Hoechst 33342 (Hydrochloride) is part of the Hoechst family of fluorescent stains that also includes Hoechst 33258 and 34580. Hoechst 33342 (Hydrochloride) is a cell-permeable, fluorescent dye that is recommended for staining and visualization of cell nuclei in live or fixed cells. The excitation of Hoechst 33342 (Hydrochloride) occurs around an ultraviolet wavelength of 350 nm, with maximal emission occurring at 461 nm, emitting a blue/cyan fluorescent light. Compared to the other Hoechst stains, Hoechst 33452 is more readily cell-permeable due to the presence of an additional lipophilic ethyl group (Bucevičius et al.). For live cultures, Hoechst 33342 (Hydrochloride) is recommended to be used in place of DAPI, as it is less toxic and more capable of preserving cell viability after staining. Hoechst 33342 (Hydrochloride) may also be used as a nuclear counterstain in fixed samples.
Data Figures
Figure 1. Hoechst 33342 Labels DNA in Live Cell Cultures
(A) Live HeLa cells were stained with 5 µM Hoechst 33342 (Hydrochloride) and then imaged using a fluorescence microscope. Image was taken at 200X magnification and is a cropped field of view. The image shows bright staining of several HeLa cell nuclei (blue). (B) Chemical structure of Hoechst 33342 (Hydrochloride).
Figure 2. Hoechst 33342 (Hydrochloride) Brightly Labels DNA in Cell Cultures Pre-Fixation and Post-Fixation
Live HeLa cells were stained with (A) 5 µM or (B) 10 µM Hoechst 33342 (Hydrochloride; blue) for 30 minutes at 37°C, then imaged using a fluorescence microscope. (C, D) Images were taken at 200X magnification and are cropped field of views. The images show bright staining of DNA is maintained even after fixation with 4% formaldehyde.
Protocols and Documentation
Find supporting information and directions for use in the Product Information Sheet or explore additional protocols below.
Document Type
Product Name
Catalog #
Lot #
Language
Document Type
Product Information Sheet
Product Name
Hoechst 33342 (Hydrochloride)
Catalog #
100-1540
Lot #
All
Language
English
Document Type
Safety Data Sheet
Product Name
Hoechst 33342 (Hydrochloride)
Catalog #
100-1540
Lot #
All
Language
English
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