Intracellular flow cytometric lipid analysis - a multiparametric system to assess distinct lipid classes in live cells.
J Cell Sci
; 135(5)2022 03 01.
Article
em En
| MEDLINE
| ID: mdl-34114626
ABSTRACT
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.
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Texto completo:
1
Base de dados:
MEDLINE
Assunto principal:
Colesterol
/
Ésteres do Colesterol
Limite:
Animals
Idioma:
En
Revista:
J Cell Sci
Ano de publicação:
2022
Tipo de documento:
Article
País de afiliação:
Austrália