Related Experiment Videos
Immunofluorescence and glutaraldehyde fixation. A new procedure based on the Schiff-quenching method
P Tagliaferro1, C J Tandler, A J Ramos
1Instituto de Biología Celular y Neurociencias Prof. Eduardo De Robertis, Facultad de Medicina, Universidad de Buenos Aires, Argentina.
Journal of Neuroscience Methods
|March 7, 1998
Summary
A new method uses Schiff reagent and sodium borohydride to quench glutaraldehyde autofluorescence in tissues. This enables immunofluorescence staining on glutaraldehyde-fixed samples, expanding research possibilities.
Area of Science:
- Immunology
- Neurobiology
- Biochemistry
Background:
- Immunofluorescence is crucial for antigen localization in tissues, including the central nervous system.
- Formaldehyde is a common fixative, but glutaraldehyde is needed for small molecules (haptens) in neurobiology.
- Glutaraldehyde fixation causes autofluorescence, hindering specific immunofluorescence staining.
Purpose of the Study:
- To develop a method for immunofluorescence on glutaraldehyde-fixed tissues.
- To overcome the limitation of glutaraldehyde-induced autofluorescence.
- To enable the use of glutaraldehyde-raised antisera in immunofluorescence.
Main Methods:
- Tissue sections were treated with Schiff reagent (leucobasic fuchsin).
- The Schiff dye was subsequently reduced using sodium borohydride.
- This process effectively quenched glutaraldehyde-induced tissue autofluorescence.
Main Results:
- The new method successfully reduced glutaraldehyde-induced autofluorescence.
- Specific immunofluorescence staining became observable on glutaraldehyde-fixed tissues.
- The technique allows for the application of previously unusable glutaraldehyde-raised antisera.
Conclusions:
- A novel method allows immunofluorescence on glutaraldehyde-fixed tissues by reducing autofluorescence.
- This technique expands the utility of immunofluorescence in neurobiology and other fields.
- It facilitates the use of a wider range of antibodies for basic and clinical research.