Berry Ryan, Church Christopher D, Gericke Martin T, Jeffery Elise, Colman Laura, Rodeheffer Matthew S
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.
Methods Enzymol. 2014;537:47-73. doi: 10.1016/B978-0-12-411619-1.00004-5.
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.
脂肪组织是一个专门负责脂质代谢的内分泌器官，以不同的白色脂肪组织（WAT）和棕色脂肪组织（BAT）储存库的形式分布于全身。这些组织在脂质代谢中具有相反的作用，白色脂肪组织以脂质的形式储存过量的热量摄入，而棕色脂肪组织则通过非寒战产热消耗脂质。由于白色脂肪组织成熟脂肪细胞中脂质的积累会导致肥胖和合并症风险增加（Pi-Sunyer等人，1998年），详细了解棕色脂肪组织激活和白色脂肪组织积累的机制可能会产生对抗代谢疾病的治疗策略。由于脂肪功能的改变伴随着形态学变化，脂肪组织成像对于理解脂肪组织质量如何响应各种生理环境而波动是最重要的工具之一。因此，本章详细介绍了几种处理和成像脂肪组织的方法，包括石蜡切片脂肪组织的明场比色成像以及用于自动分析脂肪细胞大小的详细方案；石蜡和冰冻切片脂肪组织的荧光成像；以及全装脂肪组织的共聚焦荧光显微镜检查。我们还提供了许多示例图像，展示了使用每种方案产生的结果，以及对每种方法的优缺点的评论。
