Name: visualization of 3d white adipose tissue structure using whole-mount staining
Uploaded: 2018-11-17T14:45:00
Description: Watch this Scientific Journal Video about Visualization of 3D White Adipose Tissue Structure Using Whole-mount Staining at JoVE.com

This work was funded by grants from the Natural Sciences and Engineering Research Council (NSERC) of Canada, Pilot and Feasibility Study Grant of Banting &#38; Best Diabetes Centre (BBDC), the SickKids Start-up Fund to H-K.S., Medical Research Center Program (2015R1A5A2009124) through the National Research Foundation of Korea (NRF) funded by the Ministry of Science, ICT, and Future Planning to J-R.K.

All experimental animal protocols were approved by the Animal Care Committee of The Center for Phenogenomics (TCP) conformed to the standards of the Canadian Council on Animal Care. Mice were maintained on 12-h light/dark cycles and provided with free access to water and food. 7 month old C57BL/6J male mice were used in the whole-mount staining experiment.
NOTE: Sections 1 to 2 are in chronological order, with section 3 being an optional step right after section 1. Section 4 can be performed t...

<ol>
	<li>Sung, H. K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Adipose+vascular+endothelial+growth+factor+regulates+metabolic+homeostasis+through+angiogenesis.">Adipose vascular endothelial growth factor regulates metabolic homeostasis through angiogenesis.</a> Cell Metabolism. 17, 61-72 (2013).</li><li>Greenberg, A. S., Obin, M. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Obesity+and+the+role+of+adipose+tissue+in+inflammation+and+metabolism.">Obesity and the role of adipose tissue in inflammation and metabolism.</a> American Journal of Clinical Nutrition. 83, 461-465 (2006).</li><li>Martinez-Santiba&#241;ez, G., Cho, K. W., Lumeng, C. N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Imaging+White+Adipose+Tissue+With+Confocal+Microscopy.">Imaging White Adipose Tissue With Confocal Microscopy.</a> Methods in Enzymology. 537, 17-30 (2014).</li><li>Laforest, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Comparative+analysis+of+three+human+adipocyte+size+measurement+methods+and+their+relevance+for+cardiometabolic+risk.">Comparative analysis of three human adipocyte size measurement methods and their relevance for cardiometabolic risk.</a> Obesity (Silver Spring, MD). 25 (1), 122-131 (2017).</li><li>Berry, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Imaging+of+adipose+tissue.">Imaging of adipose tissue.</a> Methods in Enzymology. 537, 47-73 (2014).</li><li>Kim, K. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Intermittent+fasting+promotes+adipose+thermogenesis+and+metabolic+homeostasis+via+VEGF-mediated+alternative+activation+of+macrophage.">Intermittent fasting promotes adipose thermogenesis and metabolic homeostasis via VEGF-mediated alternative activation of macrophage.</a> Cell Research. 27 (11), 1309-1326 (2017).</li><li>Cho, C. H., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Angiogenic+role+of+LYVE-1-positive+macrophages+in+adipose+tissue.">Angiogenic role of LYVE-1-positive macrophages in adipose tissue.</a> Circulation Research. 100 (4), e47-e57 (2007).</li><li>Lee, J. H., Yeganeh, A., Konoeda, H., Moon, J. H., Sung, H. K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Flow+Cytometry+and+Lineage+Tracing+Study+for+Identification+of+Adipocyte+Precursor+Cell+(APC)+Populations.">Flow Cytometry and Lineage Tracing Study for Identification of Adipocyte Precursor Cell (APC) Populations.</a> Methods in Molecular Biology. 1752, 111-121 (2018).</li><li>Chi, J., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Three-Dimensional+Adipose+Tissue+Imaging+Reveals+Regional+Variation.">Three-Dimensional Adipose Tissue Imaging Reveals Regional Variation.</a> in Beige Fat Biogenesis and PRDM16-Dependent Sympathetic Neurite Density. Cell Metabolism. 27 (1), 226-236 (2018).</li><li>Jiang, H., Ding, X., Cao, Y., Wang, H., Zeng, W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Dense+Intra-adipose+Sympathetic+Arborizations+Are+Essential+for+Cold-Induced+Beiging+of+Mouse+White+Adipose+Tissue.">Dense Intra-adipose Sympathetic Arborizations Are Essential for Cold-Induced Beiging of Mouse White Adipose Tissue.</a> Cell Metabolism. 