Trypan Blue Staining Solution (0.4%) 

Trypan Blue is a hydrophilic tetrasulfonated anionic dye widely used in cell biology for rapid and straightforward assessment of cell membrane integrity and cell viability. The principle of the assay is based on the ability of viable cells to maintain intact plasma membranes, thereby excluding the dye and remaining unstained. In contrast, non-viable or membrane-damaged cells allow the dye to enter the cytoplasm and appear distinctly blue. By counting stained versus unstained cells, cell death rates and overall viability can be accurately quantified.

Trypan Blue–stained samples can be directly analyzed using an automated cell counter or a hemocytometer to obtain reliable viability measurements. In addition, macrophages can phagocytose Trypan Blue due to their innate engulfment capability, making the dye suitable for use as a vital stain for macrophages. It should be noted that apoptotic bodies typically exclude Trypan Blue, and this distinction should be considered during data interpretation.

MCE Trypan Blue Staining Solution (0.4%) is suitable for viability assessment of most adherent and suspension cell lines. It is commonly used for cell culture quality control, evaluation of cellular responses following drug treatment, and viability measurements after cell thawing and recovery.

RT, 1 year.

1. Cell Collection

For adherent cells, detach the cells using trypsin and/or EDTA; suspension cells can be collected directly. Centrifuge at 1000-2000 rpm for 1 min, discard the supernatant, and resuspend the pellet to prepare a single-cell suspension. Dilute the suspension as required for subsequent experiments.

2. Trypan Blue Staining

Mix the cell suspension with 0.4% Trypan Blue solution at a 9:1 volume ratio (final Trypan Blue concentration: 0.04%). Incubate for 3 min. The staining time may be extended appropriately, but should not exceed 10 min.

3. Cell Counting

Load a small amount of the stained cell suspension onto a hemocytometer for counting. Non-viable cells appear blue, swollen, and dull, whereas viable cells remain unstained with intact morphology and a bright appearance. To ensure accuracy, at least 500 cells should be counted for each sample.

Cell Viability (%) = Viable Cell Count / (Viable Cell Count + Non-viable Cell Count) × 100%

1. Staining time should not be excessively long. Prolonged staining may cause viable cells to gradually take up Trypan Blue and exhibit slight coloration, which can lead to an underestimation of cell viability.

2. A small amount of dye precipitation may occur when the product is stored at low temperatures or left at room temperature for an extended period. This is normal. The precipitate can be redissolved by warming in a water bath. If a small amount of insoluble material remains, the solution may be filtered through a membrane filter before use; this does not affect experimental results.

3. This product is supplied sterile, and it is recommended to handle it in a biosafety cabinet to avoid microbial contamination, which may interfere with cell condition and assay accuracy.

4. This product can be used in parallel with cell counting, allowing simultaneous assessment of cell number and viability, thereby improving experimental efficiency and data reliability.

5. This product is for R&D use only, not for drug, household, or other uses.

6. For your safety and health, please wear a lab coat and disposable gloves to operate.