What is the excitation and emission for DAPI?

DAPI (4,6-Diamidino-2-phenylindole) is a fluorescent compound with an excitation peak at 359 nm and an emission peak at 457 nm, giving it a fairly large Stokes’ Shift of 98nm.

What is DAPI excitation?

DAPI (4′,6-diamidino-2-phenylindole) is a blue-fluorescent DNA stain that exhibits ~20-fold enhancement of fluorescence upon binding to AT regions of dsDNA. It is excited by the violet (405 nm) laser line and is commonly used as a nuclear counterstain in fluorescence microscopy, flow cytometry, and chromosome staining.

What is DAPI emission?

DAPI – An easy-to-use Nuclear Dye DAPI (4′,6-diamidino-2-phenylindole) is a well-characterized blue-emitting fluorescent compound widely used for nuclear staining. DAPI is a DNA minor groove binding compound which can permeate cell membranes therefore preferentially stains nuclei (Figure 1).

Does DAPI only emit UV light?

The fluorescent dye 4′-6-Diamidino-2-phenylindole (DAPI) is frequently used in fluorescence microscopy as a chromosome and nuclear stain because of its high specificity for DNA. 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.

What is the excitation wavelength of DAPI?

Nuclei in the tissue section were targeted with the nucleic acid probe DAPI, which has an excitation maximum at 358 nanometers and an emission maximum at 461 nanometers when bound to DNA in cell cultures and tissue sections.

How does DAPI enter the cell?

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. Fluorescence increases approximately 20-fold when DAPI is bound to double-stranded DNA.

How does DAPI staining work?

As DAPI can pass through an intact cell membrane, it can be used to stain both live and fixed cells, though it passes through the membrane less efficiently in live cells and therefore provides a marker for membrane viability….DAPI.

What is DAPI used for in flow cytometry?

Therefore, DAPI is used for evaluation of cell death and apoptosis of unfixed cells in flow cytometry. Additionally, DAPI may be used as a nuclear counterstain of fixed cells in imaging or flow cytometry or for determination of DNA content in cell cycle analysis.

How 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.

Does DAPI stain apoptotic cells?

Additionally, apoptotic cells stained with DAPI may have observable nuclear blebbing which may help in differentiating necrotic cells which do not blebb.

How does DAPI specifically label mitotic cell DNA?

DAPI specifically stains double stranded DNA with no non-specific labeling in the cytoplasm. For DAPI to enter cells and bind to DNA, the cells need to be permeabilized. After binding to DNA, the fluorescence of DAPI increases 20 fold.

Why is DAPI used in fluorescence microscopy?