Laser Capture Microscopy Coupled with Smart-seq2 for Precise Spatial Transcriptomic Profiling

Overview

Journal Nat Commun

Specialty Biology

Date 2016 Jul 9

PMID 27387371

Citations 151

Authors

Susanne Nichterwitz

Geng Chen

Julio Aguila Benitez

Marlene Yilmaz

Helena Storvall

Ming Cao

Rickard Sandberg

Qiaolin Deng

Eva Hedlund

Affiliations

Soon will be listed here.

Abstract

Laser capture microscopy (LCM) coupled with global transcriptome profiling could enable precise analyses of cell populations without the need for tissue dissociation, but has so far required relatively large numbers of cells. Here we report a robust and highly efficient strategy for LCM coupled with full-length mRNA-sequencing (LCM-seq) developed for single-cell transcriptomics. Fixed cells are subjected to direct lysis without RNA extraction, which both simplifies the experimental procedures as well as lowers technical noise. We apply LCM-seq on neurons isolated from mouse tissues, human post-mortem tissues, and illustrate its utility down to single captured cells. Importantly, we demonstrate that LCM-seq can provide biological insight on highly similar neuronal populations, including motor neurons isolated from different levels of the mouse spinal cord, as well as human midbrain dopamine neurons of the substantia nigra compacta and the ventral tegmental area.

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