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Immunofluorescence Microscopy

Immunofluorescence Microscopy

A fluorescence microscope uses fluorescent chromophores called fluorochromes, which can absorb energy from a light source and then emit this energy as visible light. Fluorochromes include naturally fluorescent substances (such as chlorophylls) and fluorescent stains that are added to the specimen to create contrast. Dyes such as Texas red and FITC are examples of fluorochromes. Other examples include the nucleic acid dyes 4’,6’-diamidino-2-phenylindole (DAPI), and acridine orange.

Immunocytochemistry and Immunohistochemistry

Immunocytochemistry and Immunohistochemistry

Immunocytochemistry (ICC) and immunohistochemistry (IHC) are techniques that use antibodies to check for specific proteins or antigens in a sample. The technique was first published by Albert Coons in 1941 to detect the presence of pneumococcal antigen in tissue sections from mice infected with Pneumococcus. Immunocytochemistry helps localization of proteins or antigens in individual cells like blood cells, stem cells, etc., while immunohistochemistry does the same for tissue samples.
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Related Experiment Video

Updated: Dec 29, 2025

Immunofluorescence to Monitor the Cellular Uptake of Human Lactoferrin and its Associated Antiviral Activity Against the Hepatitis C Virus

Immunofluorescence to Monitor the Cellular Uptake of Human Lactoferrin and its Associated Antiviral Activity Against the Hepatitis C Virus

Published on: October 1, 2015

Practical Direct Immunofluorescence.

Randie H Kim¹, Nooshin K Brinster²

¹Assistant Professor, The Ronald O. Perelman Department of Dermatology, New York University School of Medicine, New York, NY; and.

The American Journal of Dermatopathology

|January 29, 2020

Summary

This summary is machine-generated.

Direct immunofluorescence (DIF) is a valuable diagnostic tool for autoimmune blistering disorders and vasculitis. This review offers practical guidance on specimen handling and interpretation to improve DIF utilization in clinical practice.

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Area of Science:

Immunopathology
Dermatology
Clinical Diagnostics

Background:

Direct immunofluorescence (DIF) is an essential diagnostic technique.
Its full potential may be limited by perceived challenges in interpretation, specimen processing, and handling.
Standardized protocols are needed to optimize its use.

Purpose of the Study:

To provide a practical guide for the appropriate use of DIF in various clinical diseases.
To address challenges associated with DIF interpretation, limitations, and specimen processing.
To highlight the evolving role of DIF in specific conditions like vasculitis and connective tissue diseases.

Main Methods:

Review of current literature and clinical guidelines on DIF.
Discussion of practical aspects including biopsy technique, specimen handling, and storage.
Analysis of DIF applications in autoimmune blistering disorders, vasculitis, and connective tissue diseases.

Main Results:

DIF is crucial for diagnosing autoimmune blistering disorders.
Its role in IgA vasculitis is evolving, offering diagnostic value.
DIF can be beneficial in connective tissue diseases with negative serologies or nondiagnostic histology, particularly for scarring alopecia.

Conclusions:

Optimizing DIF specimen handling and interpretation can enhance its clinical utility.
DIF remains a key tool for specific dermatological and rheumatological conditions.
Further research and standardized training can address current limitations and improve diagnostic accuracy.