Guidelines

Can DAPI be used for fixed cell staining?

Can DAPI be used for fixed cell staining?

DAPI can be used for fixed cell staining. The concentration of DAPI needed for live cell staining is generally very high; it is rarely used for live cells. It is labeled non-toxic in its MSDS and though it was not shown to have mutagenicity to E. coli, it is labelled as a known mutagen in manufacturer information.

What are the fluorescence properties of DAPI?

Fluorescence properties. When bound to double-stranded DNA DAPI has an absorption maximum at a wavelength of 358 nm ( ultraviolet) and its emission maximum is at 461 nm ( blue ). Therefore, for fluorescence microscopy DAPI is excited with ultraviolet light and is detected through a blue/cyan filter. The emission peak is fairly broad.

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What is the emission peak of DAPI?

When bound to double-stranded DNA DAPI has an absorption maximum at a wavelength of 358 nm (ultraviolet) and its emission maximum is at 461 nm (blue). Therefore, for fluorescence microscopy DAPI is excited with ultraviolet light and is detected through a blue/cyan filter. The emission peak is fairly broad.

What is the use of DAPI in microbiology?

Outside of analytical fluorescence light microscopy DAPI is also popular for labeling of cell cultures to detect the DNA of contaminating Mycoplasma or virus. The labelled Mycoplasma or virus particles in the growth medium fluoresce once stained by DAPI making them easy to detect.

What is the difference between Hoechst stain and DAPI stain?

The Hoechst stains are similar to DAPI in that they are also blue-fluorescent DNA stains which are compatible with both live- and fixed-cell applications, as well as visible using the same equipment filter settings as for DAPI. ^ a bKapuscinski, J. (September 1995).

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What is the blue emission of DAPI stain?

DAPI’s blue emission is convenient for microscopists who wish to use multiple fluorescent stains in a single sample. There is some fluorescence overlap between DAPI and green-fluorescent molecules like fluorescein and green fluorescent protein (GFP) but the effect of this is small.