Dapi Counterstaining

DAPI counterstaining is a fluorescence-based technique used to label cell nuclei and provide spatial context for antibody-based staining in microscopy. DAPI (4′,6-diamidino-2-phenylindole) binds preferentially to adenine-thymine-rich regions of double-stranded DNA; when illuminated with ultraviolet or violet light, it emits blue fluorescence that outlines nuclear morphology. In immunology and infection research, this counterstain helps localize infected or immune cells, assess changes in nuclear structure, and normalize interpretation of immunofluorescence signals across specimens. Because it is rapid and compatible with fixed-cell preparations, DAPI counterstaining supports imaging-based analysis of host-pathogen interactions and tissue responses.

Dapi Counterstaining - Related Videos

Research

JoVE EoE - Caenorhabditis elegans (worm)

Ethanol Fixation and DAPI Staining: A Method to Visualize DNA in C. elegans

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2023

Here, we describe a technique used to visualize genomic DNA in fixed, intact nematodes. The video includes an example protocol to count chromatin in unfertilized C. elegans oocytes.

Cell Death Screening Using DAPI Staining: A Method for Rapid Screening of IR Induced Clonogenic Cell Death in Cancerous Cell Lines

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2023

This video demonstrates the methodology to screen for major cell death profiles in ionizing radiation (IR) induced cancer cell lines using DAPI staining assay. The DAPI-stained nuclei of target cells undergoing cell death exhibit distinct morphologies and can be easily identified by fluorescence microscopy.

Research

JoVE Journal - Genetics
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C. elegans Gonad Dissection and Freeze Crack for Immunofluorescence and DAPI Staining

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Cited by 7 •

2022

This is a commonly used method for C. elegans gonad dissection followed by freeze crack, which produces germline samples for immunofluorescence via antibody staining, or for simple DAPI staining to visualize DNA. This protocol has been successful for undergraduates in a research lab and in a course-based undergraduate research experience.

Detection of Orexin and Cannabinoid Receptor Co-localization in Diet-Induced Obese Zebrafish

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2025

This video demonstrates a protocol for investigating the co-localization of orexin and cannabinoid receptors in normal and diet-induced obesity (DIO) adult zebrafish brain tissue using immunofluorescence.

Immunofluorescence-Based Visualization of DNA Repair Protein Interactions

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2025

In this video, we describe the immunofluorescence method to visualize the localization of the DNA repair proteins γH2AX and 53BP1 at the sites of double-strand DNA breaks in irradiated cell nuclei.

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