Journal
JOURNAL OF CELL SCIENCE
Volume 135, Issue 5, Pages -Publisher
COMPANY BIOLOGISTS LTD
DOI: 10.1242/jcs.258322
Keywords
Flow cytometry; Imaging; Lipids; Lipophilic dyes; Nile Blue; Nile Red
Categories
Funding
- NWG Macintosh Memorial Award
- Macintosh Scholarship
- Kickstarter Award from The University of Sydney Nano Institute
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This article describes a simple and inexpensive method to determine lipid content in cells, and highlights the differences in lipid biology between cell types using imaging and flow cytometry.
The lipid content of mammalian cells varies greatly between cell type. Current methods for analysing lipid components of cells are technically challenging and destructive. Here, we report a facile, inexpensive method to identify lipid content - intracellular flow cytometric lipid analysis (IFCLA). Distinct lipid classes can be distinguished by Nile Blue fluorescence, Nile Red fluorescence or violet autofluorescence. Nile Blue is fluorescent in the presence of unsaturated fatty acids with a carbon chain length greater than 16. Cis-configured fatty acids induce greater Nile Blue fluorescence than their trans-configured counterparts. In contrast, Nile Red exhibits greatest fluorescence in the presence of cholesterol, cholesteryl esters, some triglycerides and phospholipids. Multiparametric spanning-tree progression analysis for density-normalized events (SPADE) analysis of hepatic cellular lipid distribution, including vitamin A autofluorescence, is presented. This flow cytometric system allows for the rapid, inexpensive and non-destructive identification of lipid content, and highlights the differences in lipid biology between cell types by imaging and flow cytometry. This article has an associated First Person interview with the first author of the paper.
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