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Pixelated spatial gene expression analysis from tissue.
- Source :
-
Nature communications [Nat Commun] 2018 Jan 15; Vol. 9 (1), pp. 202. Date of Electronic Publication: 2018 Jan 15. - Publication Year :
- 2018
-
Abstract
- Here, we present a technique that performs on-chip picoliter real-time reverse transcriptase loop mediated isothermal amplification (RT-LAMP) reactions on a histological tissue section without any analyte purification while preserving the native spatial location of the nucleic acid molecules. We demonstrate this method by amplifying TOP2A messenger RNA (mRNA) in a prostate cancer xenograft with 100 µm spatial resolution and by visualizing the variation in threshold time of amplification across the tissue. The on-chip reaction was validated by mRNA fluorescence in situ hybridization (mFISH) from cells in the tissue section. The entire process, from tissue loading on microchip to results from RT-LAMP can be carried out in less than 2 h. We anticipate that this technique, with its ease of use, fast turnaround, and quantitative molecular outputs, would become an invaluable tissue analysis tool for researchers and clinicians in the biomedical arena.
- Subjects :
- Animals
Cell Line, Tumor
DNA Topoisomerases, Type II genetics
Heterografts
Humans
In Situ Hybridization, Fluorescence
Male
Mice
Mice, Nude
Microchip Analytical Procedures
Poly-ADP-Ribose Binding Proteins genetics
RNA, Messenger genetics
Real-Time Polymerase Chain Reaction
Reproducibility of Results
Reverse Transcriptase Polymerase Chain Reaction
Spectroscopy, Fourier Transform Infrared
Gene Expression Profiling
Prostatic Neoplasms genetics
Subjects
Details
- Language :
- English
- ISSN :
- 2041-1723
- Volume :
- 9
- Issue :
- 1
- Database :
- MEDLINE
- Journal :
- Nature communications
- Publication Type :
- Academic Journal
- Accession number :
- 29335461
- Full Text :
- https://doi.org/10.1038/s41467-017-02623-9