Trypan Blue Staining Cell Viability Assay Kit is developed based on the principle that healthy intact cells can exclude trypan blue and remain unstained, while cells with lost cell membrane integrity can be stained by trypan blue.Strictly speaking, trypan blue staining detects cell membrane integrity. Generally, cells are considered dead once their membrane integrity is compromised.After trypan blue staining, cell viability can be calculated relatively accurately by direct counting under a microscope or counting after photographing under a microscope.
This kit mainly consists of Trypan Blue stain and Cell Resuspension Solution.The 1 mL reagent package can be used for approximately 10 staining tests.This kit is for research use only and is not suitable for clinical diagnosis or other applications.
Components
| T1509532 | Component | 100T | Storage | | T1509532A | Trypan Blue Stain (2×) | 10 mL | 2-8℃. Store in the dark. | | T1509532B | Cell Resuspension Solution
| 100 mL | 2-8℃. |
|
Materials Provided by User
1. Trypsin digestion solution
2. Microscope, hemocytometer
Protocol (For Reference Only)
1. Cell Collection: For adherent cells, digest with trypsin before collection. Suspension cells can be collected directly.Centrifuge the collected cells at 1000–2000×g for 1 min, discard the supernatant, and resuspend the cell pellet in 1 mL or an appropriate volume of Reagent B.
2. Trypan Blue Staining: Transfer 100 μL of resuspended cells into a standard 1.5 mL or 0.5 mL centrifuge tube.Add 100 μL of Trypan Blue Stain (2×), mix gently, and incubate for 3 min (staining time may be slightly extended but should not exceed 10 min).
3. Counting: Pipette a small amount of stained cells and count using a hemocytometer.For accurate quantification, at least 500 cells per sample are recommended. Count both blue-stained cells and total cells.
Calculation
Cell Viability=(Total Cells-Blue stained Cells)/Total Cells×100%
Precautions
1. Perform aseptic operations in a laminar flow hood to avoid bacterial contamination.
2. Trypan blue is harmful to humans; handle with care.
3. Use the reagents promptly after opening to avoid affecting experimental results.
4. For your safety and health, wear a lab coat and disposable gloves during operation.
