Search research articles

ABOUT JoVE

Overview Leadership Blog JoVE Help Center

AUTHORS

Publishing Process Editorial Board Scope & Policies Peer Review FAQ Submit

LIBRARIANS

Testimonials Subscriptions Access Resources Library Advisory Board FAQ

RESEARCH

JoVE Journal Methods Collections JoVE Encyclopedia of Experiments Archive

EDUCATION

JoVE Core JoVE Business JoVE Science Education JoVE Lab Manual Faculty Resource Center Faculty Site

Terms & Conditions of Use

Related Concept Videos

Fixation and Sectioning

Fixation and Sectioning

Two basic types of preparation are used to visualize specimens with a light microscope: wet mounts and fixed specimens.
The simplest type of preparation is the wet mount, in which the specimen is placed in a drop of liquid on the slide. A liquid specimen can be directly deposited on the slide using a dropper. Solid specimens, such as skin scraping, can be placed on the slide before adding a drop of liquid to prepare the wet mount. Sometimes the liquid is simply water, but stains are often added...

You might also read

Related Articles

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

Sort by

Same author

The Epidermal Growth Factor Receptor (EGFR) as a means of neuro-immune cross-conversation.

Journal of neuroinflammation·2026

Same author

Progressive HNF1A-MODY pathophysiology revealed by a translational mouse model.

JCI insight·2026

Same author

Gene delivery of immunomodulatory cytokines to the lung preserves respiratory function during inflammatory challenge.

Science immunology·2026

Same author

Adult patients with autoinflammation of unknown origin partially phenocopy the immune presentation of Still's disease.

Nature communications·2026

Same author

TRACKing the tissue-specific origins of diverse effector T cell functions.

Nature immunology·2026

Same author

IL-2 silencing enables Tfh cell expansion during vaccination but is redundant for antibody production.

Immunology and cell biology·2026

Same journal

A Novel Laboratorial Approach to Evaluate Bacterial Microleakage of Endodontic Sealers.

Current protocols·2026

Same journal

TRIAGE Toolkit: Streamlined Discovery of Regulatory Genes and Elements.

Current protocols·2026

Same journal

High-throughput Profiling of Pseudouridines in Microbiome-derived Bacterial RNA.

Current protocols·2026

Same journal

Recombinant Protein Expression in Rhodococcus species.

Current protocols·2026

Same journal

Streamlined In Vitro Transcription for Generating Self-Amplifying RNA With Modified Nucleotides.

Current protocols·2026

Same journal

CODEC Library Preparation From Genomic DNA.

Current protocols·2026

See all related articles

Search research articles

Related Experiment Video

Updated: May 2, 2026

Targeted Plasma Membrane Delivery of a Hydrophobic Cargo Encapsulated in a Liquid Crystal Nanoparticle Carrier

Targeted Plasma Membrane Delivery of a Hydrophobic Cargo Encapsulated in a Liquid Crystal Nanoparticle Carrier

Published on: February 8, 2017

Introducing the Dish Soap Protocol: A Unified Approach for Multi-Modal Intracellular Staining.

Oliver T Burton¹, James Dooley^1,2, Adrian Liston^1,2

¹Department of Pathology, University of Cambridge, Cambridge, UK.

Current Protocols

|September 19, 2025

Summary

This summary is machine-generated.

New fixation buffers enable simultaneous intracellular and nuclear staining in flow cytometry, overcoming limitations of current methods for improved cell analysis. This advance supports multicolor flow cytometry applications.

Keywords:

GFP flow cytometry immune phenotyping intracellular staining spectral flow cytometry

More Related Videos

An Automated Differential Nuclear Staining Assay for Accurate Determination of Mitocan Cytotoxicity

An Automated Differential Nuclear Staining Assay for Accurate Determination of Mitocan Cytotoxicity

Published on: May 12, 2020

Automated Imaging and Analysis for the Quantification of Fluorescently Labeled Macropinosomes

Automated Imaging and Analysis for the Quantification of Fluorescently Labeled Macropinosomes

Published on: August 24, 2021

Related Experiment Videos

Last Updated: May 2, 2026

Targeted Plasma Membrane Delivery of a Hydrophobic Cargo Encapsulated in a Liquid Crystal Nanoparticle Carrier

Targeted Plasma Membrane Delivery of a Hydrophobic Cargo Encapsulated in a Liquid Crystal Nanoparticle Carrier

Published on: February 8, 2017

An Automated Differential Nuclear Staining Assay for Accurate Determination of Mitocan Cytotoxicity

An Automated Differential Nuclear Staining Assay for Accurate Determination of Mitocan Cytotoxicity

Published on: May 12, 2020

Automated Imaging and Analysis for the Quantification of Fluorescently Labeled Macropinosomes

Automated Imaging and Analysis for the Quantification of Fluorescently Labeled Macropinosomes

Published on: August 24, 2021

Area of Science:

Cellular Biology
Immunology
Biochemistry

Background:

Multidimensional flow cytometry allows for extensive cellular analysis.
Current fixation protocols present challenges in simultaneously preserving intracellular targets and nuclear antigens.
Balancing membrane permeabilization and structural integrity is crucial for intracellular staining.

Purpose of the Study:

To evaluate the impact of various fixation and permeabilization reagents on key cellular features.
To develop a cost-effective buffer solution for enhanced multidimensional flow cytometry.
To enable simultaneous detection of intracellular proteins and nuclear targets without compromising cell integrity or fluorescence.

Main Methods:

Assessment of fix-perm reagents on nuclear staining, fluorescent protein retention (e.g., GFP), cytokine detection, epitope preservation, and cell viability.
Development and testing of a novel buffer formulation, "Burton's Best Buffer".
Comparison of the novel buffer with commercial fixation buffers.

Main Results:

Common fixation protocols demonstrate trade-offs between nuclear accessibility and intracellular target preservation.
The novel "Burton's Best Buffer" effectively preserves fluorescent proteins and allows for nuclear staining.
The developed buffer maintains cell integrity, scatter profiles, and fluorophore stability while enabling detection of transcription factors and cytokines.
"Burton's Best Buffer" is significantly more cost-effective than commercial alternatives.

Conclusions:

A novel, cost-effective fixation buffer overcomes limitations in multidimensional flow cytometry.
Simultaneous detection of intracellular and nuclear targets is achievable, enhancing cellular analysis.
This protocol supports advanced flow cytometry applications, including cytokine and transcription factor analysis.