Jove
Visualize
Contact Us
JoVE
x logofacebook logolinkedin logoyoutube logo
ABOUT JoVE
OverviewLeadershipBlogJoVE Help Center
AUTHORS
Publishing ProcessEditorial BoardScope & PoliciesPeer ReviewFAQSubmit
LIBRARIANS
TestimonialsSubscriptionsAccessResourcesLibrary Advisory BoardFAQ
RESEARCH
JoVE JournalMethods CollectionsJoVE Encyclopedia of ExperimentsArchive
EDUCATION
JoVE CoreJoVE BusinessJoVE Science EducationJoVE Lab ManualFaculty Resource CenterFaculty Site
Terms & Conditions of Use
Privacy Policy
Policies

Related Experiment Videos

Improved indirect fluorescence immunocytochemical method using counterstains.

R F Newkirk1, J Mack

  • 1Department of Biological Sciences, Tennessee State University, Nashville 37209-1561.

Biotechniques
|October 1, 1992
PubMed
Summary
This summary is machine-generated.

Related Concept Videos

You might also read

Related Articles

Articles linked to this work by shared authors, journal, and citation graph.

Sort by
Same author

From chaos to tranquillity: a modern approach to the identification, nomenclature and phylogeny of <i>Aspergillus</i>, <i>Penicillium</i> and other <i>Eurotiales</i>, including an updated accepted species list.

Studies in mycology·2026
Same author

Genetics by genetic algorithm: Defining an ideal platelet donor population to support patients with HLA-mediated alloimmune refractoriness.

Transfusion·2025
Same author

Fungal Planet description sheets: 1478-1549.

Persoonia·2024
Same author

Taxonomy and phylogeny of the basidiomycetous hyphomycete genus <i>Hormomyces</i>.

Fungal systematics and evolution·2021
Same author

Faecal calcium excretion does not decrease during long-term feeding of a low-calcium diet in adult dogs.

Journal of animal physiology and animal nutrition·2017
Same author

Mast cells in the human dura: effects of age and dural bleeding--authors' reply.

Child's nervous system : ChNS : official journal of the International Society for Pediatric Neurosurgery·2013
Same journal

Investigating the interactomic landscape of survival motor neuron (SMN) and the SMNΔ7 truncated protein.

BioTechniques·2026
Same journal

Antigen retrieval-immunofluorescence on free floating sections to visualize the liver lobule and its cellular makeup.

BioTechniques·2026
Same journal

Special approach of droplet digital polymerase chain reaction (ddPCR) for transgene stability of a Chinese hamster ovary (CHO) cell line.

BioTechniques·2026
Same journal

Strand-specific quantification of L1 ORF0 and related transcripts by multiplex reverse transcription with tagged primers.

BioTechniques·2026
Same journal

Why and when should we choose digital PCR?

BioTechniques·2026
Same journal

Quantitative and unbiased lung alveolar septum assessment in an LPS experimental mouse model using 2D-spatial correlation image analysis from hematoxylin and eosin slides.

BioTechniques·2026
See all related articles

This study introduces Evans blue as a counterstain to improve indirect immunocytochemistry. It enhances cell localization and significantly prolongs fluorescein isothiocyanate (FITC) fluorescence, overcoming common technique limitations.

Area of Science:

  • Neurobiology
  • Immunocytochemistry
  • Fluorescence Microscopy

Background:

  • Indirect immunofluorescence techniques are valuable in neurobiology but suffer from poor localization and fading fluorescence.
  • Commonly used fluorophores include fluorescein isothiocyanate (FITC) and tetrahodamine isothiocyanate (TRITC).

Purpose of the Study:

  • To modify indirect immunocytochemical techniques to enhance cell localization and prolong fluorescence.
  • To evaluate the efficacy of Evans blue as a counterstain in conjunction with FITC, TRITC, and 7-amino-4-methyl-coumarin-3-acetic acid (AMCA).

Main Methods:

  • An indirect immunocytochemical technique was modified using FITC, TRITC, and AMCA.
  • Evans blue was employed as a counterstain to assess its impact on localization and fluorescence.

Related Experiment Videos

  • Compatibility of Evans blue with different fluorophores was tested.

    • Main Results:

    • Evans blue counterstaining improved cell localization for FITC and AMCA, providing a clear morphological background.
    • Evans blue significantly extended the fluorescence lifetime of FITC from days to weeks.
    • Evans blue was found to be incompatible with TRITC.

    Conclusions:

    • Evans blue is an effective counterstain for improving indirect fluorescence immunocytochemistry with FITC and AMCA.
    • The use of Evans blue enhances both cell localization and fluorescence stability, addressing key limitations of the technique.