ArticleScientific reports2020
An Optimised Protocol Harnessing Laser Capture Microdissection for Transcriptomic Analysis on Matched Primary and Metastatic Colorectal Tumours.
Article in Scientific reports, 2020. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 12 papers.
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Who cites it
12 citing papers in PubMed, 20 citations in OpenAlex.
- Feed restriction affects follicular development by regulating cell cycle progression in porcine mural granulosa cells.Journal of animal science and biotechnology · 2026Article
- Immuno-laser capture microdissection of perfusion-fixed mouse brain tissue coupled to RNA-seq.Journal of neuroscience methods · 2025Article
- Transcriptomic Signatures in IgA Nephropathy: From Renal Tissue to Precision Risk Stratification.International journal of molecular sciences · 2025Review
- Integration of hyperspectral imaging and transcriptomics from individual cells with SpectralSeq.Genome research · 2025Article
- Extraction, Purification, and Next-Generation Sequencing (NGS) Analysis of DNA and RNA from Formalin-Fixed and Paraffin-Embedded (FFPE) Tissue.Methods in molecular biology (Clifton, N.J.) · 2025Article
- An optimized workflow for transcriptomic analysis from archival paraformaldehyde-fixed retinal tissues collected by laser capture microdissection.Experimental eye research · 2024Article
- Comparative transcriptomics reveals highly conserved regional programs between porcine and human colonic enteric nervous system.Communications biology · 2023Article
- Single-cell RNA-sequencing of retrieved human oocytes and eggs in clinical practice and for human ovarian cell atlasing.Molecular reproduction and development · 2022Review
- Transcriptomics and solid tumors: The next frontier in precision cancer medicine.Seminars in cancer biology · 2022Review
- Opioid Receptor Expression in Colorectal Cancer: A Nested Matched Case-Control Study.Frontiers in oncology · 2022Article
- Shape decomposition algorithms for laser capture microdissection.Algorithms for molecular biology : AMB · 2021Article
- Article
Corrections and comments
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Authors and funding
20 authors at 4 institutions in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Generation of large amounts of genomic data is now feasible and cost-effective with improvements in next generation sequencing (NGS) technology. Ribonucleic acid sequencing (RNA-Seq) is becoming the preferred method for comprehensively characterising global transcriptome activity. Unique to cytoreductive surgery (CRS), multiple spatially discrete tumour specimens could be systematically harvested for genomic analysis. To facilitate such downstream analyses, laser capture microdissection (LCM) could be utilized to obtain pure cell populations. The aim of this protocol study was to develop a methodology to obtain high-quality expression data from matched primary tumours and metastases by utilizing LCM to isolate pure cellular populations. We demonstrate an optimized LCM protocol which reproducibly delivered intact RNA used for RNA sequencing and quantitative polymerase chain reaction (qPCR). After pathologic annotation of normal epithelial, tumour and stromal components, LCM coupled with cDNA library generation provided for successful RNA sequencing. To illustrate our framework's potential to identify targets that would otherwise be missed with conventional bulk tumour sequencing, we performed qPCR and immunohistochemical technical validation to show that the genes identified were truly expressed only in certain sub-components. This study suggests that the combination of matched tissue specimens with tissue microdissection and NGS provides a viable platform to unmask hidden biomarkers and provides insight into tumour biology at a higher resolution.
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