Quantitative Fluorescence In Situ Hybridization (FISH) and Immunofluorescence (IF) of Specific Gene Products in KSHV-Infected CellsTenaya K. Vallery 1, Joan A. Steitz 2
1Norfolk Academy, 2Department of Molecular Biophysics and Biochemistry, Howard Hughes Medical Institute, Yale School of Medicine
We describe a protocol utilizing fluorescence in situ hybridization (FISH) to visualize multiple herpesviral RNAs within lytically infected human cells, either in suspension or adherent. This protocol includes quantification of fluorescence producing a nucleocytoplasmic ratio and can be extended for simultaneous visualization of host and viral proteins with immunofluorescence (IF).
