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Laser Microdissection Applied to Gene Expression Profiling of Subset of Cells from the Drosophila Wing Disc

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Methods for Selective Gene Expression Profiling in Single Tertiary Lymphoid Structure Using Laser Capture

Claudia Gutierrez-Chavez^1,2,3,4, Samantha Knockaert^1,2,3,5, Marie-Caroline Dieu-Nosjean^6,7,8

¹Institut National de la Santé et de la Recherche Médicale (INSERM), UMRS 1138, Cordeliers Research Center, Laboratory "Cancer, Immune Control and Escape", Paris, France.

Methods in Molecular Biology (Clifton, N.J.)

|November 11, 2024

Summary

Tertiary lymphoid structures (TLS) form during inflammation and drive local immune responses. This study details a laser capture microdissection (LCM) method for isolating TLS in situ for molecular analysis.

Keywords:

Frozen tissue Gene expression profiling Laser capture microdissection Lung cancer Lymphocyte RNA analysis Tertiary lymphoid structure

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Last Updated: Jun 7, 2025

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Area of Science:

Immunology
Pathology
Molecular Biology

Background:

Tertiary lymphoid structures (TLS) are dynamic, de novo lymphoid formations in tissues during inflammation.
TLS mimic secondary lymphoid organs (SLOs) and are implicated in cancer, infections, autoimmunity, and transplant rejection.
TLS significantly influence local adaptive immune responses.

Purpose of the Study:

To describe a detailed laser capture microdissection (LCM) method for the in situ isolation of TLS.
To enable comprehensive downstream molecular characterization of TLS and their microenvironment.

Main Methods:

Utilizing laser capture microdissection (LCM) for precise isolation of TLS from tissue sections.
Application of LCM to both frozen and paraffin-embedded tissue samples.
Focus on in situ isolation to preserve tissue architecture and cellular context.

Main Results:

Demonstrated the feasibility of LCM for isolating intact TLS.
Enabled the selection of specific cellular components within TLS.
Facilitated molecular profiling of TLS and their associated microenvironment.

Conclusions:

LCM is a powerful technique for studying TLS in their native tissue context.
This method allows for detailed molecular analysis of TLS, advancing understanding of their role in disease.
The described protocol aids in deciphering the complex interactions within the local immune microenvironment.