Rapid tool for cell nanoarchitecture integrity assessment.
J Struct Biol
; 213(4): 107801, 2021 12.
Article
em En
| MEDLINE
| ID: mdl-34582983
ABSTRACT
With the rapid increase and accessibility of high-resolution imaging technologies of cells, the interpretation of results relies more and more on the assumption that the three-dimensional integrity of the surrounding cellular landscape is not compromised by the experimental setup. However, the only available technology for directly probing the structural integrity of whole-cell preparations at the nanoscale is electron cryo-tomography, which is time-consuming, costly, and complex. We devised an accessible, inexpensive and reliable screening assay to quickly report on the compatibility of experimental protocols with preserving the structural integrity of whole-cell preparations at the nanoscale. Our Rapid Cell Integrity Assessment (RCIA) assay is executed at room temperature and relies solely on light microscopy imaging. Using cellular electron cryo-tomography as a benchmark, we verify that RCIA accurately unveils the adverse impact of reagents and/or protocols such as those used for virus inactivation or to arrest dynamic processes on the cellular nanoarchitecture.
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Texto completo:
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Base de dados:
MEDLINE
Assunto principal:
Microscopia Crioeletrônica
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Imageamento Tridimensional
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Nanoestruturas
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Células Eucarióticas
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Tomografia com Microscopia Eletrônica
Limite:
Animals
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Humans
Idioma:
En
Ano de publicação:
2021
Tipo de documento:
Article