open access

Vol 77, No 2 (2018)
ORIGINAL ARTICLES
Published online: 2017-08-31
Submitted: 2017-06-09
Accepted: 2017-07-24
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The influence of post-fixation on visualising vimentin in the retina using immunofluorescence method

B. Baykal, C. Korkmaz, N. Kocabiyik, O.M. Ceylan
DOI: 10.5603/FM.a2017.0082
·
Pubmed: 28868606
·
Folia Morphol 2018;77(2):246-252.

open access

Vol 77, No 2 (2018)
ORIGINAL ARTICLES
Published online: 2017-08-31
Submitted: 2017-06-09
Accepted: 2017-07-24

Abstract

Background: Post-fixation of sections is especially required for cryostat sections of fresh frozen tissues. Vimentin is an intermediate filament in both fibrillary and non-fibrillary form, expressed in Müller’s cells and astrocytes of the retina. Our aim was to determine the best post-fixation method for visualising vimentin in archival mouse eyes.

Materials and methods: We used an archival mouse eye, slightly pre-fixed with paraformaldehyde and stored at –80°C for 4 years. We tried three fixatives (pa­raformaldehyde [PFA], alcohol/acetic acid [AAA] and methanol) for post-fixation of eye sections.

Results: We showed that post-fixation alters the labelling properties of vimentin expressed in the retina. In the sections with no post-fixation, vimentin positivity was observed in and around the nuclei in non-fibrillary form. In PFA post-fixed sections, the vimentin in the retina was not observed as fibrils. Positivity was observed in the nuclei and in perinuclear regions of the cells. In AAA post-fixed sections, positive labelling was observed around the nuclei as fibrils. In methanol post-fixed sections, labelling was observed around the nuclei as fibrils.

Conclusions: We conclude that post-fixation with AAA is more convenient for immunofluorescent labelling of vimentin in the retina for slightly PFA pre-fixed and long-term stored retina. (Folia Morphol 2018; 77, 2: 246–252)

Abstract

Background: Post-fixation of sections is especially required for cryostat sections of fresh frozen tissues. Vimentin is an intermediate filament in both fibrillary and non-fibrillary form, expressed in Müller’s cells and astrocytes of the retina. Our aim was to determine the best post-fixation method for visualising vimentin in archival mouse eyes.

Materials and methods: We used an archival mouse eye, slightly pre-fixed with paraformaldehyde and stored at –80°C for 4 years. We tried three fixatives (pa­raformaldehyde [PFA], alcohol/acetic acid [AAA] and methanol) for post-fixation of eye sections.

Results: We showed that post-fixation alters the labelling properties of vimentin expressed in the retina. In the sections with no post-fixation, vimentin positivity was observed in and around the nuclei in non-fibrillary form. In PFA post-fixed sections, the vimentin in the retina was not observed as fibrils. Positivity was observed in the nuclei and in perinuclear regions of the cells. In AAA post-fixed sections, positive labelling was observed around the nuclei as fibrils. In methanol post-fixed sections, labelling was observed around the nuclei as fibrils.

Conclusions: We conclude that post-fixation with AAA is more convenient for immunofluorescent labelling of vimentin in the retina for slightly PFA pre-fixed and long-term stored retina. (Folia Morphol 2018; 77, 2: 246–252)

Get Citation

Keywords

vimentin, post-fixation, paraformaldehyde, alcohol/acetic acid, methanol

About this article
Title

The influence of post-fixation on visualising vimentin in the retina using immunofluorescence method

Journal

Folia Morphologica

Issue

Vol 77, No 2 (2018)

Pages

246-252

Published online

2017-08-31

DOI

10.5603/FM.a2017.0082

Pubmed

28868606

Bibliographic record

Folia Morphol 2018;77(2):246-252.

Keywords

vimentin
post-fixation
paraformaldehyde
alcohol/acetic acid
methanol

Authors

B. Baykal
C. Korkmaz
N. Kocabiyik
O.M. Ceylan

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