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Dynamic study of intramembranous particles in human fresh erythrocytes using an “in vitro cryotechnique”
Author(s) -
Terada Nobuo,
Ohno Nobuhiko,
Fujii Yasuhisa,
Baba Takeshi,
Ohno Shinichi
Publication year - 2006
Publication title -
microscopy research and technique
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.536
H-Index - 118
eISSN - 1097-0029
pISSN - 1059-910X
DOI - 10.1002/jemt.20315
Subject(s) - intramembranous ossification , membrane , biophysics , chemistry , anatomy , biology , biochemistry
For analyses of dynamic ultrastructures of erythrocyte intramembranous particles (IMPs) in situ, a quick‐freezing method was used to stabilize the flow behavior of erythrocytes embedded in vitreous ice. Fresh human blood was jetted at various pressures through artificial tubes, in which the flowing erythrocytes were elongated from biconcave discoid shapes to elliptical ones, and quickly frozen in liquid isopentane–propane cryogen (−193°C). They were freeze‐fractured using a scalpel in liquid nitrogen, and routinely prepared for replica membranes. Many IMPs were observed on the protoplasmic freeze‐fracture face (P‐face) of the erythrocyte membranes. Some control erythrocytes under nonflowing or stationary conditions showed IMPs with their random distribution. However, other jetted erythrocytes under flowing conditions showed variously sized IMPs with much closer distribution. They were also arranged into parallel rows in some parts, and aggregated together. This quick‐freezing method enabled for the first time the visualization of time‐dependent topology and the molecular alteration of IMPs in dynamically flowing erythrocytes. Microsc. Res. Tech. 69: 291–295, 2006 © 2006 Wiley‐Liss, Inc.