A rapid method for staining large chylomicrons.
In this report, we present a rapid method for producing high-quality micrographs suitable for determining the size distributions of particles in concentrated samples of postprandial chylomicrons and chylomicron remnants. The procedure consists of mixing particles with osmium tetroxide in water to stabilize the lipids of the particles. These fixed and positively stained particles are then negatively stained with phosphotungstate in the presence of dilute sucrose. This dual staining procedure prevents the fusion and clustering of chylomicrons during processing for electron microscopy and is effective with particles of different lipid compositions. In addition, this procedure is simple and rapid, adding only one mixing step and 5 min to the preparation time required for conventional negative stains.
Anderson
L J
LJ
Gladstone Foundation Laboratories for Cardiovascular Disease, Cardiovascular Research Institute, San Francisco, CA.
Boyles
J K
JK
Hussain
M M
MM
D000818:Animals; D002914:Chylomicrons / Q000032:analysis*; D004285:Dogs; D004492:Edetic Acid; D008196:Lymph / Q000032:analysis; D008854:Microscopy, Electron; D009993:Osmium Tetroxide; D010316:Particle Size; D010772:Phosphotungstic Acid; D013194:Staining and Labeling / Q000379:methods*; D013395:Sucrose; D013897:Thoracic Duct / Q000032:analysis
D002914:Chylomicrons; D010772:Phosphotungstic Acid; D013395:Sucrose; D004492:Edetic Acid; D009993:Osmium Tetroxide