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Laser-Capture Microdissection RNA-Sequencing for Spatial and Temporal Tissue-Specific Gene Expression Analysis in Plants
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Laser-Capture Microdissection RNA-Sequencing for Spatial and Temporal Tissue-Specific Gene Expression Analysis in Plants

Laser-Capture Microdissection RNA-Sequencing for Spatial and Temporal Tissue-Specific Gene Expression Analysis in Plants

DOI:
10.3791/61517-v

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08:33 min

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August 05, 2020

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, , , , ,
1Department of Animal, Plant and Soil Science, AgriBio Building,La Trobe University, 2Australian Research Council Centre of Excellence in Plant Energy Biology,La Trobe University, 3Australian Research Council Research Hub for Medicinal Agriculture, AgriBio Building,La Trobe University

Chapters

  • 00:04Introduction
  • 01:11Tissue Fixation
  • 01:47Tissue Processing
  • 02:38Paraffin Embedding
  • 03:21Polyethylene Naphthalate (PEN) Membrane Slide Preparation
  • 03:49Sectioning
  • 04:53Laser-Capture Microdissection (LCM)
  • 06:14Results: Representative LCM Sample Acquisition and RNA Quantification
  • 07:52Conclusion

Summary

Automatic Translation

Automatic Translation

Presented here is a protocol for laser-capture microdissection (LCM) of plant tissues. LCM is a microscopic technique for isolating areas of tissue in a contamination-free manner. The procedure includes tissue fixation, paraffin embedding, sectioning, LCM and RNA extraction. RNA is used in the downstream tissue-specific, temporally resolved analysis of transcriptomes.

Tags

Laser-capture MicrodissectionRNA-sequencingSpatial TranscriptomeTemporal TranscriptomePlant CellsParaffin EmbeddingBarley SeedVacuum InfiltrationDehydrationXyleneParaffin Infiltration

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