I am using the Flow Cytometry Sub-micron Particle Size Reference Kit and can't resolve the 0.1 and 0.2 µm beads in my flow run. What can I do to get good separation?
We recommend mixing the beads thoroughly before use and using a slower flow rate.
Find additional tips, troubleshooting help, and resources within our Cell Analysis Support Center.
Are there any surface modifications of the beads within the Flow Cytometry Sub-micron Particle Size Reference Kit?
Yes. Charged groups are on the surface making the beads water-dispersible.
Find additional tips, troubleshooting help, and resources within our Cell Analysis Support Center.
Where is the dye localized in the beads within the Flow Cytometry Sub-micron Particle Size Reference Kit?
The dye is dispersed throughout the polystyrene matrix.
Find additional tips, troubleshooting help, and resources within our Cell Analysis Support Center.
When using the Live/Dead BacLight Bacterial Viability and Counting Kit, for flow cytometry, some cells seem to have both red and green signal. Are these cells dead or alive or dying?
The green dye in the kit will label all the cells as it is a cell-permeant nucleic acid stain. The red dye is not cell permeant, and will only stain the cells with compromised membranes (dead cells). Therefore, any cells with red signal will be considered dead. It is possible that you will have some cells that are only red, some that are red and green, and some that are only green. Sometimes the red dye will displace the green dye. In any case, any red cells are dead.
Also, the green dye emission may bleed through into the red channel. Do a single-color staining and examine under both green and red filter sets to determine the level of bleedthrough. To avoid this bleedthrough, use a lower concentration of dye, and, if possible, use narrow bandpass filters.
Find additional tips, troubleshooting help, and resources within our Cell Analysis Support Center.
How can I count my bacteria by flow cytometry?
There are several options. We have two fluorescence based kits that are useful for bacterial counting: Live/Dead BacLight Bacterial Viability and Counting Kit, for flow cytometry (Cat. No. L34856) and Bacteria Counting kit, for flow cytometry (Cat. No. B7277). Another option is the Flow Cytometry Sub-micron Particle Size Reference Kit (Cat. No. F13839).
Find additional tips, troubleshooting help, and resources within our Cell Analysis Support Center.
