Harris Lydia-Ann L S, Skinner James R, Wolins Nathan E
Center for Human Nutrition, Washington University School of Medicine, St. Louis, Missouri, USA.
Methods Cell Biol. 2013;116:213-26. doi: 10.1016/B978-0-12-408051-5.00011-5.
Intracellular fat droplets are large and have a distinct morphology, which makes their imaging at the light level simple and informative. We detail how to image the fat droplet core by metabolic labeling with fluorescent fatty acids or lipophilic fluorochromes. Further, we describe the use of indirect immunostaining to image fat droplet proteins and fat cores in the same field. We also address the use of appropriate controls for determining signal specificity and other practical considerations for optimizing image quality.
细胞内的脂肪滴很大，且具有独特的形态，这使得在光学水平上对其成像既简单又能提供丰富信息。我们详细介绍了如何通过用荧光脂肪酸或亲脂性荧光染料进行代谢标记来对脂肪滴核心进行成像。此外，我们描述了如何使用间接免疫染色在同一视野中对脂肪滴蛋白和脂肪核心进行成像。我们还讨论了使用适当的对照来确定信号特异性以及优化图像质量的其他实际考虑因素。
