Is DAPI a good live-cell nuclear label?
DAPI is considered a semi-permeant/impermeant nucleic acid stain. Staining of nucleic is dependent upon the cell line in its performance. Some cell lines will label with DAPI, others not at all, and others label inconsistenly. Instead, we recommend using either Hoechst 33342 or Hoechst 33258, which have the same wavelength and binding mode as DAPI (at the A-T minor groove) but are readily cell-permeant.
Find additional tips, troubleshooting help, and resources within our Cell Analysis Support Center.
I want to label the nuclei of live cells and track them over time. Can I use DAPI for this?
We do not recommend doing this. DAPI is considered to be a semi-permeant/impermeant nucleic acid stain. DAPI staining of live cells may be inconsistent. It is best used as a counterstain for fixed samples. Other cell permeable nucleic acid stains, such as Hoechst or the SYTO dyes may affect cellular function.
For mammalian cells, we recommend using the CellLight Nucleus transduction reagents, available in CFP, GFP and RFP. With these reagents, the cells are transduced overnight in a single labeling step and the next day the nuclei will fluoresce. The label may be retained for 3-5 days and should not affect cell function. Cytoplasmic cell tracking dyes such as the CellTracker dyes may also be used.
Find additional tips, troubleshooting help, and resources within our Cell Analysis Support Center.
I would like to label only 100 µL of sample of the same cell density using a Ready Flow product. The manual notes 2 drops per 1 mL (1 X 10E6 cells/mL). May I scale down for smaller volumes?
Yes, you may scale up or down as needed, but we recommend keeping the cell density the same. There are 41 µL per drop, so 2 drops is 82 µL (for the 1 mL of sample). You need to only scale down for this 100 µL volume, which would be 8.2 µL.
Find additional tips, troubleshooting help, and resources within our Cell Analysis Support Center.
What is the concentration of dye/reagent in the Ready Flow products?
The amount of dye or reagent in the Ready Flow products is proprietary. If you need to consider the exact amount of dye or reagent for your experiment, each of the Ready Flow reagents is available as a standalone product.
Find additional tips, troubleshooting help, and resources within our Cell Analysis Support Center.
Can I use the ReadyProbes reagents for flow cytometry?
This is not recommended. The ReadyProbes reagents were developed for imaging applications whereas the Ready Flow reagents were optimized for flow cytometry.
Find additional tips, troubleshooting help, and resources within our Cell Analysis Support Center.
