Method for localization of sialic acid on cell surface and cell interior in peripheral blood: a pilot study.
This article presents a method for identification and localization of cell surface and intracellular sialoglycoconjugates of peripheral blood cells. To reveal cell surface conjugates, a sample of peripheral blood was incubated with lectin after centrifugation and rinsing. For intracellular localization in leukocytes, RBCs were lysed and the membranes were permeabilized prior to cytochemical reaction. Fluorescein isothiocyanate conjugated lectins were used for visualization in fluorescence microscope. All lectins bound specifically to the surface of erythrocytes. Confocal microscopy showed surface and intracellular labeling of permeabilized leukocytes. A part of the signal in eosinophils originated from binding of anionic fluorophore to cationic granular proteins.
Pönniö
Maritta
M
Department of Clinical Neuroscience, Karolinska Hospital, Stockholm, Sweden. maritta.ponnio@cns.ki.se
Soukka
Jori
J
Sillanaukee
Pekka
P
Franck
Johan
J
Seveus
Lahja
L
D000328:Adult; D004912:Erythrocytes / Q000737:chemistry*; D006801:Humans; D037102:Lectins / Q000378:metabolism; D007962:Leukocytes / Q000737:chemistry*; D018613:Microscopy, Confocal; D008856:Microscopy, Fluorescence; D019158:N-Acetylneuraminic Acid / Q000097:blood*; D010865:Pilot Projects
D037102:Lectins; D019158:N-Acetylneuraminic Acid