Depiction of the RNA FISH scheme and demonstration of rapid hybridization.

A. Schematic of the single molecule RNA FISH method, in which we use dozens of short fluorescently labeled oligonucleotides that all target the same RNA molecule. B. Image showing RNA FISH targeting mRNA from the TBCB gene under standard overnight hybridization conditions (formaldehyde fixation). Each spot is a single mRNA molecule. C. Image showing RNA FISH signals from an attempt at rapid hybridization (5 minutes) with a high concentration of probe but with formaldehyde fixation. D., E. Traditional overnight hybridization and Turbo RNA FISH hybridization using methanol-fixed cells. All images are maximum projections of a stack of optical sections encompassing the three-dimensional volume of the cell. DAPI (nuclear stain) is in purple.