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

IF: 17.694

Cited by: 166

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.

Keywords

LCM-seq

Spatial Transcriptomics

MeSH terms

Animals

Dopaminergic Neurons

Female

Gene Expression

Gene Expression Profiling

Humans

Laser Capture Microdissection

Male

Mesencephalon

Mice

Microscopy

Models, Animal

Motor Neurons

Mouse Embryonic Stem Cells

Pars Compacta

Poly A

RNA, Messenger

Sequence Analysis, RNA

Spinal Cord

Ventral Tegmental Area

Authors

Nichterwitz, Susanne

Chen, Geng

Aguila Benitez, Julio

Yilmaz, Marlene

Storvall, Helena

Cao, Ming

Sandberg, Rickard

Deng, Qiaolin

Hedlund, Eva

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