Super-Resolution Imaging of the A- and B-Type Lamin Networks: A Comparative Study of Different Fluorescence Labeling Procedures.
STED microscopy
antibodies
colocalization
confocal scanning laser microscopy
lamin layer thickness
laminopathy
nuclear lamins
resolution
transfection
Journal
International journal of molecular sciences
ISSN: 1422-0067
Titre abrégé: Int J Mol Sci
Pays: Switzerland
ID NLM: 101092791
Informations de publication
Date de publication:
22 Sep 2021
22 Sep 2021
Historique:
received:
22
06
2021
revised:
13
09
2021
accepted:
14
09
2021
entrez:
13
10
2021
pubmed:
14
10
2021
medline:
28
10
2021
Statut:
epublish
Résumé
A- and B-type lamins are type V intermediate filament proteins. Mutations in the genes encoding these lamins cause rare diseases, collectively called laminopathies. A fraction of the cells obtained from laminopathy patients show aberrations in the localization of each lamin subtype, which may represent only the minority of the lamina disorganization. To get a better insight into more delicate and more abundant lamina abnormalities, the lamin network can be studied using super-resolution microscopy. We compared confocal scanning laser microscopy and stimulated emission depletion (STED) microscopy in combination with different fluorescence labeling approaches for the study of the lamin network. We demonstrate the suitability of an immunofluorescence staining approach when using STED microscopy, by determining the lamin layer thickness and the degree of lamin A and B1 colocalization as detected in fixed fibroblasts (co-)stained with lamin antibodies or (co-)transfected with EGFP/YFP lamin constructs. This revealed that immunofluorescence staining of cells does not lead to consequent changes in the detected lamin layer thickness, nor does it influence the degree of colocalization of lamin A and B1, when compared to the transfection approach. Studying laminopathy patient dermal fibroblasts (
Identifiants
pubmed: 34638534
pii: ijms221910194
doi: 10.3390/ijms221910194
pmc: PMC8508656
pii:
doi:
Substances chimiques
Intermediate Filament Proteins
0
Lamin Type A
0
Lamin Type B
0
Types de publication
Journal Article
Langues
eng
Sous-ensembles de citation
IM
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