SPHERO Ultra Rainbow Fluorescent Particles
- Consists of a single peak for optical alignment of any flow cytometer on all channels from UV to far-red.
- Determines if the flow cell is clean and has no fluid clogging.
- Measures the coefficients of variation (CV), peak channels and histogram distribution to determine flow functionality cytometers.
New flow cytometers with an increasing number of fluorescent channels in the ultraviolet and far-red rays and availability of corresponding fluorescent conjugates are now available. To keep up to date, we have developed Ultra-rainbow fluorescent particles with enhanced UV rays and the intensity of the far-red fluorescence. The Ultra Rainbow Fluorescent particles contain a single peak and are designed to check the optical alignment of any flow cytometer on all channels.
Instructions for use:
A. Particle preparation
- Shake the particles vigorously.
- Add 2 to 4 drops of particles to 1 ml of pod fluid or deionized water. The inclusion of a small amount of detergent (~ 0.01%) in dilution buffer increases the percentage of the singlet population.
B. Daily alignment
To determine the optical alignment of the system do the following:
- Set an active gate for the singlet population at the FSC vs SSC histogram to exclude aggregates.
- Adjust the gain and high voltage so that the mean channel number of the peak is in a default position in each histogram of interest. The histograms on the previous page can be used as a guide.
- Collect 5000 events inside the door.
- Record% CV and high voltage for all Fluorescence channels of interest.
- Use a computer program such as Excel to generate the Levy Jennings graphs.
NOTE: If the values of any parameter exceed those of the daily or pre-established average values, which are determined by at least one month of data, additional calibration or alignment procedures must be performed in accordance with the instrument’s operating manual.