Lowery, Adam W.Ambi, AshwinMiller, Lisa M.Boreyko, Jonathan B.2023-04-182023-04-182022-12http://hdl.handle.net/10919/114542Cryomicroscopy 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.application/pdfenCreative Commons Attribution-NonCommercial-NoDerivatives 4.0 InternationalX-ray-fluorescencecryoelectron microscopyelectron-microscopyvapor-pressurescryomicroscopylightautofluorescencewaterArticle - Refereedhttps://doi.org/10.1021/acsomega.2c0308375136506191