Laser capture microdissection--a demonstration of the isolation of individual dopamine neurons and the entire ventral tegmental area

Journal of Visualized Experiments : JoVE
Evangel KummariJeffrey B Eells

Abstract

Laser capture microdissection (LCM) is used to isolate a concentrated population of individual cells or precise anatomical regions of tissue from tissue sections on a microscope slide. When combined with immunohistochemistry, LCM can be used to isolate individual cells types based on a specific protein marker. Here, the LCM technique is described for collecting a specific population of dopamine neurons directly labeled with tyrosine hydroxylase immunohistochemistry and for isolation of the dopamine neuron containing region of the ventral tegmental area using indirect tyrosine hydroxylase immunohistochemistry on a section adjacent to those used for LCM. An infrared (IR) capture laser is used to both dissect individual neurons as well as the ventral tegmental area off glass slides and onto an LCM cap for analysis. Complete dehydration of the tissue with 100% ethanol and xylene is critical. The combination of the IR capture laser and the ultraviolet (UV) cutting laser is used to isolate individual dopamine neurons or the ventral tegmental area when using PEN membrane slides. A PEN membrane slide has significant advantages over a glass slide as it offers better consistency in capturing and collecting cells, is faster collecting lar...Continue Reading

Citations

Nov 9, 2016·Frontiers in Cell and Developmental Biology·Ping HuGlenn Deng
Jun 27, 2020·Cells·Pallavi GuptaTuhin Subhra Santra
Jun 1, 2017·Histochemistry and Cell Biology·P Garrido-GilJose L Labandeira-Garcia
Mar 23, 2021·Frontiers in Molecular Neuroscience·Jordan SichermanShreejoy J Tripathy

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Methods Mentioned

BETA
Laser Capture Microdissection
dissections
dissection

Software Mentioned

Microscope
LCM

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