Challenges of Using Expansion Microscopy for Super-resolved Imaging of Cellular Organelles.
STED microscopy
bioorganic chemistry
cell organelles
expansion microscopy
peptides
Journal
Chembiochem : a European journal of chemical biology
ISSN: 1439-7633
Titre abrégé: Chembiochem
Pays: Germany
ID NLM: 100937360
Informations de publication
Date de publication:
15 02 2021
15 02 2021
Historique:
received:
13
08
2020
revised:
07
10
2020
pubmed:
14
10
2020
medline:
28
9
2021
entrez:
13
10
2020
Statut:
ppublish
Résumé
Expansion microscopy (ExM) has been successfully used to improve the spatial resolution when imaging tissues by optical microscopy. In ExM, proteins of a fixed sample are crosslinked to a swellable acrylamide gel, which expands when incubated in water. Therefore, ExM allows enlarged subcellular structures to be resolved that would otherwise be hidden to standard confocal microscopy. Herein, we aim to validate ExM for the study of peroxisomes, mitochondria, nuclei and the plasma membrane. Upon comparison of the expansion factors of these cellular compartments in HEK293 cells within the same gel, we found significant differences, of a factor of above 2, in expansion factors. For peroxisomes, the expansion factor differed even between peroxisomal membrane and matrix marker; this underlines the need for a thorough validation of expansion factors of this powerful technique. We further give an overview of possible quantification methods for the determination of expansion factors of intracellular organelles, and we highlight some potentials and challenges.
Identifiants
pubmed: 33049107
doi: 10.1002/cbic.202000571
pmc: PMC7894168
doi:
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Research Support, U.S. Gov't, Non-P.H.S.
Langues
eng
Sous-ensembles de citation
IM
Pagination
686-693Subventions
Organisme : Medical Research Council
ID : MR/S005382/1
Pays : United Kingdom
Organisme : John Fell Fund
Organisme : MRC
ID : G0902418
Organisme : Medical Research Council
ID : MC_UU_00008/9
Pays : United Kingdom
Organisme : Wolfson Foundation
Organisme : EPA Cephalosporin Fund
Organisme : Medical Research Council
ID : MC_UU_12010/9
Pays : United Kingdom
Organisme : German Research Foundation (DFG)
ID : 322325142
Organisme : Wolfson Imaging Centre, Oxford
Organisme : MRC
ID : MC_UU_12010
Organisme : Wellcome Trust
ID : 091911
Pays : United Kingdom
Organisme : MRC
ID : MC_UU_12025
Organisme : Wellcome Trust
ID : 104924/14/Z/14
Pays : United Kingdom
Organisme : MRC/BBSRC/EPSRC
ID : MR/K01577X/1
Organisme : Wellcome Trust
Pays : United Kingdom
Organisme : Wellcome Trust
ID : 104924/14/Z/14
Pays : United Kingdom
Informations de copyright
© 2020 The Authors. ChemBioChem published by Wiley-VCH GmbH.
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