26 (4), 686-692 (2017).</li><li>Cao, Y., Wang, H., Wang, Q., Han, X., Zeng, W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Three-dimensional+volume+fluorescence-imaging+of+vascular+plasticity+in+adipose+tissues.">Three-dimensional volume fluorescence-imaging of vascular plasticity in adipose tissues.</a> Molecular Metabolism. , (2018).</li><li>Cao, Y., Wang, H., Zeng, W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Whole-tissue+3D+imaging+reveals+intra-adipose+sympathetic+plasticity+regulated+by+NGF-TrkA+signal+in+cold-induced+beiging.">Whole-tissue 3D imaging reveals intra-adipose sympathetic plasticity regulated by NGF-TrkA signal in cold-induced beiging.</a> Protein &amp; Cell. 9 (6), 527-539 (2018).</li><li>Renier, N., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Mapping+of+Brain+Activity+by+Automated+Volume+Analysis+of+Immediate+Early+Genes.">Mapping of Brain Activity by Automated Volume Analysis of Immediate Early Genes.</a> Cell. 165 (7), 1789-1802 (2016).</li><li>Renier, N., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=iDISCO:+a+simple,+rapid+method+to+immunolabel+large+tissue+samples+for+volume+imaging.">iDISCO: a simple, rapid method to immunolabel large tissue samples for volume imaging.</a> Cell. 159 (4), 896-910 (2014).</li><li>Kretzschmar, K., Watt, F. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Lineage+tracing.">Lineage tracing.</a> Cell. 148 (1-2), 33-45 (2012).</li><li>Vorhagen, S., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Lineage+tracing+mediated+by+cre-recombinase+activity.">Lineage tracing mediated by cre-recombinase activity.</a> Journal of Investigative Dermatology. 135 (1), 1-4 (2015).</li><li>Berry, R., Rodeheffer, M. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Characterization+of+the+adipocyte+cellular+lineage+in+vivo.">Characterization of the adipocyte cellular lineage in vivo.</a> Nature Cell Biology. 15 (3), 302-308 (2013).</li><li>Muzumdar, M. D., Tasic, B., Miyamichi, K., Li, L., Luo, L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+global+double-fluorescent+Cre+reporter+mouse.">A global double-fluorescent Cre reporter mouse.</a> Genesis. 45 (9), 593-605 (2007).</li><li>. iDISCO+ protocol Available from: <a target="_blank" href="https://www.idiscodotinfo.files.wordpress.com/2015/04/whole-mount-staining-bench-protocol-methanol-dec-2016.pdf">https://www.idiscodotinfo.files.wordpress.com/2015/04/whole-mount-staining-bench-protocol-methanol-dec-2016.pdf</a> (2016)</li><li>Jensen, E. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Quantitative+analysis+of+histological+staining+and+fluorescence+using+ImageJ.">Quantitative analysis of histological staining and fluorescence using ImageJ.</a> The Anatomical Record (Hoboken). 296 (3), 378-381 (2013).</li><li>Papadopulos, F., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Common+tasks+in+microscopic+and+ultrastructural+image+analysis+using+ImageJ.">Common tasks in microscopic and ultrastructural image analysis using ImageJ.</a> Ultrastructural Pathology. 31 (6), 401-407 (2007).</li><li>Stanly, T. A., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Critical+importance+of+appropriate+fixation+conditions+for+faithful+imaging+of+receptor+microclusters.">Critical importance of appropriate fixation conditions for faithful imaging of receptor microclusters.</a> Biology Open. 5 (9), 1343-1350 (2016).</li><li>Spalteholz, W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> U&#776;ber das Durchsichtigmachen von menschlichen und tierischen Pra&#776;paraten und seine theoretischen Bedingungen, nebst Anhang: U&#776;ber Knochenfa&#776;rbung. , (1914).</li><li>Girkin, J. M., Carvalho, M. T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+light-sheet+microscopy+revolution.">The light-sheet microscopy revolution.</a> Journal of Optics. 20 (5), 053002 (2018).</li><li>Susaki, E. A., Ueda, H. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Whole-body+and+Whole-Organ+Clearing+and+Imaging+Techniques+with+Single-Cell+Resolution:+Toward+Organism-Level+Systems+Biology+in+Mammals.">Whole-body and Whole-Organ Clearing and Imaging Techniques with Single-Cell Resolution: Toward Organism-Level Systems Biology in Mammals.</a> Cell Chemical Biology. 23 (1), 137-157 (2016).</li></ol>

