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| Product Name | Bacteria Live/Dead Cell Viability and Cytotoxicity Assay Kit |
| Catalog No. | RESK-0164 |
| Description | This kit contains two fluorescent nucleic acid dyes, DMAO and EthD-III, which stain live bacteria green and dead bacteria red, respectively. DMAO is a green fluorescent nucleic acid dye that stains both live and dead bacteria, while EthD-III is a red fluorescent nucleic acid dye that stains only dead bacteria with compromised cell membranes. When DMAO and EthD-III are used in combination, bacteria with intact cell membranes fluoresce green, whereas bacteria with damaged membranes fluoresce both green and red. Results can be analyzed by fluorescence microscopy or flow cytometry, and the assay is suitable for most bacterial species. Spectral properties: DMAO, Ex/Em = 503/530 nm (with DNA); EthD-III, Ex/Em = 530/620 nm (with DNA). |
| Intended Use | For the differentiation and analysis of live and dead bacteria in viability and cytotoxicity studies; suitable for most bacterial types. For research use only. |
| Principle / Technology | Dual fluorescent nucleic acid staining. DMAO is a membrane-permeant green nucleic acid dye that labels DNA of both live and dead bacteria. EthD-III is a membrane-impermeant red DNA-intercalating dye with AT base preference that enters only bacteria with damaged cell membranes. Bacteria with intact membranes appear green, while membrane-compromised (dead) bacteria appear green and red. |
| Detection Method | Fluorescence microscopy or flow cytometry |
| Sample Type | Bacterial suspensions; suitable for most bacterial types, including bacteria forming biofilms on confocal dishes |
| Performance Range / Specifications | High accuracy, showing good concordance with growth assays performed in liquid or on solid media. DMAO: Ex/Em = 503/530 nm (with DNA) EthD-III: Ex/Em = 530/620 nm (with DNA) |
| Specificity | DMAO is a membrane-permeant nucleic acid dye that stains the DNA of live and dead bacteria; its DNA-binding mechanism is not fully defined. EthD-III is membrane-impermeant and stains only the DNA of dead bacteria; it is a DNA-intercalating dye with AT base preference. Neither dye is affected by esterases. |
| Reaction Conditions / Protocol | 1. Prepare the bacterial suspension: centrifuge to remove the culture medium, collect the bacteria, and wash the cells 3 times with 0.85% NaCl solution. Wash thoroughly to remove residual medium, which would impair staining; do not wash with phosphate buffers such as PBS, as this affects staining. 2. Resuspend the cells in 0.85% NaCl solution. 3. Stain with the DMAO/EthD-III dye mixture and analyze by fluorescence microscopy or flow cytometry. |
| Components / Formulation | DMAO (green fluorescent nucleic acid dye) EthD-III (red fluorescent nucleic acid dye) |
| Storage Conditions | Store at 4 °C. |
| Shelf Life | Stable for at least 6 months when stored at 4 °C. |
| Package Specifications | 100 T |
| Product Form | Kit (liquid dye reagents) |
| Shipping Conditions | Shipped on wet ice. |
| Expiration Date / Stability | Stable for at least 6 months at 4 °C. |
| Regulatory / Compliance | For research use only. |
| Recommended Buffer System | 0.85% NaCl solution for cell washing and suspension; do not use phosphate buffers such as PBS. |
| Application Notes / Precautions | A common criterion of bacterial viability is the ability to reproduce in suitable nutrient medium (growth assay). This kit generally shows good concordance with growth assay results in liquid or on solid media. Under certain conditions, membrane-damaged bacteria may recover and reproduce in nutrient medium but may be scored as dead in this assay; conversely, some bacteria with intact membranes may fail to reproduce but may be scored as live. Consider these possibilities if a considerable discrepancy is observed between this assay and a bacterial growth assay. Depending on the microscope filter set (long-pass vs. short-pass filters), both colors may not be visible simultaneously under a single excitation; if so, excite and observe the two channels separately. In practice, red/green fluorescence co-localization or separate red (dead) and green (live/unhealthy) populations can both be observed; both are normal results, owing to dye-DNA binding behavior and Brownian motion of bacteria in the staining solution. When staining bacteria forming biofilms on confocal dishes, the staining solution may be removed and a PBS wash is optional. |
For research use only, not for clinical use.
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