National Yang-Ming University, Institute of Biophotonics, and National Taiwan University Hospital, Department of Medical Research, Taipei 100, Taiwan.
J Biomed Opt. 2011 Jan-Feb;16(1):016012. doi: 10.1117/1.3528642.
Drosophila is one of the most valuable model organisms for studying genetics and developmental biology. The fat body in Drosophila, which is analogous to the liver and adipose tissue in human, stores lipids that act as an energy source during its development. At the early stages of metamorphosis, the fat body remodeling occurs involving the dissociation of the fat body into individual fat cells. Here we introduce a combination of coherent anti-Stokes Raman scattering (CARS) and two-photon excitation autofluorescence (TPE-F) microscopy to achieve label-free imaging of Drosophila in vivo at larval and pupal stages. The strong CARS signal from lipids allows direct imaging of the larval fat body and pupal fat cells. In addition, the use of TPE-F microscopy allows the observation of other internal organs in the larva and autofluorescent globules in fat cells. During the dissociation of the fat body, the findings of the degradation of lipid droplets and an increase in autofluorescent globules indicate the consumption of lipids and the recruitment of proteins in fat cells. Through in vivo imaging and direct monitoring, CARS microscopy may help elucidate how metamorphosis is regulated and study the lipid metabolism in Drosophila.
果蝇是研究遗传学和发育生物学最有价值的模式生物之一。果蝇的脂肪体类似于人类的肝脏和脂肪组织，储存脂质作为其发育过程中的能量来源。在变态早期，脂肪体发生重塑，脂肪体解离成单个脂肪细胞。在这里，我们介绍了相干反斯托克斯拉曼散射（CARS）和双光子激发自发荧光（TPE-F）显微镜的组合，以实现幼虫和蛹期活体果蝇的无标记成像。脂质的强 CARS 信号允许直接对幼虫脂肪体和蛹脂肪细胞进行成像。此外，TPE-F 显微镜的使用允许观察幼虫中的其他内部器官和脂肪细胞中的自发荧光球。在脂肪体解离过程中，脂质滴的降解和自发荧光球的增加的发现表明脂肪细胞中脂质的消耗和蛋白质的募集。通过活体成像和直接监测，CARS 显微镜可能有助于阐明变态是如何被调控的，并研究果蝇中的脂质代谢。
