Nuclear stain for HCA/HCS cell demarcation
The HCS NuclearMask Blue Stain is used to measure DNA content and perform cell demarcation in both live and fixed cells on high-content imaging and analysis (HCS) platforms. The versatile HCS NuclearMask Stains survive standard formaldehyde-based fixation and detergent-based permeabilization methods. This protocol produces sufficient stain for one 96-well plate at a staining volume of 100 μL/well.
This protocol can be used for:
- Nuclear demarcation in high-content analysis/screening (HCA/HCS)
This protocol should not be used for:
- Flow cytometry
You will need the following for this protocol:
- Cells growing in culture
- HCS NuclearMask Blue Stain (Cat. No. H10325)
- Flat-bottom 96-well microplates
- Optional: Paraformaldehyde, 16% aqueous solution
- Phosphate-buffered saline (PBS)
- HCA instrument or fluorescence microscope
Protocol
Labeling live cells
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Labeling fixed cells
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Spectral information and storage
HCS NuclearMask Blue | |
---|---|
Excitation/Emission (nm) | 350/461 |
Standard filter set | DAPI |
EVOS Light Cube | DAPI |
Storage conditions | 2–6°C |
Protocol tips
- Warm to room temperature and briefly centrifuge the DMSO solution to the bottom of the vial each time before use.
- Treat all nucleic acid binding dyes as potential mutagens and handled with care.
- This stain is supplied as a 2,000X concentrate in DMSO.
Cells stained with HCS NuclearMask Blue Stain and imaged with the Thermo Scientific CellInsight High-Content System.