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Reducing Frost during Cryoimaging Using a Hygroscopic Ice Frame

[Image: see text] Cryomicroscopy is commonly hampered by frost accumulation, reducing the visual clarity of the specimen. Pulling a vacuum or purging with nitrogen gas can greatly reduce the sample chamber’s humidity, but at cryogenic temperatures, even minute concentrations of water vapor can still...

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Detalles Bibliográficos
Autores principales: Lowery, Adam W., Ambi, Ashwin, Miller, Lisa M., Boreyko, Jonathan B.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: American Chemical Society 2022
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9730467/
https://www.ncbi.nlm.nih.gov/pubmed/36506191
http://dx.doi.org/10.1021/acsomega.2c03083
Descripción
Sumario:[Image: see text] Cryomicroscopy is commonly hampered by frost accumulation, reducing the visual clarity of the specimen. Pulling a vacuum or purging with nitrogen gas can greatly reduce the sample chamber’s humidity, but at cryogenic temperatures, even minute concentrations of water vapor can still result in frost deposition. Here, a hygroscopic ice frame was created around the specimen to suppress frost growth during cryomicroscopy. Specifically, fluorescently tagged rat brain vessels were frozen on a silicon nitride window with an ice frame, and the luminescence of the fluorescent tag was improved by a factor of 6 compared to a similar specimen in only a nitrogen purge environment. These findings suggest that the simple implementation of a hygroscopic ice frame surrounding the specimen can substantially improve the visual clarity for cryomicroscopy, beyond that of a vacuum or nitrogen purge system.