Hoechst 33342 Fluorescent Nucleic Acid Stain

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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 pyknotic nuclei in apoptotic cells and for cell cycle studies.

Figure 1. MR-(DEVD)2 (cat. 936) and Hoechst 33342 were used to quantify apoptosis in human PASMCs. Cells were treated with a negative control (A), a drug that inhibits proliferation (B), or H2O2 to induce apoptosis (C). Apoptotic cells stained red with cleaved MR-(DEVD)2 and blue with Hoechst. They have less intense blue nuclei than healthy cells, which have bright blue nuclei. Research of Dr. F. Perros, et al., Universite Paris-Sud 11; Clamart; Hopital Henri-Mondor, Creteil, France.
Figure 2. THP-1 cells were stained with Poly (active) Caspase FAM FLICA® Inhibitor Reagent (green), and Hoechst 33342 (Blue). Cells were incubated with staurosporine to induce apoptosis. Two photos were taken and superimposed. Only one cell of the three cells is apoptotic (middle) – it is stained positive for caspase activity with FAM-VAD-FMK. Data courtesy of Dr. Brian W. Lee.
Figure 3. MCF-7 cells were exposed to 0.15 µM camptothecin (6210) for 24 hours at 37°C, then stained with MR-(DEVD)2 for 30 minutes at 37°C, washed, and then stained with 1 µg/mL Hoechst stain. Apoptotic cells with orange-red lysosomal bodies and less intense blue nuclei are intermixed with non-apoptotic cells bearing bright blue nuclei and absent or reduced orange-red lysosomal staining. Photo provided by Dr. Zbigniew Darzynkiewicz at Brander Cancer Research Center Institute, New York, NY.
Figure 4. Cell death in primary rat hippocampal neurons. (D) is a composite of FLICA® FAM-DEVD-FMK (B), PI (C), and Hoechst (D). DNA is labeled in 4 cells (A), 3 cells are caspase-positive (B), 2 are membrane-compromised (C). One cell is in early apoptosis (green but not red). Two FLICA®-positive cells are also PI-positive (red, C) (becoming membrane compromised) and are in the late stages of apoptosis rather than necrosis. Data courtesy of Dr. Z. Kahraman Akozer, University of Maryland.
Representative images of BT-20 cells with vehicle (0 μM), 2, or 5 μM of TPH104c for 24 h or paclitaxel (PTX, 1 μM, a positive control) and stained with Hoechst 33342 dye (Cat no 639). TPH104c did not produce condensed or fragmented nuclei compared to cells incubated with paclitaxel (PTX). Scale bar = 25 μM. Image from publication, CC-BY-4.0, PMID: 39123351.
Figure 1. MR-(DEVD)2 (cat. 936) and Hoechst 33342 were used to quantify apoptosis in human PASMCs. Cells were treated with a negative control (A), a drug that inhibits proliferation (B), or H2O2 to induce apoptosis (C). Apoptotic cells stained red with cleaved MR-(DEVD)2 and blue with Hoechst. They have less intense blue nuclei than healthy cells, which have bright blue nuclei. Research of Dr. F. Perros, et al., Universite Paris-Sud 11; Clamart; Hopital Henri-Mondor, Creteil, France.
Figure 1. MR-(DEVD)2 (cat. 936) and Hoechst 33342 were used to quantify apoptosis in human PASMCs. Cells were treated with a negative control (A), a drug that inhibits proliferation (B), or H2O2 to induce apoptosis (C). Apoptotic cells stained red with cleaved MR-(DEVD)2 and blue with Hoechst. They have less intense blue nuclei than healthy cells, which have bright blue nuclei. Research of Dr. F. Perros, et al., Universite Paris-Sud 11; Clamart; Hopital Henri-Mondor, Creteil, France.
Figure 2. THP-1 cells were stained with Poly (active) Caspase FAM FLICA® Inhibitor Reagent (green), and Hoechst 33342 (Blue). Cells were incubated with staurosporine to induce apoptosis. Two photos were taken and superimposed. Only one cell of the three cells is apoptotic (middle) – it is stained positive for caspase activity with FAM-VAD-FMK. Data courtesy of Dr. Brian W. Lee.
Figure 3. MCF-7 cells were exposed to 0.15 µM camptothecin (6210) for 24 hours at 37°C, then stained with MR-(DEVD)2 for 30 minutes at 37°C, washed, and then stained with 1 µg/mL Hoechst stain. Apoptotic cells with orange-red lysosomal bodies and less intense blue nuclei are intermixed with non-apoptotic cells bearing bright blue nuclei and absent or reduced orange-red lysosomal staining. Photo provided by Dr. Zbigniew Darzynkiewicz at Brander Cancer Research Center Institute, New York, NY.
Figure 4. Cell death in primary rat hippocampal neurons. (D) is a composite of FLICA® FAM-DEVD-FMK (B), PI (C), and Hoechst (D). DNA is labeled in 4 cells (A), 3 cells are caspase-positive (B), 2 are membrane-compromised (C). One cell is in early apoptosis (green but not red). Two FLICA®-positive cells are also PI-positive (red, C) (becoming membrane compromised) and are in the late stages of apoptosis rather than necrosis. Data courtesy of Dr. Z. Kahraman Akozer, University of Maryland.
Representative images of BT-20 cells with vehicle (0 μM), 2, or 5 μM of TPH104c for 24 h or paclitaxel (PTX, 1 μM, a positive control) and stained with Hoechst 33342 dye (Cat no 639). TPH104c did not produce condensed or fragmented nuclei compared to cells incubated with paclitaxel (PTX). Scale bar = 25 μM. Image from publication, CC-BY-4.0, PMID: 39123351.

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Data Sheet Safety Data Sheet

SKU: 639

Size: 1 mL
1-2 business days
Price:
$22.75
Hoechst 33342 is a popular cell-permeant, blue fluorescent nuclear stain. It is used to visualize the nuclei of living or fixed cells and tissues and is often used to distinguish condensed, pyknotic nuclei in apoptotic cells. Hoechst 33342 emits blue fluorescence when bound to double stranded DNA. It is slightly more membrane permeant than the Hoechst 33258 analog. Hoechst 33342 may be used to identify healthy or apoptotic nuclear morphology and for cell cycle studies.
350 nm / 480 nm
Flow cytometry, Fluorescence microscope
2-8°C
Domestic: Overnight Delivery; International: Priority Shipping
5.0 + 0.5
200 µg/mL
United States
  1. Add Hoechst 33342 to the cell sample media at 0.5% v/v. For example, add 1.5 µL Hoechst to 300 µL of cells. 2. Incubate 10-20 minutes at room temperature.
  2. Visualize with a fluorescence microscope by using a UV excitation and blue emission filter. The blue Hoechst stain fluoresces at 461 nm.
  3. Alternatively, cells may be analyzed with a flow cytometer using a UV excitation source.
  4. When bound to dsDNA, the maximum absorption is 350 nm and the maximum emission is 461.

Product Specific References

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