Can Hoechst stain bacteria?
Hoechst and DAPI stain bacteria more dimly than mammalian cells. Live or killed bacteria (gram-negative or gram-positive) can be stained with 12-15 ug/mL Hoechst or DAPI in PBS or 150 mM NaCl for 30 minutes at room temperature.
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.
How does Hoechst bind to DNA?
Hoechst 33258 binds to DNA in two different modes: the high affinity (Kd 1–10 nM) binding results from the specific interaction with B-DNA minor groove and the low affinity (Kd ~1000 nM) reflects the nonspecific interaction with DNA sugar–phosphate backbone [22].
Is Hoechst 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 Hoechst bind to RNA?
3.6 RNA does not interfere significantly with the DNA assay because Hoechst 33258 does not normally bind to RNA.
What is Hoechst method?
Invitrogen Hoechst 33342 nucleic acid stain is a popular cell-permeant nuclear counterstain that emits blue fluorescence when bound to dsDNA. This dye is often used to distinguish condensed pycnotic nuclei in apoptotic cells and for cell cycle studies in combination with BrdU. It is also available as a solution (Cat.
What is the role of Hoechst DNA staining in the diagnosis of mycoplasma infection?
DNA fluorochrome staining with Hoechst 33258 bisbenzimide is commonly used for detection of mycoplasma contamination in cell cultures. Photobleaching of Hoechst 33258 is pronounced under the conditions of intense illumination, high magnification and resolution required for detection of mycoplasmas.
Can DAPI stain non permeabilized cells?
In my experience, DAPI works best with permeabilised cells. Hoechst is weakly permeable and you would get some staining with that. Dear Rosemary, DAPI labelling does not require neither permeabilisation nor fixation.
Does Hoechst stain mycoplasma?
What is Hoechst 33342 stain used for?
Description. 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.
How to label cells with Hoechst stain?
Labeling cells 1 Culture cells in an appropriate medium and vessel for fluorescence microscopy. 2 Prepare the Hoechst staining solution by diluting the Hoechst® stock solution 1:2,000 in PBS. 3 Remove the medium. 4 Add sufficient staining solution to cover the cells. 5 Incubate for 5–10 minutes, protected from light.
How to prepare Hoechst stain for fluorescence microscopy?
1. Culture cells in an appropriate medium and vessel for fluorescence microscopy. 2. Prepare the Hoechst staining solution by diluting the Hoechst® stock solution 1:2,000 in PBS. 3. Remove the medium. 4. Add sufficient staining solution to cover the cells. 5. Incubate for 5–10 minutes, protected from light.
What is h0342 stain used for?
By staining intact cells with the DNA binding, vital, fluorescent dye, Hoechst 33342 (H0342), it is possible to sort viable (clonogenic) cells according to DNA content and thereby obtain cells from the G1 S, and G 2 phases of the cell cycle.