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CryoFISH: fluorescence in situ hybridization on ultrathin cryosections

Item Type:Article
Title:CryoFISH: fluorescence in situ hybridization on ultrathin cryosections
Creators Name:Xie, S.Q. and Lavitas, L.M. and Pombo, A.
Abstract:The visualization of cellular structures and components has become an invaluable tool in biological and medical sciences. Imaging subcellular compartments and single molecules within a cell has prompted the development of a wide range of sample preparation techniques as well as various microscope devices to obtain images with increased spatial resolution. Here, we present cryoFISH, a method for fluorescence in situ hybridization (FISH) on thin (~150 nm thick) cryosections from sucrose-embedded fixed cells or tissues. CryoFISH can be used in combination with immunodetection (IF) of other cellular components. The main advantages of cryoFISH and cryoIF over whole-cell labeling methods are increased spatial resolution with confocal microscopy, greater sensitivity of detection due to increased probe accessibility, and better image contrast. CryoFISH and cryoIF methods typically used on samples fixed in conditions that preserve ultrastructure, are compatible with the labeling of cells in their tissue context and are ideal for correlative studies that compare fluorescence with electron microscopy.
Keywords:Imaging, Immunofluorescence, Fluorescence In-Situ Hybridization, Interchromosomal Interactions, Tokuyasu Cryosections, RNA Polymerase II, Animals, Mice
Source:Methods in Molecular Biology
Publisher:Springer / Humana Press
Page Range:219-230
Official Publication:https://doi.org/10.1007/978-1-60761-789-1_15
PubMed:View item in PubMed

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