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HERO ID
3859472
Reference Type
Journal Article
Title
PARAFORMALDEHYDE FIXATION OF NEUTROPHILS FOR IMMUNOLABELING OF GRANULE ANTIGENS IN CRYOULTRASECTIONS
Author(s)
Elliot, E; Dennison, C; Fortgens, PH; Travis, J
Year
1995
Is Peer Reviewed?
Yes
Journal
Journal of Histochemistry and Cytochemistry
ISSN:
0022-1554
EISSN:
1551-5044
Volume
43
Issue
10
Page Numbers
1019-1025
Web of Science Id
WOS:A1995RW32600006
Abstract
Paraformaldehyde (PFA) fixation was optimized to facilitate the immobilization and labeling of multiple granule antigens, using short fixation regimens and cryoultramicrotomy of unembedded neutrophils (PMNs). In the optimal protocol, extraction of azurophil granule antigens (especially of the abundant elastase) was obviated by manipulating the polymeric state of PFA, and hence its rate of cross-linking, by altering its concentration and pH in a multistep process, Primary fixation conditions used (4% PFA, pH 8.0, 5 min) favor fixative penetration and rapid cross-linking, Stable crosslinking of the antigen was achieved in a secondary fixation step using conditions that favor larger, more cross-linking polymeric forms of PFA (8% PFA, pH 7.2, 15 min), Immobilization of granule antigens was enhanced by flotation of cut sections on fixative (8% PFA, pH 8.0) before labeling and by using post-labeling fixation with 1% glutaraldehyde. The optimized protocol facilitated immobilization and immunolabeling of elastase, myeloperoxidase, lactoferrin, and cathepsin D in highly hydrated, unembedded PMNs.
Keywords
PARAFORMALDEHYDE FIXATION; CRYOULTRAMICROTOMY; ELASTASE; CATHEPSIN D; MYELOPEROXIDASE; LACTOFERRIN; NEUTROPHILS
Tags
PFAS
•
FtOH 8:2
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WOS
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Excluded/Not on Topic
•
^Per- and Polyfluoroalkyl Substances (PFAS)
FtOH 8:2 (678-39-7)
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WOS
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