The influence of post-fixation on visualising vimentin in the retina using immunofluorescence method
Folia Morphologica (Poland), cilt.77, sa.2, ss.246-252, 2018 (SCI-Expanded, Scopus)
- Yayın Türü: Makale / Tam Makale
- Cilt numarası: 77 Sayı: 2
- Basım Tarihi: 2018
- Doi Numarası: 10.5603/fm.a2017.0082
- Dergi Adı: Folia Morphologica (Poland)
- Derginin Tarandığı İndeksler: Science Citation Index Expanded (SCI-EXPANDED), Scopus
- Sayfa Sayıları: ss.246-252
- Anahtar Kelimeler: vimentin, post-fixation, paraformaldehyde, alcohol/acetic acid, methanol
- Sağlık Bilimleri Üniversitesi Adresli: Evet
Özet
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.