Jm. Mackenzie et al., IMPROVED MEMBRANE PRESERVATION OF FLAVIVIRUS-INFECTED CELLS WITH CRYOSECTIONING, Journal of virological methods, 56(1), 1996, pp. 67-75
Citations number
17
Categorie Soggetti
Virology,"Biochemical Research Methods","Biothechnology & Applied Migrobiology
Ultra-cryomicrotomy and electron microscopy were used to investigate m
embranous structures in dengue virus-infected mammalian and insect cel
ls. The cryo-sectioned samples displayed ultrastructure comparable to
their resin-embedded counterparts with all previously identified virus
-induced structures being observed. Structures not previously identifi
ed were also found. In particular, membrane-bound packets of vesicles,
100-200 nm in diameter were seen distributed throughout areas of viru
s-induced membrane proliferation. These packets were clearly distinct
from virion arrays. Small smooth membrane vesicles, previously found t
o contain thread-like enclosures (M.L. Ng, J. Gen. Virol. 68 (1987) 57
7-582), were frequently observed to contain dense staining material, h
owever the exact nature of this material remains unclear. Virus-induce
d modification of golgi-like and/or ER membranes was also observed and
may represent early events in the generation of the smooth membrane v
esicles seen during infection. We suggest that cryosectioning is the m
ethod of choice to investigate membrane rearrangement induced by this
family of viruses and that a diamond knife and modified staining techn
iques, as utilised in this report, be employed to enhance morphology a
nd section preservation.