- Fluorescence detection
- Ready-to-use solution
Chemical Name: 4',6-Diamidino-2-phenylindole, dihydrochloride, solution
CAS: 28718-90-3 (DAPI)
Contents of the kit: DAPI Solution 25 µl x 4 vials
Storage Condition : 0-5ºC, protect from light
Shipping Condition : ambient temperature
Required Equipment and Materials
10 μl, 1000 μl pipettes, incubator, Microscope (blue excitation filter and red emission filter) or flow cytometer (360 nm violet laser)
Staining procedure
1. Allow DAPI solution a) to stand at room temperature for 30 minutes. Solution should be protected from light.
2. Resuspend the organisms with PBS(-) or saline and adjust the number of cells to 106cells/mL(flow cytometry) or 108-109cells/mL(microscopy).
3. Add 1 μL of DAPI solution into the microbial cell suspension and vortex gently to mix. Formaldehyde-fixation can be recommended if necessary.
4. Incubate the microbial cells at room temperature for 5 minutes.
5. Analyze the stained-cells by a flow cytometer or a microscope. The maximum wavelengths of the dye are 360 nm for excitation and 460 nm for emission.
a) Since DAPI may be carcinogenic, be careful when handling and disposing.
Staining Data
Fig. 2 L. casei stained with CTC and DAPI (left). B. cereus stained with CFDA and DAPI (right).