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DAPI as a useful stain for nuclear quantitation
B I Tarnowski1, F G Spinale, J H Nicholson
1Department of Pathology, Medical University of South Carolina, Charleston 29425.
Abstract:
A simple-to-use fluorescent stain, 4',6-diamidino-2-phenylindole (DAPI), visualizes nuclear DNA in both living and fixed cells. DAPI staining was used to determine the number of nuclei and to assess gross cell morphology. Following light microscopic analyses, the stained cells were processed for electron microscopy. Cells stained with DAPI showed no ultrastructural changes compared to the appearance of cells not stained with DAPI. DAPI staining allows multiple use of cells eliminating the need for duplicate samples.
Insights
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Area of Science:
- Cell Biology
- Microscopy
Background:
- Nuclear DNA visualization is crucial for cell studies.
- Accurate cell counting and morphology assessment are vital.
Purpose of the Study:
- To evaluate 4',6-diamidino-2-phenylindole (DAPI) as a nuclear stain for light and electron microscopy.
- To assess the impact of DAPI staining on cellular ultrastructure and its utility for multiple analyses.
Main Methods:
- Cells were stained with DAPI for nuclear visualization.
- Light microscopy was used to count nuclei and assess cell morphology.
- Stained cells were subsequently processed for electron microscopy to examine ultrastructure.
Main Results:
- DAPI effectively stained nuclear DNA in both living and fixed cells.
- DAPI staining allowed for accurate nucleus counting and gross cell morphology assessment.
- No ultrastructural alterations were observed in DAPI-stained cells compared to unstained controls.
Conclusions:
- DAPI is a reliable and simple fluorescent stain for nuclear DNA.
- DAPI staining does not induce ultrastructural damage, enabling sequential light and electron microscopy.
- This method permits multiple analyses on the same cell sample, reducing the need for duplicates.