SYTOX® Green Nucleic Acid Stain Protocol
Dead cell indicator for microscopy
SYTOX® Green Nucleic Acid Stain is a bright, high-affinity nucleic acid stain that easily penetrates cells with compromised plasma membranes but does not cross the membranes of live cells, making it a useful indicator of dead cells within a population.
This protocol can be used for:
- Nucleic acid (nuclear) staining in fluorescence microscopy
This protocol should not be used for:
- Flow cytometry
You will need the following for this protocol:
- Cells growing in culture
- SYTOX® Green Nucleic Acid Stain (Cat. No. S7020)
- Phosphate-free buffer
- Fluorescence microscope
Labeling fixed cells
First, fix and permeabilize cultured cells with a protocol appropriate for your sample.
1. Wash the cells 1–3 times in a phosphate-free buffer as needed.
|2. Prepare the SYTOX® Green staining solution by diluting the stock solution 1:30,000 (167 nM) in a phosphate-free buffer.|
|3. Add sufficient staining solution to cover the cells.|
|4. Incubate for 15–30 minutes, protected from light.|
|5. Remove the staining solution.|
|6. Wash the cells 2–3 times in a phosphate-free buffer|
|7. Image the cells.|
|Standard filter set||GFP|
|EVOS® Light Cube||GFP|
- Warm to room temperature and briefly centrifuge the DMSO solution to the bottom of the vial each time before use.
- Try multiple dye concentrations in the range from 10 nM to 1 µM to determine the optimal concentration.
- In general, the best results are obtained in buffers that do not contain phosphate, such as Hank’s Balanced Salt Solution (Cat. No. 14025-092).
- Treat all nucleic acid binding dyes as potential mutagens and handle with care.
For Research Use Only. Not for use in diagnostic procedures.