Bottom line: we're in first place as we head for a tough nine-game road trip through Oakland, Arlington and Atlanta.
In looking over various message boards, some fans are for it and others oppose it.
One concern is quality of opponent. In our case, we have a very difficult interleague schedule. But, that's the way it goes. "There's no crying in baseball," Tom Hanks, as manager, said in the movie, League Of Their Own.
Still like seeing other players from teams in the other league, although Vladimir Guerrero can stay in California next time.
During the month of June, the Phillies averaged more than 44,500 in their 11 home dates. The total attendance was 489,879.
Ten of the games were sold out, including the last nine.
Appreciate the most recent comments.
Will get to searching for pitching in a later Phillies Insider. Just keep in mind discussing certain enemy pitchers is tampering and baseball has a huge fine for tampering.
1971 RHP Rick Wise becomes first pitcher in major league history to hit two home runs while pitching a no-hitter, 4-0, in Cincinnati. Wise is currently the pitching coach for the Lancaster (PA) Barnstormers in the Atlantic League. Von Hayes is the manager.
1973 Ken Brett sets a major league record for pitchers by hitting a home run in his fourth straight game, a 7-2 win in Montreal.
1978 Phillies beat Cubs twice by 6-1 scores at Veterans Stadium to take over the NL East lead, a lead they held for the rest of the season.
1986 Phillies bomb Cubs, 19-1, at the Vet, setting modern NL record with 15 extra-base hits and a new club record with 11 doubles. HRs by Juan Samuel (2), Mike Schmidt and Milt Thompson.
Fortunately, or unfortunately–depending upon how readers view this blog–Phillies Insider will be blank as I'm going to be away for the weekend series against the Angels.
Thanks for checking out Phillies Insider. Appreciate your passion.
Rollins, ss; Victorino, cf; Utley, 2b; Howard, 1b; Burrell, lf; Jenkins, rf; Dobbs, 3b; Ruiz, c; Kendrick, p.
24 hours after the Phillies had taken the NL lead in runs scored, LHP Jon Lester and his bullpen mates tossed a shutout, setting up the rubber game this afternoon.
Phillies offense has been inconsistent at times but credit last night goes to Lester, who was involved in his fourth scoreless outing of the season.
60 years ago today Robin Roberts made his major league debut, as was mentioned in this space yesterday.
A young man the National League believes has a date with pitching stardom . . . a spunky, right-hander with a violent fast ball, a crackling curve and though still a raw rookie in a professional sense, has no fear of any hitter he faces. Signed out of Michigan State, this imperturbable youngster debuted in the Inter-State League with Wilmington last season, fanned an average of nine per game, won nine of 10 decisions and was brought up in mid-June. Pitched 10 complete games, won 7, lost 9; fanned 84 and in all worked in 20 games. Beat Cincinnati three out of three; also scored wins over Boston, Chicago and Pittsburgh.
Chase Utley continues leading all NL players, which is great. Matter of fact, his lead over the second-place vote-getter (Chipper Jones) increased.
Pat Burrell moved from ninth to sixth and Jimmy Rollins from fifth to fourth. But, Ryan Howard, dropped a notch to fifth place. Rollins had the second most votes among shortstops over the past week which is good news.
We need to get behind our players. In-park voting ends Saturday but online voting continues until 1:59 p.m. July 2. Fans can vote up to 25 times on phillies.com.
OF Greg Golson (Reading) has been placed on the DL with a left wrist strain . . . RHP Tyson Brummett (Clearwater) has been promoted to Reading . . . OF Michael Taylor and RHP Drew Naylor (Lakewood) have been moved up a level to Clearwater. Each was having a banner year with the BlueClaws.
Rollins, ss; Victorino, cf; Utley, 2b; Howard, 1b; Burrell, lf; Werth, rf; Feliz, 3b; Ruiz, c; Moyer, p.
Citizens Bank Park was really alive last night, electricity as in a playoff game. Jimmy Rollins got the juices going with his lead off home run, his 28th, which is twice as many as the Phillies previous record holder, who happens to be Juan Samuel.
Ryan Howard is getting hot. The two MVPs were the catalysts and Cole Hamels did the job on the mound.
Something different seems to happen in every baseball game. Triples by Howard and Pat Burrell in the same game…..that's something different. Howard now has more triples than J-Roll, 2-1. That's something different.
Thanks for the feedback on Juan Samuel. He is a special person, on top of being a very good ballplayer.
Sammy's evening yesterday started with an appearance on Comcast SportsNet's Daily News Live from the field. It was followed by a press conference in the Media Room, 30 minutes of autographs in the Hall of Fame Club, Phillies-pre-game radio show with Jim Jackson in the Main Concourse, in-game Phillies TV and Spanish Radio.
Wherever he went yesterday, people knew him…..media, players, coaches, managers, game day employees, club officials and the fans. He was smile city last night.
He'll be on the mound again tomorrow, wearing that familiar number 36. The motion will still be fluid but the speed won't register on the radar gun.
"He" is Hall of Famer Robin Roberts. To commemorate the 60th anniversary of his major league debut (June 18, 1948), Robbie will throw out the first ball.
