Imaging of adipose tissue.
Adipose tissue is an endocrine organ that specializes in lipid metabolism and is distributed throughout the body in distinct white adipose tissue (WAT) and brown adipose tissue (BAT) depots. These tissues have opposing roles in lipid metabolism with WAT storing excessive caloric intake in the form of lipid, and BAT burning lipid through nonshivering thermogenesis. As accumulation of lipid in mature adipocytes of WAT leads to obesity and increased risk of comorbidity (Pi-Sunyer et al., 1998), detailed understanding of the mechanisms of BAT activation and WAT accumulation could produce therapeutic strategies for combatting metabolic pathologies. As morphological changes accompany alterations in adipose function, imaging of adipose tissue is one of the most important tools for understanding how adipose tissue mass fluctuates in response to various physiological contexts. Therefore, this chapter details several methods of processing and imaging adipose tissue, including bright-field colorimetric imaging of paraffin-sectioned adipose tissue with a detailed protocol for automated adipocyte size analysis; fluorescent imaging of paraffin and frozen-sectioned adipose tissue; and confocal fluorescent microscopy of whole mounted adipose tissue. We have also provided many example images showing results produced using each protocol, as well as commentary on the strengths and limitations of each approach.
Department of Molecular, Cell, and Developmental Biology, Yale University School of Medicine, New Haven, Connecticut, USA.
Ryan
R
Berry
Section of Comparative Medicine, Yale University School of Medicine, New Haven, Connecticut, USA.
Christopher D
CD
Church
Institute of Anatomy, University of Leipzig, Leipzig, Germany.
Martin T
MT
Gericke
Department of Cell Biology, Yale University School of Medicine, New Haven, Connecticut, USA.
Elise
E
Jeffery
Section of Comparative Medicine, Yale University School of Medicine, New Haven, Connecticut, USA.
Laura
L
Colman
Department of Molecular, Cell, and Developmental Biology, Yale University School of Medicine, New Haven, Connecticut, USA; Section of Comparative Medicine, Yale University School of Medicine, New Haven, Connecticut, USA; Yale Stem Cell Center, Yale University School of Medicine, New Haven, Connecticut, USA. Electronic address: matthew.rodeheffer@yale.edu.
Matthew S
MS
Rodeheffer
D002001:Adipose Tissue, Brown / Q000648:ultrastructure*; D052436:Adipose Tissue, White / Q000648:ultrastructure*; D050154:Adiposity / Q000235:genetics; D003952:Diagnostic Imaging; D006801:Humans; D050356:Lipid Metabolism; D018613:Microscopy, Confocal / Q000379:methods*; D009765:Obesity / Q000175:diagnosis / Q000235:genetics / Q000473:pathology*; D022722:Thermogenesis
Adipose; Cell profiler; Confocal; Frozen; Lineage tracing; Paraffin; Whole mount