Methods for making and observing model lipid droplets.
Cell Rep Methods
; 4(5): 100774, 2024 May 20.
Article
en En
| MEDLINE
| ID: mdl-38749444
ABSTRACT
We present methods for making and testing the membrane biophysics of model lipid droplets (LDs). Methods are described for imaging LDs ranging in size from 0.1 to 40 µm in diameter with high-resolution microscopy and spectroscopy. With known LD compositions, membrane binding, sorting, diffusion, and tension were measured via fluorescence correlation spectroscopy (FCS), fluorescence recovery after photobleaching (FRAP), fluorescence lifetime imaging microscopy (FLIM), atomic force microscopy (AFM), and imaging flow cytometry. Additionally, a custom, small-volume pendant droplet tensiometer is described and used to measure the association of phospholipids to the LD surface. These complementary, cross-validating methods of measuring LD membrane behavior reveal the interplay of biophysical processes on lipid droplet monolayers.
Palabras clave
AFM; CP: Biotechnology; CP: Cell biology; FCS; FLIM; FRAP; atomic force microscopy; diffusion; fluorescence correlation spectroscopy; fluorescence lifetime imaging microscopy; fluorescence microscopy; fluorescence recovery after photobleaching; model lipid droplets; pendant droplet tensiometry; phospholipid monolayers; sorting; tension
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1
Base de datos:
MEDLINE
Asunto principal:
Gotas Lipídicas
Idioma:
En
Revista:
Cell Rep Methods
Año:
2024
Tipo del documento:
Article