LIVE/DEAD™ BacLight™ Bacterial Viability Kit, 10 Applicator Sets kit - FAQs

View additional product information for LIVE/DEAD™ BacLight™ Bacterial Viability Kits - FAQs (L7007, L13152, L7012)

9 product FAQs found

我如何为LIVE/DEAD细胞活力实验准备死细胞对照？

有两种简单的方法。一种是通过60°C放置20分钟热灭活细胞。第二种是将细胞放入70%乙醇。乙醇固定的细胞可以在冰箱中无限期储存直到使用，可达好几年。

1.离心细胞、使细胞沉淀并去除上清
2.固定细胞：在15mL含有细胞团的试管中加入10 mL70%冰乙醇，先开始逐滴加入后震荡，混合均匀。
3.在冰箱中储存直到使用。
4.快使用时，水洗两次且在选择的缓冲液中重悬。

我可以在流式细胞仪上看到哪些细菌参数?

您可以用BacLight Green细菌染色剂（货号B35000）或BacLight Red细菌染色剂（货号B35001）等常规染色剂进行细菌染色。你们可以看到革兰氏特性（货号L7005），细胞存活率（货号L7007、L7012和L13152），细胞计数（货号L34856和B7277）和细胞活力。细胞活力可以通过膜电位法（货号B34950）或代谢法（货号B34954和B34956）测定。

我可通过流式细胞仪观察细菌和酵母么？

可以。流式细胞仪可用于：

•计数检测：用于流式细胞仪的细菌计数试剂盒（货号B7277）或LIVE/DEAD BacLight细菌活性和计数试剂盒（货号L34856）
•活性/活力检测：LIVE/DEAD BacLight 细菌活性试剂盒（货号 L13152）
•膜电位：BacLight细菌膜电位试剂盒（货号B34950）
•酵母活性/活力实验：用于流式细胞仪（货号L34952）的LIVE/DEAD FungaLight酵母活性试剂盒，FungaLight酵母CFDA, AM/碘化丙啶活力试剂盒（货号F34953）。

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?

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.

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?

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.

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?

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.

How do I prepare dead cell controls for LIVE/DEAD cell viability assays?

There are two easy options. One is to heat-inactivate the cells by placing at 60 degrees C for 20 minutes. The second is to subject the cells to 70% ethanol. Alcohol-fixed cells can be stored indefinitely in the freezer until use, potentially up to several years.

Centrifuge cells, pellet, and remove supernatant.
Fix cells: Add 10 mL ice cold 70% ETOH to a 15 mL tube containing the cell pellet, adding dropwise at first while vortexing, mix well.
Store in freezer until use.
When ready to use, wash twice and resuspend in buffer of choice.

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

What bacterial parameters can I look at by flow cytometry?

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.

Can I look at bacteria and yeast on a flow cytometer?

Yes, you can. We offer:

-Counting assays: Bacteria Counting Kit, for flow cytometry (Cat. No. B7277) or LIVE/DEAD BacLight Bacterial Viability and Counting Kit, for flow cytometry (Cat. No. L34856).
-Viability/vitality assays: LIVE/DEAD BacLight Bacterial Viability Kit (Cat. No. L13152).
-Membrane potential: BacLight Bacterial Membrane Potential Kit (Cat. No. B34950).
-Yeast viability/vitality assays: LIVE/DEAD FungaLight Yeast Viability Kit, for flow cytometry (Cat. No. L34952), FungaLight Yeast CFDA, AM/Propidium Iodide Vitality Kit (Cat. No. F34953)

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