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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. [ 7 ] It is labeled non-toxic in its MSDS [ 8 ] and though it was not shown to have mutagenicity to E. coli , [ 9 ] it is labelled as a known mutagen in manufacturer information. [ 2 ]
Key differences between Hoechst dyes and DAPI are: Hoechst dyes are less toxic than DAPI, which ensures a higher viability of stained cells. [5] The additional ethyl group in certain Hoechst dyes (Hoechst 33342) renders them more cell-permeable. [6] There are nuclei staining dyes that allow for viability of cells after staining. [citation needed]
DAPI is a fluorescent nuclear stain, excited by ultraviolet light and showing strong blue fluorescence when bound to DNA. DAPI binds with A=T rich repeats of chromosomes. DAPI is also not visible with regular transmission microscopy. It may be used in living or fixed cells. DAPI-stained cells are especially appropriate for cell counting. [18]
Propidium Iodide is not membrane-permeable, making it useful to differentiate necrotic, apoptotic and healthy cells based on membrane integrity. [3] [4] PI also binds to RNA, necessitating treatment with nucleases to distinguish between RNA and DNA staining. [5] PI is widely used in fluorescence staining and visualization of the plant cell wall ...
FISH has also been successfully done on unfixed cells. [11] After fixation, samples are permeabilized to allow the penetration of hybridization reagents. The use of detergents at a 0.1% concentration is commonly used to enhance the tissue permeability such as Tween-20 or Triton X-100. [12]
Green = smooth muscle actin (SMA) with Alexa 488 fluorophore. Blue = DAPI counterstain. Red = auto-fluorescence. Immunofluorescence (IF) is a light microscopy-based technique that allows detection and localization of a wide variety of target biomolecules within a cell or tissue at a quantitative level.
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Left: DAPI stained female human fibroblast with Barr body (arrow). Right: histone macroH2A1 staining. Arrow points to sex chromatin in DAPI-stained cell nucleus, and to the corresponding sex chromatin site in the histone macroH2A1-staining. A Barr body (named after discoverer Murray Barr) [1] or X-chromatin is an inactive X chromosome.
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