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Drosophila Larval IHC

Overzicht

Immunohistochemistry (IHC) is a technique used to visualize the presence and location of proteins within tissues. Drosophila larvae are particularly amenable to IHC because of the ease with which they can be processed for staining. Additionally, the larvae are transparent, meaning that some tissues can be visualized without the need for dissection.

In IHC, proteins are ultimately detected with antibodies that specifically bind to “epitopes” within the protein of interest. In order to preserve these epitopes, tissues must be fixed prior to staining. Furthermore, in order for antibodies to penetrate membranes, cells must be permeabilized with detergents. This video article offers a detailed view of the reagents, tools, and procedures necessary for the staining of dissected larval tissues, including fixation, blocking, and staining steps. Also featured is a demonstration of tissue mounting techniques for fluorescence microscopy. Finally, examples of the broad range of applications for these techniques (and some variations upon them) are provided.

Procedure

Drosophila melanogaster is a widely studied model organism due to its versatility and facility of use.

Immunohistochemistry performed on Drosophila larval preparations is an invaluable method that can provide information on the presence, location, and the co-localization of proteins.

This video will cover the essential methods for the dissection, fixation, blocking, and mounting of Drosophila larval tissue and examples of their applications.

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Tags

Drosophila MelanogasterModel OrganismImmunohistochemistryDrosophila Larval IHCDrosophilaLarval PreparationsProteinsDissectionFixationBlockingMountingAntibodiesVisualizationTissuePhosphate Buffered Saline PBSOrgansSaline SolutionPHFormaldehyde based SolutionPreservationEnzymatic BreakdownWashing StepsImmunostaining ProcessTriton X100

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0:00

Overview

0:41

Principles of Larval Immunohistochemistry

3:29

Immunohistochemistry Procedure

5:34

Applications

8:03

Summary

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