Does DAPI work on live cells?

DAPI is generally used to stain fixed cells since the dye is cell impermeant, although the stain will enter live cells when used at higher concentrations. For live-cell staining, Hoechst 33342 dye is a popular cell-permeant nuclear counterstain.

Can you add DAPI to live cells?

Hoechst and DAPI are popular blue fluorescent, nuclear-specific dyes that can be used to stain live or fixed cells. The dyes have minimal fluorescence in solution, but become brightly fluorescent upon binding to DNA. Therefore, they can be used to stain cells without a wash step.

Does DAPI stain live or dead cells?

DAPI is a fluorescent stain that binds strongly to A-T-rich regions in DNA. DAPI and PI only inefficiently pass through an intact cell membrane and, therefore, preferentially stain dead cells.

Is DAPI cell membrane permeable?

Both DAPI and Hoechst are cell permeable. The main difference is that the DAPI is more toxic so if you stain live cells they will not be alive for long.

Does DAPI interfere with PCR?

No evidence of DAPI interference on quantitative PCR and short tandem repeat (STR) typing.

Where does DAPI bind in the nucleus?

It is believed that DAPI associates with the minor groove of double-stranded DNA, with a preference for the adenine-thymine clusters. Cells must be permeabilized and/or fixed for DAPI to enter the cell and to bind DNA.

Why do we use DAPI?

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.

Does DAPI stain cell nuclei?

DAPI stain won’t identify apoptotic cells, it is only a nuclei staining, identify nuclei of all cells. To identify apoptotic cells, use TUNEL stain or Caspase-3 IHC stain.

Why is DAPI used in fluorescence microscopy?

DAPI binds AT-rich DNA preferentially, so that phytoplasmas, localized among phloem cells, can be visualized in a fluorescence microscope. The procedure is quick, easy to use, inexpensive, and can be used as a preliminary or quantitative method to detect or quantify phytoplasma-like bodies in infected plants.

Do you have to Permeabilize for DAPI?

Sample preparation. Use the fixation protocol appropriate for your sample. DAPI staining is normally performed after all other staining. Note that fixation and permeabilization of the sample are not necessary for counterstaining with DAPI.

Is DAPI excited by UV light?

Normally, DAPI bound to DNA is maximally excited by ultraviolet (UV) light at 358 nm, and emits maximally in the blue range, at 461 nm.