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Updated: May 5, 2026
Adipo-Clear: A Tissue Clearing Method for Three-Dimensional Imaging of Adipose Tissue
Published on: July 28, 2018
Application of immunocytochemistry and immunofluorescence techniques to adipose tissue and cell cultures
1Light Microscopy Core Facility, National Heart, Lung and Blood Institute, National Institutes of Health, Bethesda, MD, USA.
Insights
Confocal microscopy enables the study of whole white adipose cells, overcoming challenges posed by their structure and buoyancy. This technique allows detailed observation of insulin-regulated protein trafficking within these crucial energy-storing cells.
Area of Science:
- Cell Biology
- Physiology
- Microscopy
Background:
- White adipose cells store triglycerides, presenting microscopy challenges due to their large lipid droplet and buoyancy.
- Studying intact adipocytes is vital for understanding hormone-mediated responses, as cultured models may not fully replicate physiological significance.
Purpose of the Study:
- To detail the application of confocal microscopy and image reconstruction for studying whole adipose cells.
- To demonstrate imaging of whole-mount living adipose tissue for in situ cellular analysis.
Main Methods:
- Utilizing confocal microscopy for optical sectioning of whole adipose cells.
- Employing computer-assisted image reconstruction to analyze cellular structures.
- Developing techniques for imaging whole-mount preparations of living adipose tissue.
Main Results:
- Successfully applied confocal microscopy to investigate insulin-regulated protein trafficking in whole adipose cells.
- Demonstrated the feasibility of imaging intact adipose tissue, preserving cellular interactions.
- Overcame challenges associated with the unique morphology and buoyancy of adipocytes for microscopy.
Conclusions:
- Confocal microscopy offers an effective morphological approach for studying adipose cell physiology and pathophysiology.
- In situ imaging of adipose tissue provides new avenues for understanding adipocyte function within its native environment.
- Advanced microscopy techniques are crucial for detailed analysis of adipocyte behavior and responses.
Abstract:
When isolated from tissue, white adipose cells are round, and their interior is filled with a large (80-120 microm) droplet of stored triglyceride, leaving a thin (1-2-microm) layer of cytoplasm between the lipid droplet and the plasma membrane. Their three-dimensional architecture, together with the fact that these cells ordinarily float in medium, have created major challenges when one attempts to perform microscopy techniques with these cells. Adipocytes serve as the principal energy reservoir in the body, and it is essential to overcome these difficulties to be able to study hormone-mediated responses in real adipose cells, which convey physiological significance that cannot be readily duplicated by the use of cultured model adipocytes. This chapter focuses on the use of confocal microscopy optical sectioning and computer-assisted image reconstruction in the whole adipose cell in the study of insulin-regulated protein trafficking. In addition, we illustrate the possibility to image whole-mount preparations of living adipose tissue, opening new ways to probe adipose cells in situ without disrupting their cellular interactions within living adipose tissue. Confocal microscopy constitutes an effective morphological approach to investigating adipose cell physiology and pathophysiology.
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