Robbie was a pitcher with a blazing fast ball, awesome curve and pinpoint control. His motion was as fluid and smooth as any who hurler who has graced this game. He never backed hitters off the plate and paid for it by allowing a ton of home runs (505). In the end, he won 286 games and made it to Cooperstown, NY.
To say he was a workhorse would be a gross understatement. For six straight seasons starting in 1950, he never threw under 304 innings. He would have set insurmountable pitch-count records but pitch counts didn't exist back then, thank goodness.
At one point in his career, he had 28 consecutive complete games (8/25/52 to 7/5/53). Robbie led the majors with 33 CG in 1953. The game has changed. The Diamondbacks staff led the NL last season with 7, a month's worth for Robbie.
Robbie, now 81, will be honored as part of Toyota Alumni Night set for Saturday, August 9. Family, friends and ex-teammates will join Phillies Alumni that night. All fans will receive a commemorative print of Roberts, thanks to Toyota.
1930 RF Chuck Klein hits in 26th straight game (.432), May 18-June 17, a new modern club record. He hit in 26 in a row again the same season (.434), July 12-August 13.


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Miscanthus – for wildlife and game cover
There have been environmental studies done in the UK which show that Miscanthus provides excellent habitat for a variety of wildlife including nesting birds. To some extent Miscanthus New Zealand Ltd (MNZ) has discounted these reports because what is good in the UK is not necessarily good in NZ. But innovative people in NZ, almost all of whom come from a UK background, have been purchasing and planting Miscanthus specifically to provide safety and cover to game birds such as pheasants.
It was news to MNZ that there is a thriving commercial game bird shooting industry in New Zealand but at least three such businesses have now planted their own Miscanthus and seem very happy with the result.
Apparently the Miscanthus allows the birds to be protected from predators such as hawks while also providing physical shelter from the elements – as has been discussed to some extent in relation to agricultural fauna in the article 'Miscanthus – for farm shelter'.
That got MNZ staff thinking because it began to explain what has been observed over several years of trial evaluation. Firstly, Miscanthus stands do not provide good habitat for rodents. Apart from the compact bases of the plants, the lack of any seed means that there is nothing in such stands for them to eat. MNZ has not observed any evidence of rodent presence in New Zealand Miscanthus stands. But it is also obvious to anyone who observes wildlife that Miscanthus provides a good refuge, favoured by several species including indigenous fauna. But does it harbor any problem species?
When Miscanthus is first getting established, rabbits can make their presence felt by browsing on newly emerging leaves. But this damage tends to be localised and affects only individual plants or small groups. In NZ, this has only once proved to be a problem when there was a large uncontrolled rabbit population and a very small Miscanthus "trial", surrounded by well-grazed pasture. With no rabbit control at all in this trial, the heavy pressure from the large rabbit population virtually eliminated the Miscanthus plantlets. In more typical situations, MNZ has observed that Miscanthus quite quickly outgrows the rabbit pressure and then creates conditions that are unsuitable for rabbits.
MNZ was originally curious and concerned about the possibility of wild pigs rooting up the rhizomes to eat them. However despite wild pigs being common in many areas around the world where Miscanthus is grown, there have to our knowledge been no reports of any damage to Miscanthus stands at all.
MNZ has also observed that at one or two sites, Pukeko can be somewhat of a problem at the establishment stage in the same way that they can be a problem with the establishment of maize – i.e. by pulling out the newly emerging plantlets. But in most cases the emerging Miscanthus simply outgrows the Pukeko predation. On a positive note, once the Miscanthus stand is established, it has been observed that Pukeko favour it as a refuge and we suspect that they may use it as cover for breeding.
Quail have also been commonly observed in and around Miscanthus stands and although no studies have been done, they may also be using the stands for refuge and perhaps for breeding.
As mentioned above, pheasants definitely benefit from having Miscanthus for cover and owners of game bird shooting operations with whom MNZ has spoken have indicated that their planting of Miscanthus was a sound decision.
Deer are the other wildlife that can be found in Miscanthus stands. MNZ has seen deer (indigenous) in and near a Miscanthus stand in Illinois where the farmer regarded them fondly and was not aware of any negative impact on the Miscanthus from the presence of the deer.
In New Zealand, the MNZ Miscanthus stand near Helensville has a significant population of fallow deer. They use the stand for cover once the Miscanthus has grown tall enough to hide them. If you walk through the stand in late December or January, you will find that in every little stand gap where there has been sufficient light to stimulate any grass growth, deer droppings are plentiful. When the stand is harvested six to nine months later, the deer congregate in the remaining stand until it is harvested to the point where there insufficient area left for them to hide. They then take off to the adjoining pine forest, where they remain until the re-growing Miscanthus in Spring is tall enough to again provide them with cover.
What this all means is that Miscanthus stands stimulate wildlife in general but do not support rodents. As Lincoln University work has shown, this includes indigenous fauna habitat, such as the skinks in Canterbury. If funding were available, formal research in commercial Miscanthus planted throughout New Zealand, could be expected to identify even more positive wildlife benefits from the establishment of such stands.
The next article should be on the value of Miscanthus for stabilising and otherwise improving soils.
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