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Laser capture microscopy coupled with Smart-seq2 for precise spatial transcriptomic profiling.

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.

SUBMITTER: Nichterwitz S 

PROVIDER: S-EPMC4941116 | biostudies-literature | 2016 Jul

REPOSITORIES: biostudies-literature

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Laser capture microscopy coupled with Smart-seq2 for precise spatial transcriptomic profiling.

Nichterwitz Susanne S   Chen Geng G   Aguila Benitez Julio J   Yilmaz Marlene M   Storvall Helena H   Cao Ming M   Sandberg Rickard R   Deng Qiaolin Q   Hedlund Eva E  

Nature communications 20160708

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  ...[more]

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