There are two flow cytometers available:
Flow Cytometer (analyzer) BD LSR II has two lasers:
- 488 nm/ Blue laser SAPPHIRE 20mW
- 638 nm/ Red Laser HeNe 30mW
Cells can be inserted to the machine from tubes and 96-well plates.
Cell Sorter BD FACSAria has three lasers:
- 355 nm / UV Laser XYTE 20 mW
- 488 nm / Blue Laser SAPPHIRE 100 mW
- 639 nm / Red Laser CUBE 40 mW
Cells must be supplied from tube and can be sorted into 4 different tubes or to the 96-well plate.
How to operate flow cytometer
- Book your time if you plan to use the machine. Please be precise, do not book it for the whole day if you use it only for an hour. Respect your colleagues and finish in time. When done, please write down your working time to the notebook just next to the machine.
- Switch the flow cytometer on at least 30 minutes before start, it needs to warm up. It is highly recommended to wash the machine before you start.
- Make sure your dyes mach our lasers and filters. Check out our laser parameters. SpectraViewer and Spectrum Guide can be useful when planning your experiment.
- Please provide 0.3 - 1 ml of cell suspension in the tube. Mix before analysis to make sure there are no cell clumps and cells have not precipitated.
- On 96-well plate please provide 200 - 250 microliters of cell suspension. The maximum amount flow cytometer can take is 200 microliters and there should be something left to avoid air getting into the machine.
- Recommended flow rate is 500 - 700 events/sec from 96-well plate and up to 1700 events/sec. frome tube. If you are not sure about the concentration of your cells in probe start with higher concentration, it is easy to dilute it if necessary.
- Before you start make sure you have proper negative and positive controls. Inexperienced users are encouraged to consult with core facility people and more experienced colleagues, preferentially already during the planning of experiments.
- After finishing your work wash the machine and clear out the spaces. The last user must turn the machine off.
- No food or drinks are allowed! Wear spear shoes.