Search Thermo Fisher Scientific
- Contact Us
- Quick Order
-
Don't have an account ? Create Account
Search Thermo Fisher Scientific
Useful as cell delineation reagents for HCS platforms, HCS CellMask Stains label the entire cell (i.e., cytoplasm and the nucleus) to provide a description of a cell’s anatomy, and provide an accurate backdrop against which the features of interest can be assessed. HCS CellMask stains are available in a range of fluorescent colors, they can be applied to cells immediately after fixation or in the last step of multiplexing protocols, and they are compatible with detergent-based permeabilization. This protocol produces sufficient stain for one 96-well plate at a staining volume of 100 μL/well.
1. Add 25 μL DMSO (Component B) to the entire contents of the HCS CellMask Stain (Component A) to make a 10 mg/ mL stock solution. |
2. On the day of the assay, prepare a fixative solution by adding 2.5 mL 16% aqueous paraformaldehyde (PFA) to 7.5 mL PBS, to obtain a 4% PFA solution. |
3. On the day of the assay, prepare a permeabilization solution by adding 10 μL Triton X-100 to 10 mL PBS. |
4. On the day of the assay, prepare a staining solution by adding 2 μL of the HCS CellMask Stain stock solution to 10 mL PBS. |
1. Culture cells in an appropriate medium and vessel for HCA or fluorescence microscopy. |
2. Optional: Add a test compound or drug to the cells, and incubate as desired. |
3. Remove the medium. |
4. Add 100 μL fixative solution (4% PFA) to each well. |
5. Incubate for 15 minutes. |
6. Remove the fixative. |
7. Wash the cells 2–3 times in PBS. |
8. Add 100 μL permeabilization solution (Triton X-100) to each well. |
9. Incubate for 15 minutes. |
10. Remove the permeabilization solution. |
11. Wash the cells 2–3 times in PBS. |
12. Optional: Perform antibody labeling. |
13. Add 100 μL staining solution (2 µg/mL HCS CellMask Stain) to each well. |
14. Incubate for 30 minutes, protected from light. |
15. Wash the cells 2–3 times in PBS. |
16. Image the cells. |
Cells stained with HCS CellMask Blue Stain. Alexa Fluor 488 Click-iT Plus EdU, and Alexa Fluor 647 goat anti–mouse IgG and imaged with the Thermo Scientific CellInsight High-Content System.
HCS CellMask Blue | HCS CellMask Green | HCS CellMask Orange | HCS CellMask Red | HCS CellMask Deep Red | |
---|---|---|---|---|---|
Excitation/Emission (nm) | 346/442 | 493/516 | 556/572 | 588/612 | 650/655 |
Standard filter set | DAPI | FITC | Cy®3 | Cy®3.5 | Cy®5 |
EVOS Light Cube | DAPI | GFP | RFP | Texas Red | Cy®5 |
Storage conditions | ≤–20°C | ≤–20°C | ≤–20°C | ≤–20°C | ≤–20°C |