Bacterial Viability and Vitality Assays for Flow Cytometry
 | Optimized for use in bacterial cells, our BacLight™ series of bacterial viability and vitality assays for flow cytometry allow clear separation of living and dead cells based on various cellular characteristics. Due to the inherent differences between prokaryotic and eukaryotic cells, specialized dyes must be used to differentiate live and dead cell populations in bacteria.
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Membrane Integrity Viability Kits
The LIVE/DEAD® BacLight™ Bacterial Viability Kit for flow cytometry is convenient and easy-to-use for monitoring the viability of bacterial populations as a function of the membrane integrity of the cell. Cells with a compromised membrane that are considered to be dead or dying will stain red, whereas cells with an intact membrane will stain green.
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Figure 1. Micrococcus luteus and Bacillus cereus stained with the LIVE/DEAD® BacLight™ Bacterial Viability Kit for flow cytometry. When incubated with the SYTO® 9 and propidium iodide nucleic acid stains provided in this kit, live bacteria with intact cell membranes fluoresce green and dead bacteria with compromised membranes fluoresce red.
| Product | Laser | Ex/Em* | Live Cell Stain | Dead Cell Stain | Cat. No. | |
|---|---|---|---|---|---|---|
| LIVE/DEAD® BacLight™ Bacterial Viability Kit | 488 nm | 480/500 nm (L), 490/635 nm (D) | SYTO®9 | Propidium Iodide | L7012 | Order |
*Indicates excitation and emission. L = live, D = dead.
Publications Where this Product is Referenced
McIver, AM, et al. (2008) Microbial Oxidation of Naphthalene to cis-1,2-Naphthalene Dihydrodiol Using Naphthalene Dioxygenase in Biphasic Media. Biotechnology Progress 24:593-598.
İleri, N., et al. (2007) Phosphate enrichment and fed-batch operation for prolonged β-lactamase production by Bacillus licheniformis. J. of Applied Micro. (102) 1418–1426.
Berney, M., et al. (2007) Assessment and Interpretation of Bacterial Viability by Using the LIVE/DEAD BacLight Kit in Combination with Flow Cytometry. App. Env. Micro., 73:3283-3290.
El-Nezami, H. et al. (2004) Chemical Moieties and Interactions Involved in the Binding of Zearalenone to the Surface of Lactobacillus rhamnosus Strains GG. J. Agric. Food Chem., 52: 4577–4581.
Metabolic Activity Viability Kits
The BacLight™ RedoxSensor™ Bacterial Vitality Assay for flow cytometry provides effective reagents for evaluating bacterial cell health and vitality based on detecting activity of bacterial oxidases and reductases. Enhanced fluorescent signal is observed in bacterial cells with active metabolic processes. The BacLight™ RedoxSensor™ Bacterial Vitality kits also withstand fixation procedures.
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Figure 2. BacLight™ RedoxSensor™ Green Bacterial Vitality Kit. A mixture of healthy and killed Staphylococcus aureus cells were stained with 100 nM RedoxSensor™ Green reagent and propidium iodide. Dual-color flow cytometric analysis of the sample, gated on bacteria using scatter, shows both the healthy and the membrane-compromised cell populations. Similar results can be obtained with Escherichia coli.
| Product | Laser | Ex/Em* | Live Cell Stain | Dead Cell Stain | Cat. No. | |
|---|---|---|---|---|---|---|
| BacLight™ RedoxSensor™ Green Bacteria Vitality Kit | 488 nm | 490/520 nm (L), 490/635 nm (D) | SYTO®9 | Propidium Iodide | B34954 | order |
| BacLight™ RedoxSensor™ CTC Vitality Kit | 488 nm | 480/500 nm (L), 490/635 nm (D) | SYTO®24 | Propidium Iodide | L7012 | order |
*Indicates excitation and emission. L = live, D = dead.
Publications Where this Product is Referenced
Yeom, J. et al. (2010) Iron Homeostasis Affects Antibiotic-mediated Cell Death in Pseudomonas Species. Journ. Biol. Chem. 285:22689-22695.
Lyon, DL and JJ Alvarez (2008) Fullerene Water Suspension (nC60) Exerts Antibacterial Effects via ROS-Independent Protein Oxidation. Envir. Sci. Tech. 42:8127-8132.
Kalyuzhnaya, MG, et al. (2008) Real-time detection of actively metabolizing microbes by redox sensing as applied to methylotroph populations in Lake Washington. ISME Journal 2:696-706.
Bacterial Membrane Potential Kits
The BacLight™ Bacterial Membrane Potential Kit for flow cytometry provides the fluorescent membrane-potential indicator dye, DiOC2(3), which exhibits green fluorescence in all bacterial cells, but it becomes more concentrated in healthy cells that are maintaining a membrane potential, causing the dye to self-associate and the fluorescence emission to shift to red. The red- and green-fluorescent bacterial populations are easily distinguished using a flow cytometer.
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Figure 3. BacLight™ Bacterial Membrane Potential Kit for flow cytometry. The red-versus-green fluorescence dot plot show Staphylococcus aureus incubated with 30 µM DiOC2(3) for 30 minutes in either the presence or absence of 5µM carbonyl cyanide 3-chlorophenylhydrazone (CCCP). Flow cytometer data were collected with log amplification.
| Product | Laser | Ex/Em* | Live Cell Stain | Dead Cell Stain | Cat. No. | |
|---|---|---|---|---|---|---|
| BacLight™ Bacterial Membrane Potential Kit | 488 nm | 482 nm/497 nm | DiOC2(3) | None | B34950 | order |
*Indicates excitation and emission.
Publications Where this Product is Referenced
Leys N, et al. (2009) The response of Cupriavidus metallidurans CH34 to spaceflight in the international space station. Antonie Van Leeuwenhoek 96:227-245.
Lyon, DL and JJ Alvarez (2008) Fullerene Water Suspension (nC60) Exerts Antibacterial Effects via ROS-Independent Protein Oxidation. Envir. Sci. Tech. 42:8127-8132.
Rao, SPS, et al. (2008) The protonmotive force is required for maintaining ATP homeostasis and viability of hypoxic, nonreplicating Mycobacterium tuberculosis. PNAS 105:11945-11950.
