Dead cells tend to stain more brightly than live cells. In S. cerevisiae, DAPI and Hoechst preferentially stain dead cells with nuclear and cytoplasmic localization.
Why does Hoechst stain dead cells?
The DNA becomes condensed during apoptosis but not during necrosis. Because of its ability to bind to DNA, Hoechst dyes can be used to observe nuclear condensation, thereby distinguishing apoptotic cells from healthy cells as well as necrotic cells.6 May 2021
What is the difference between Hoechst and DAPI?
Hoechst dyes are typically used for staining DNA content in live cells due to its high cell membrane permeability. DAPI is typically used for staining DNA content in fixed cells due to its low membrane permeability.24 Jul 2018
What color does Hoechst stain?
blue
What is Hoechst staining used for?
Hoechst 33342 is used for specifically staining the nuclei of living or fixed cells and tissues. This stain is commonly used in combination with 5-bromo-2'-deoxyuridine (BrdU) labeling to distinguish the compact chromatin of apoptotic nuclei, to identify replicating cells and to sort cells based on their DNA content.
Can Hoechst stain dead cells?
Since Hoechst can readily cross cell membranes to stain DNA of living and dead cells, it was used to label the total number of cells. In contrast, PI only enters cells with compromised plasma membranes, thus selectively labeling dead cells.12 Sept 2011
Does DAPI stain live or dead cells?
DAPI and PI only inefficiently pass through an intact cell membrane and, therefore, preferentially stain dead cells. TMRM is a cell-permeable fluorescent dye that is sequestered to active mitochondria, and hence labels live cells.1 Apr 2016
What is DAPI and why is it used?
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. 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.
Does Hoechst stain live or dead cells?
Yes, Hoechst 33342 can stain dead cells, however Hoechst 33358 is the preferred dye that's used for staining dead or fixed cells. Because of its ability to bind to DNA, Hoechst dyes can be used to observe nuclear condensation, thereby distinguishing apoptotic cells from healthy cells as well as necrotic cells.6 May 2021
What does Hoechst stain?
Hoechst are non-intercalating dyes that bind the DNA minor groove at A–T-rich regions (Figure 1b). Hoechst dyes stain the DNA of both living and fixed cells and are compatible with immunohistochemistry applications.
Can DAPI stain 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.1 Apr 2016
What does a Hoechst stain show?
Hoechst dyes are commonly used to stain genomic DNA in the following applications: Fluorescence microscopy and immunohistochemistry, often with other fluorophores. Flow cytometry to count or sort out cells.