Fluorescent Stain for Cell Necrosis

April 10, 2019

BlueskyReddit
Physiology
Jali Medical Inc.
Research Scientist

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Company:

Histo-Chem, Inc.

Product Name:

Fluoro-Jade B

Catalog Number:

1FJB

 

 

As a complementary technique to Nissl and H&E stains, the Fluoro-Jade B stain, which is an anionic fluorescein derivative, is a method of detecting active neuronal degeneration. The stain works well to visualize and help quantify cell necrosis after brain injury in animal models.

Experimental Design and Results Summary

Applications

Immunofluorescence

Sample

Mouse neocortex brain tissue

Primary Incubation

Immerse mounted slides in 100% Ethanol for 5 minutes and then in 70% ethanol for 2 minutes

Blocking Agent

Gently shake in 0.06% Potassium permanganate for 15 minutes

Secondary Incubation

Stain slides in 0.001% Fluoro-Jade B in 0.1% acetic acid solution (gently shaking) for 30min in the dark

Tertiary Incubation

N/A

Detection

Fluorescence visualized with a confocal microscope

Results Summary

Fluoro-Jade B staining of neocortical slices involved a simple protocol where tissue slices mounted on slides are exposed to a series of solutions for clearing, permeabilization and background blocking. Very little Fluoro-Jade B is required to stain 20-50 slides with mounted tissue. This allows visualization of cells that are in the the process of dying and offers a means for quantifying degeneration. The only issue is tissue loss due to tissue falling off slides as the slides are transferred into different solutions.

DOI or PMID #

N/A

Additional Notes

Recommend using Superfrost Plus Microscope Slides

Related Categories

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Summary

The Good

Simple method for visualizing neuronal degeneration

The Bad

Neuronal processes were not easily detected

The Bottom Line

The Fluoro-Jade B stain can be used as a complimentary method or alternative method to visual injury induced cell necrosis. The amount of product needed is very little for visualization and the methodology is very straight forward. The only problematic requirement is that the procedure requires sample tissue mounted on slides as a first step.

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