Document Type

Article

Publication Date

12-16-2022

Publication Title

STAR Protoc

Keywords

JGM, Animals, Humans, Mice, Fluorescent Antibody Technique, Gene Expression Profiling, Laser Capture Microdissection, Melanoma, Transcriptome

JAX Source

STAR Protoc. 2022;3(4):101698.

Volume

3

Issue

4

First Page

101698

ISSN

2666-1667

PMID

36149794

DOI

https://doi.org/10.1016/j.xpro.2022.101698

Grant

We thank our patients and tissue donors; The Microscopy, Single Cell Biology, Genome Technology, and CTRS Scientific Services of JAX partially supported through the NCI P30 CA034196; supported by The Jackson Laboratory and NCI (R01 CA195712, R01 CA204115).

Abstract

We describe a pipeline for optimized and streamlined multiplexed immunofluorescence-guided laser capture microdissection allowing the harvest of individual cells based on their phenotype and tissue localization for transcriptomic analysis with next-generation RNA sequencing. Here, we analyze transcriptomes of CD3+ T cells, CD14+ monocytes/macrophages, and melanoma cells in non-dissociated metastatic melanoma tissue. While this protocol is described for melanoma tissues, we successfully applied it to human tonsil, skin, and breast cancer tissues as well as mouse lung tissues. For complete details on the use and execution of this protocol, please refer to Martinek et al. (2022).

Comments

This is an open access article under the CC BY-NC-ND license.

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