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RNA Imaging with Multiplexed Error-Robust Fluorescence In Situ Hybridization (MERFISH)
1Howard Hughes Medical Institute, Harvard University, Cambridge, MA, United States.
Methods in Enzymology
|June 1, 2016
Summary
Multiplexed error-robust fluorescence in situ hybridization (MERFISH) enables simultaneous measurement of thousands of RNA species in single cells. This massively parallel approach provides unprecedented insights into cellular and tissue behaviors.
Area of Science:
- Molecular Biology
- Genomics
- Cell Biology
Background:
- Understanding cellular and tissue behaviors requires quantitative measurement of RNA copy number and spatial distribution.
- Single-molecule fluorescence in situ hybridization (smFISH) offers spatial and copy number RNA data but is limited in the number of detectable RNA species.
Purpose of the Study:
- To introduce multiplexed error-robust fluorescence in situ hybridization (MERFISH), a massively parallelized smFISH technique.
- To enable simultaneous imaging and identification of hundreds to thousands of RNA species in single cells with high accuracy.
Main Methods:
- MERFISH utilizes a massively parallelized approach to smFISH.
- The method involves specific probe design, advanced data collection, and robust data analysis protocols.
Main Results:
- MERFISH allows for the simultaneous measurement of a large fraction of the transcriptome in individual cells.
- The technique provides both RNA copy number and precise spatial distribution data within the native cellular context.
Conclusions:
- MERFISH significantly expands the capabilities of smFISH for transcriptomic analysis.
- This method has the potential to revolutionize the study of cellular and tissue functions in health and disease.
