Name: Laser Capture Microdissection of Mammalian Tissue (Video) | JoVE Journal
Uploaded: 2007-10-01T00:00:00.000+00:00
Duration: 16 min 34 s
Description: Learn laser capture microdissection using the Leica A-S-L-M-D microscope to isolate specific mammalian tissue samples with high precision. This technique enables accurate RNA isolation and enhances biological research quality.

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laser capture microdissection of mammalian tissue

Laser Capture Microdissection of Mammalian Tissue

Laser capture microscopy, also known as laser microdissection (LMD), enables the user to isolate small numbers of cells or tissues from frozen or formalin-fixed, paraffin-embedded tissue sections. LMD techniques rely on a thermo labile membrane placed either on top of, or underneath, the tissue section. In one method, focused laser energy is used to melt the membrane onto the underlying cells, which can then be lifted out of the tissue section. In the other, the laser energy vaporizes the foil along a path "drawn" on the tissue, allowing the selected cells to fall into a collection device. Each technique allows the selection of cells with a minimum resolution of several microns. DNA, RNA, protein, and lipid samples may be isolated and analyzed from micro-dissected samples. In this video, we demonstrate the use of the Leica AS-LMD laser microdissection instrument in seven segments, including an introduction to the principles of LMD, initializing the instrument for use, general considerations for sample preparation, mounting the specimen and setting up capture tubes, aligning the microscope, adjusting the capture controls, and capturing tissue specimens. Laser-capture micro-dissection enables the investigator to isolate samples of pure cell populations as small as a few cell-equivalents. This allows the analysis of cells of interest that are free of neighboring contaminants, which may confound experimental results.

Laser capture microscopy, also known as laser microdissection (LMD), enables the user to isolate small numbers of cells or tissues from frozen or ...

Hi, my name is Rob Edwards. I'm the director of the University of California Irvine Laser Capture microscopy facility. I'm going to be describing for you the use of the laser capture microscope that we have here.Our instrument is a Leica A-S-L-M-D, which we purchased approximately three years ago. There are two different types of laser capture systems. One can be likened to a spot welder where one has a piece of tissue on a glass slide on top of a plastic membrane, which overlies the tissue.A laser source is then used to melt the membrane, in effect spot welding the tissue onto the membrane and then that membrane is removed, taking the captured tissue with it.Okay. To start the LCM system, we power on a number of instruments in sequence here. We'll start with the A-S-L-M-D control box, which is powered on here.We then turn on the computer simple enough. If you have a specimen that requires a fluorescence optics for identifying the cells to be captured, you would turn on the mercury lamp for the fluorescence illumination. We're not going to be using that for this demonstration.If you want to capture high quality images of the before and after specimens, one can turn on the spot came

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Research

JoVE Journal

Biology

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Laser Microdissection: A Technique to Isolate Invading Colorectal Cancer cells from a 3D Organotypic Culture Model

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A Laser Capture Microdissection Protocol That Yields High Quality RNA from Fresh-frozen Mouse Bones

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Laser Capture Microdissection of Mouse Embryonic Cartilage and Bone for Gene Expression Analysis

Laser microdissection (LMD) is a sensitive and highly reproducible technique that can be used to uncover pathways that mediate glioma heterogeneity and invasion. Here, we describe an optimized protocol to isolate discrete areas from glioma tissue using laser LMD followed by transcriptomic analysis.

Laser Capture Microdissection of Glioma Subregions for Spatial and Molecular Characterization of Intratumoral Heterogeneity, Oncostreams, and Invasion

A protocol is described that uses laser microdissection to isolate individual nematode tissues for RNA-sequencing. The protocol does not require species-specific genetic toolkits, allowing gene expression profiles to be compared between different species at the level of single-tissue samples.

Laser Microdissection for Species-Agnostic Single-Tissue Applications

This protocol describes a high-throughput workflow for artificial intelligence-driven segmentation of pathology-confirmed regions of interest from stained, thin tissue section images for enrichment of histology-resolved cell populations using laser microdissection. This strategy includes a novel algorithm enabling the transfer of demarcations denoting cell populations of interest directly to laser microscopes.

Industrialized, Artificial Intelligence-guided Laser Microdissection for Microscaled Proteomic Analysis of the Tumor Microenvironment

This protocol describes a method to isolate RNA from defined histological regions of paraformaldehyde-fixed, Optimal Cutting Temperature compound-embedded mouse liver tissue using laser capture microdissection, enabling targeted downstream gene expression analysis.

Laser Capture Microdissection of Paraformaldehyde-Fixed Mouse Liver Tissue for RNA Analysis

Learn laser capture microdissection using the Leica A-S-L-M-D microscope to isolate specific mammalian tissue samples with high precision. This technique enables accurate RNA isolation and enhances biological research quality.

Laser Capture Microdissection of Mammalian Tissue

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