Although conventional techniques such as histology and cryosection offer benefits for observing intracellular structure, whole-mount staining provides a different perspective in adipose tissue research, which enables 3D visualization of cellular architecture of minimally processed tissue.
In order to successfully perform whole-mount staining, the following suggestions should be taken into consideration. Different adipose tissue depots can yield various immunostaining results; thus, the type of...

Adipose tissue is an essential organ for energy storage and is characterized by dynamic remodelling and nearly unlimited expansion1. In addition to energy homeostasis, adipose tissue also plays an essential role in hormone secretion of over 50 adipokines to modulate whole-body metabolic function2. Adipose tissue has a diverse architecture comprising of various cell types including mature adipocytes, fibroblasts, endothelial cells, immune cells, and adipocyte progenitor cells3. Recent studies have shown that obesity and other metabolic dysfunction can significantly alter adipose tissue function and its microenvironment, which includes but is not limited to enlargement of adipocytes, infiltration of inflammatory cells (e.g., macrophages), and vascular dysfunction3.
Conventional morphological techniques such as histology and cryosectioning demonstrate several limitations in studying adipose biology such as lengthy chemical processing steps, which can lead to tissue shrinkage and structure distortion3,4. Furthermore, these 2D techniques are insufficient to observe intercellular interactions exerted by different cell types, as the sections obtained are limited to smaller regions of the entire tissue3. Compared to conventional methods of fluorescent imaging, whole-mount staining does not require additional invasive steps, such as embedding, sectioning, and dehydration; thus, this avoids the problem of diminishing antibody specificity. As such, it is a simple and efficient method for imaging adipose tissue, with better preservation of adipocyte morphology&#160;and overall adipose tissue structure5. Therefore, whole-mount staining as a quick and inexpensive immunolabeling technique was established to preserve adipose tissue 3D architecture1,6,7,8.
However, despite the preservation of adipose tissue morphology with use of whole-mount staining, this technique is still unable to visualize inner structures beneath the lipid surface of the tissue. Several recent studies9,10&#160;have established tissue clearing techniques combined with whole-mount immunolabeling1,6&#160;to allow for comprehensive 3D visualization within adipose tissue. In particular, dense neural and vasculature networks were visualized in recent studies9,10,11,12&#160;with 3D volume imaging. Indeed, studying the neural and vascular plasticity of adipose tissue under different physiological conditions is essential to study its biology. Immunolabeling-enabled three-dimensional imaging of solvent-cleared organs (iDISCO+) tissue clearing is a process comprised of methanol pre-treatment, immunolabeling, and clearing of tissue opaqueness with organic chemical reagents dichloromethane (DCM) and dibenzyl ether (DBE)13,14. By making the adipose tissue entirely transparent, a more accurate representation of anatomy within the tissue such as blood vessels and neural fibers can be obtained9,10. IDISCO+ has advantages in that it is compatible with various antibodies and fluorescent reporters11,14, and it has demonstrated success in multiple organs and even embryoes14. However, its main limitation is a long incubation time, in which 18 to 20 days are needed to complete the entire experiment.
Another important application of whole-mount staining is the visualization of cell fate in combination with a lineage tracing system. Lineage tracing is the labelling of a specific gene/marker in a cell that can be passed on to all daughter cells and is conserved over time15. As such, it is a powerful tool that can be used to determine the fate of a cell&#39;s progeny15. Since the 1990s, the Cre-LoxP recombinant system has become a powerful approach for lineage tracing in living organisms15. When a mouse line that expresses Cre, a DNA recombinase enzyme, is crossed with another mouse line expressing a reporter that is adjacent to a loxP-STOP-loxP sequence, the reporter protein is expressed15.
For whole-mount staining, the use of fluorescent multicolor reporters is suitable for imaging of adipose tissue because it allows for minimal interference with intracellular activities of the adipocyte16. However, traditional reporters typically stain the cytoplasm, making it difficult to trace the lineage of white adipocytes, which have limited cytoplasmic content17. To overcome this problem, the use of membrane-bound fluorescent tdTomato/membrane eGFP (mT/mG) reporter marker is an ideal tool. Membrane-targeted tdTomato is expressed in Cre-negative cells18. Upon Cre excision, a switch to the expression of membrane-targeted eGFP occurs, making this reporter suitable for tracing the lineage of adipocyte progenitors17,18&#160;(Supplementary Figure 1).
The purpose of this paper is to provide a detailed protocol for whole-mount staining and show how it can be combined with other techniques to study the development and physiology of adipose tissue. Two examples of applications described in this protocol are its use with 1) multicolor reporter mouse lines to identify various origins of adipocytes and 2) tissue clearing to further visualize the neural arborization in white adipose tissue (WAT).

