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LIVE/DEAD BacLight Bacterial Viability Kits Manual / Product Insert

  • Version: 07-15-2004
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Manual: LIVE/DEAD BackLight Bacterial Viability Kit for microscopy, microscopy and quantitative assays and 10 applicator sets (Japanese) Manual / Product Insert

  • Version: 15July2004
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When using the LIVE/DEAD BacLight kit for bacterial viability, with SYTO 9 and propidium iodide (PI), why are some cells detected with both dyes? Shouldn't live cells be green and dead cells be red? Product FAQ

Answer

SYTO 9 dye will enter all cells, live or dead. PI only enters cells with compromised membranes (dead cells or damaged cells). First, perform single color staining and examine under both filter sets. Longpass filters or the use of too much dye may result in excessive bleedthrough of the green dye emission into the red channel and the emission of PI in the green channel. Use narrower bandpass filters if possible, and use lower concentrations of the dye. Some live cells may take up PI by engulfment; avoid extended incubation times with the dye. Apply PI to a live cell culture and optimize incubation times to limit engulfment.

Find additional tips, troubleshooting help, and resources within our Cell Analysis Support Center.

Answer Id: E6493

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I want to use propidium iodide and SYTO 9 to do LIVE/DEAD testing of a bacterial sample. Will anaerobic conditions adversely affect the assay? Product FAQ

Answer

Oxygen content should not affect the binding of propidium iodide and SYTO 9 to nucleic acids. SYTO 9 will label all cells, and propidium iodide will label only dead cells or cells with a compromised membrane.

Find additional tips, troubleshooting help, and resources within our Cell Analysis Support Center.

Answer Id: E6462

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I have a LIVE/DEAD BacLight Bacterial Viability kit that has SYTO 9 and propidium iodide in it. Will I be able to stain eukaryotic cells that have engulfed bacteria and determine if the bacteria are alive or dead using this kit? Product FAQ

Answer

Unfortunately, no. SYTO 9 will label the nuclei of live or dead cells, including the eukaryotic cells. Propidium iodide is cell impermeant, and will only enter dead cells. If the eukaryotic cells are dead, they will label with propidium iodide as well. If the eukaryotic cells are alive, propidium iodide will not be able to enter and thus will not label the bacteria inside, whether the bacteria are alive or dead. We are not aware of any way to do a viability assay of bacteria once they have been engulfed by cells.

Find additional tips, troubleshooting help, and resources within our Cell Analysis Support Center.

Answer Id: E6456

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What bacterial parameters can I look at by flow cytometry? Product FAQ

Answer

You can stain bacteria with a general stain such as BacLight Green Bacterial Stain (Cat. No. B35000) or BacLight Red Bacterial Stain (Cat. No. B35001). You can look at gram character (Cat. No. L7005), cell viability (Cat. Nos. L7007, L7012, and L13152), cell count (Cat. Nos. L34856 and B7277), and cell vitality. Cell vitality can be measured by membrane potential (Cat. No. B34950) or by metabolism (Cat. Nos. B34954 and B34956).

Find additional tips, troubleshooting help, and resources within our Cell Analysis Support Center.

Answer Id: E14852

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