Single-molecule RNA FISH (CAT#: STEM-MB-1176-WXH)

Introduction

Single-molecule RNA FISH, also known as Stellaris® RNA FISH or smFISH, is a method of detecting and quantifying mRNA and other long RNA molecules in a thin layer of tissue sample.
Single-molecule RNA FISH assays can be performed in simplex or multiplex, and can be used as a follow-up experiment to quantitative PCR, or imaged simultaneously with a fluorescent antibody assay.

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Principle Applications Procedure Materials Notes Related Products

Principle

Targets can be reliably imaged through the application of multiple short singly labeled oligonucleotide probes. The binding of up to 48 fluorescent labeled oligos to a single molecule of mRNA provides sufficient fluorescence to accurately detect and localize each target mRNA in a wide-field fluorescent microscopy image.

Applications

Detecting and quantifying mRNA and other long RNA molecules in a thin layer of tissue sample.
The technology has potential applications in cancer diagnosis, neuroscience, gene expression analysis, and companion diagnostics.

Procedure

1. Design and order RNA FISH probes
2. Prepare cultures
3. Fix and spheroplast yeast cells
4. Cells are hybridized with the fluorescent oligonucleotides, washed, and mounted onto coverslips.
5. Imaging

Materials

• Flow cytometer
• Fluorescence microscopy

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