<table>
	<tbody>
		<tr>
			<td>LipidTox</td>
			<td>Life Technologies</td>
			<td>H34477</td>
			<td></td>
		</tr>
		<tr>
			<td>PECAM-1 primary antibody</td>
			<td>Millipore</td>
			<td>MAB1398Z(CH)</td>
			<td></td>
		</tr>
		<tr>
			<td>TH (tyrosine hydroxylase) primary antibody</td>
			<td>Millipore</td>
			<td>AB152, AB1542</td>
			<td></td>
		</tr>
		<tr>
			<td>DAPI stain</td>
			<td>BD Pharmingen</td>
			<td>564907</td>
			<td></td>
		</tr>
		<tr>
			<td>Nikon A1R confocal microscope</td>
			<td>Nikon</td>
			<td></td>
			<td>Confocal microscope</td>
		</tr>
		<tr>
			<td>Ultramicroscope I</td>
			<td>LaVision BioTec</td>
			<td></td>
			<td>Light sheet image fluorescent microscope</td>
		</tr>
		<tr>
			<td>Alexa Fluor secondary antibodies</td>
			<td>Jackson ImmunoResearch</td>
			<td></td>
			<td>Wavelengths 488, 594 and 647 used</td>
		</tr>
		<tr>
			<td>Purified Rat Anti-Mouse CD16/CD32</td>
			<td>BioSciences</td>
			<td>553141</td>
			<td></td>
		</tr>
		<tr>
			<td>Dichloromethane</td>
			<td>Sigma-Aldrich</td>
			<td>270997</td>
			<td></td>
		</tr>
		<tr>
			<td>Dibenzyl-ether</td>
			<td>Sigma-Aldrich</td>
			<td>33630</td>
			<td></td>
		</tr>
		<tr>
			<td>Methanol</td>
			<td>Fisher Chemical</td>
			<td>A452-1</td>
			<td></td>
		</tr>
		<tr>
			<td>30% Hydrogen Peroxide</td>
			<td>BIO BASIC CANADA INC</td>
			<td>HC4060</td>
			<td></td>
		</tr>
		<tr>
			<td>Dimethyl sulfoxide (DMSO)</td>
			<td>Sigma-Aldrich</td>
			<td>D2650</td>
			<td></td>
		</tr>
		<tr>
			<td>Glycine</td>
			<td>Sigma-Aldrich</td>
			<td>J7126</td>
			<td></td>
		</tr>
		<tr>
			<td>Heparin</td>
			<td>Sigma-Aldrich</td>
			<td>H3393</td>
			<td></td>
		</tr>
		<tr>
			<td>Lectin kit I, fluorescein labeled</td>
			<td>VECTOR LABORATORIES</td>
			<td>FLK-2100</td>
			<td></td>
		</tr>
		<tr>
			<td>F4/80</td>
			<td>Bio-Rad</td>
			<td>MCA497GA</td>
			<td></td>
		</tr>
		<tr>
			<td>VECTASHIELD Hard Set Mounting Medium with DAPI</td>
			<td>VECTOR LABORATORIES</td>
			<td>H-1500</td>
			<td></td>
		</tr>
		<tr>
			<td>Paraformaldehyde (PFA)</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Phosphate Buffer Saline (PBS)</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Triton-X</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Tween</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Animal serum (goat, donkey)</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
	</tbody>
</table>

