ArticleBioTechniques2018
Vital ex vivo tissue labeling and pathology-guided micropunching to characterize cellular heterogeneity in the tissue microenvironment.
Article in BioTechniques, 2018. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 6 papers.
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Who cites it
6 citing papers in PubMed.
- Differential timing of mitochondrial activation in rat dorsal striatum induced by procedural learning and swimming.Frontiers in molecular neuroscience · 2024Article
- Fresh tissue procurement and preparation for multicompartment and multimodal analysis of the prostate tumor microenvironment.The Prostate · 2022Article
- SEEMLIS: a flexible semi-automated method for enrichment of methylated DNA from low-input samples.Clinical epigenetics · 2022Article
- Harnessing the Utility ofFrontiers in oncology · 2022Review
- Immunofluorescence staining of live lymph node tissue slices.Journal of immunological methods · 2019Article
- 3D organ models-Revolution in pharmacological research?Pharmacological research · 2019Review
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Authors and funding
7 authors.
Funding
Abstract
Cellular heterogeneity within the tissue microenvironment may underlie chemotherapeutic resistance and response, enabling tumor evolution; however, this heterogeneity it is difficult to characterize. Here, we present a new approach-pathology-guided micropunching (PGM)-that enables identification and characterization of heterogeneous foci identified in viable human and animal model tissue slices. This technique consists of live-cell tissue labeling using fluorescent antibodies/small molecules to identify heterogeneous foci (e.g., immune infiltrates or cells with high levels of reactive oxygen species) in viable tissues, coupled with a micropunch step to isolate cells from these heterogeneous foci for downstream molecular or vital functional analysis. Micropunches obtained from epithelial or stromal fibroblast foci in human prostate tissue show 6- to 12-fold enrichment in transcripts specific for EpCam/cytokeratin 8 and vimentin/a-smooth muscle actin/integrin 1-α, respectively. Transcriptional enrichment efficiency agrees with epithelial and stromal laser capture microdissection samples isolated from human prostate. Micropunched foci show a loss of cellular viability in the periphery, but centrally localized cells retained viability before and after dissociation and grew out in culture.
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Registered trials
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