LCM-Seq: A Method for Spatial Transcriptomic Profiling Using Laser Capture Microdissection Coupled with PolyA-Based RNA Sequencing

Methods Mol Biol. 2018;1649:95-110. doi: 10.1007/978-1-4939-7213-5_6.

Abstract

LCM-seq couples laser capture microdissection of cells from frozen tissues with polyA-based RNA sequencing and is applicable to single neurons. The method utilizes off-the-shelf reagents and direct lysis of the cells without RNA purification, making it a simple and relatively cheap method with high reproducibility and sensitivity compared to previous methods. The advantage with LCM-seq is also that tissue sections are kept intact and thus the positional information of each cell is preserved.

Keywords: Cresyl violet; Dopamine neuron; HistoGene; Laser capture microscopy; Motor neuron; RNA sequencing; Rapid antibody staining; Smart-seq2.

MeSH terms

  • Animals
  • Antibodies / metabolism
  • Cell Separation
  • Cells, Cultured
  • DNA, Complementary / genetics
  • Gene Expression Profiling / methods*
  • Gene Library
  • Humans
  • Laser Capture Microdissection / methods*
  • Mice
  • Poly A / metabolism*
  • Sequence Analysis, RNA / methods*
  • Staining and Labeling
  • Tyrosine 3-Monooxygenase / metabolism

Substances

  • Antibodies
  • DNA, Complementary
  • Poly A
  • Tyrosine 3-Monooxygenase