visualization of 3d white adipose tissue structure using whole-mount staining

Adipose tissue is an important metabolic organ with high plasticity and is responsive to environmental stimuli and nutrient status. As such, various techniques have been developed to study the morphology and biology of adipose tissue. However, conventional visualization methods are limited to studying the tissue in 2D sections, failing to capture the 3D architecture of the whole organ. Here we present whole-mount staining, an immunohistochemistry method that preserves intact adipose tissue morphology with minimal processing steps. Hence, the structures of adipocytes and other cellular components are maintained without distortion, achieving the most representative 3D visualization of the tissue. In addition, whole-mount staining can be combined with lineage tracing methods to determine cell fate decisions. However, this technique has some limitations to providing accurate information regarding deeper parts of adipose tissue. To overcome this limitation, whole-mount staining can be further combined with tissue clearing techniques to remove the opaqueness of tissue and allow for complete visualization of entire adipose tissue anatomy using light-sheet fluorescent microscopy. Therefore, a higher resolution and more accurate representation of adipose tissue structures can be captured with the combination of these techniques.

Due to the fragility of adipose tissue, methods involving multiple processing steps and sectioning can lead to disfigurement of adipose tissue morphology3 (Figure 1A). However, whole-mount staining can preserve the morphology of adipocytes, ensuring accurate interpretation of results (Figure 1B).
Over-fixation of adipose tissue leads to fixative-induced autofluorescence. As shown in Figure ...

Watch this Scientific Journal Video about Visualization of 3D White Adipose Tissue Structure Using Whole-mount Staining at JoVE.com

Visualization of 3D White Adipose Tissue Structure Using Whole-mount Staining

The focus of the present study is to demonstrate the whole-mount immunostaining and visualization technique as an ideal method for 3D imaging of adipose tissue architecture and cellular component.

Adipose tissue is an important metabolic organ with high plasticity and is responsive to environmental stimuli and nutrient status. As such, various ...

Research

JoVE Journal

Biology

Method for Whole Mount Antibody Staining in Chick

The chick embryo is a valuable tool in the study of early embryonic development. Its transparency, accessibility and ease of manipulation, make it an ...

Zebrafish Whole Mount High-Resolution Double Fluorescent In Situ Hybridization

Zebrafish Whole Mount High-Resolution Double Fluorescent In Situ Hybridization

Whole mount in situ hybridization is one of the most widely used techniques in developmental biology.  Here, we present a high-resolution double ...

Using Whole Mount in situ Hybridization to Link Molecular and Organismal Biology

Using Whole Mount in situ Hybridization to Link Molecular and Organismal Biology

Whole mount in situ hybridization (WISH) is a common technique in molecular biology laboratories used to study gene expression through the localization of ...

Whole Mount in Situ Hybridization of E8.5 to E11.5 Mouse Embryos

Whole Mount in Situ Hybridization of E8.5 to E11.5 Mouse Embryos

Whole mount in situ hybridization is a very informative approach for defining gene expression patterns in embryos. The in situ hybridization procedures ...

Immunostaining of Whole-Mount Drosophila Testes for 3D Confocal Analysis of Large Spermatocytes

Immunostaining of Whole-Mount Drosophila Testes for 3D Confocal Analysis of Large Spermatocytes

Drosophila testes are a powerful model system for studying biological processes including stem cell biology, nuclear architecture, meiosis and sperm ...

Exploring Adipose Tissue Structure by Methylsalicylate Clearing and 3D Imaging

Obesity is a major worldwide public health issue that increases the risk to develop cardiovascular diseases, type-2 diabetes, and liver diseases. Obesity ...

Semi-Automated Isolation of Murine White Adipose Tissue

Semi-Automated Isolation of the Stromal Vascular Fraction from Murine White Adipose Tissue Using a Tissue Dissociator

Semi-Automated Isolation of the Stromal Vascular Fraction from Murine White Adipose Tissue Using a Tissue Dissociator (Video) | JoVE 

The in vitro study of white, brown, and beige adipocyte differentiation enables the investigation of cell-autonomous functions of adipocytes and their ...