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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.
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Related Experiment Video

Updated: Jul 20, 2026

Immunohistological Labeling of Microtubules in Sensory Neuron Dendrites, Tracheae, and Muscles in the Drosophila Larva Body Wall

Immunohistological Labeling of Microtubules in Sensory Neuron Dendrites, Tracheae, and Muscles in the Drosophila Larva Body Wall

Published on: November 10, 2011

Optimized fixation and immunofluorescence staining methods for Dictyostelium cells.

Monica Hagedorn¹, Eva M Neuhaus, Thierry Soldati

¹Department of Biochemistry, University of Geneva, Switzerland.

Methods in Molecular Biology (Clifton, N.J.)

|September 8, 2006

Summary

Improving fluorescence microscopy requires better cell preservation. This study details modified aldehyde fixation and rapid freezing methods to enhance structural and antigenic integrity for microscopy.

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Techniques for Imaging Prometaphase and Metaphase of Meiosis I in Fixed Drosophila Oocytes

Techniques for Imaging Prometaphase and Metaphase of Meiosis I in Fixed Drosophila Oocytes

Published on: October 31, 2016

Area of Science:

Cell Biology
Microscopy Techniques
Biochemistry

Background:

Advanced fluorescence microscopy demands superior structural and antigenic preservation in fixed cells.
Conventional chemical fixation methods often cause antigenicity loss, membrane damage, and cytosol leakage.
Solvent fixation, while bypassing permeabilization, can lead to significant molecular extraction.

Purpose of the Study:

To present optimized protocols for chemical fixation and immunostaining.
To introduce novel rapid freezing techniques for improved cellular preservation.
To minimize artifacts associated with fixation and permeabilization in microscopy.

Main Methods:

Modification of standard aldehyde fixation protocols.
Development of rapid freezing techniques using various coolants.
Low-temperature solvent fixation following cryo-fixation.
Detailed protocols for light and electron microscopy applications.

Main Results:

Reduced artifacts, such as organelle vesiculation and cytosol leakage, compared to standard methods.
Enhanced antigenicity and structural integrity of cellular components.
Successful preservation of both soluble and membrane-bound molecules.
Compatibility of methods for both light and electron microscopy.

Conclusions:

Modified aldehyde fixation and cryo-fixation/low-temperature solvent fixation offer superior cellular preservation for advanced microscopy.
These techniques mitigate common fixation-induced artifacts, improving data quality.
Optimized protocols are essential for achieving high-resolution imaging in cell biology.