a comparative study of fat storage quantitation in nematode caenorhabditis elegans using label and label-free methods脂肪储存定量的比较研究线虫线虫使用标签和label-free方法.pdfVIP

A Comparative Study of Fat Storage Quantitation in Nematode Caenorhabditis elegans Using Label and Label-Free Methods 1. 2. 1 1 2,3,4 Kelvin Yen , Thuc T. Le , Ankita Bansal , Sri Devi Narasimhan , Ji-Xin Cheng *, Heidi A. Tissenbaum1,5* 1 Program in Gene Function and Expression, University of Massachusetts Medical School, Worcester, Massachusetts, United States of America, 2 Weldon School of Biomedical Engineering, Purdue University, West Lafayette, Indiana, United States of America, 3 Purdue Cancer Center, Purdue University, West Lafayette, Indiana, United States of America, 4 Department of Chemistry, Purdue University, West Lafayette, Indiana, United States of America, 5 Program in Molecular Medicine, University of Massachusetts Medical School, Worcester, Massachusetts, United States of America Abstract The nematode Caenorhabditis elegans has been employed as a model organism to study human obesity due to the conservation of the pathways that regulate energy metabolism. To assay for fat storage in C. elegans, a number of fat- soluble dyes have been employed including BODIPY, Nile Red, Oil Red O, and Sudan Black. However, dye-labeled assays produce results that often do not correlate with fat stores in C. elegans. An alternative label-free approach to analyze fat storage in C. elegans has recently been described with coherent anti-Stokes Raman scattering (CARS) microscopy. Here, we compare the performance of CARS microscopy with standard dye-labeled techniques and biochemical quantification to analyze fat storage in wild type C. elegans and with genetic mutations in the insulin/IGF-1 signaling pathway including the genes daf-2 (insulin/IGF-1