SPHEROT Ultra Rainbow Fluorescent Particles
New movement cytometers, with fluorescent channels from the UV to Far Pink, and corresponding fluorescent conjugates
at the moment are accessible. In consequence, we now have developed the Extremely Rainbow Fluorescent Particles with enhanced UV
and Far Pink fluorescence depth. The Extremely Rainbow Fluorescent Particles include a single peak and are
designed for checking the optical alignment of any movement cytometer in all channels.
Spherotech Cat. No. URFP-30-2 Directions to be used:
A. Preparation of Particles
1. Vortex the particles vigorously
2. Add 2 to Four drops of particles to 1mL of sheath fluid or DI water. The inclusion of a small quantity of detergent (~0.01%) within the dilution buffer will improve the proportion of the singlet inhabitants.
B. Each day Alignment
To find out the optical alignment of the system carry out the next:
1. Set a stay gate for the singlet inhabitants on the FSC vs SSC histogram to exclude aggregates
2. Modify the Acquire and Excessive voltage in order that the imply channel variety of the height is in a predetermined place on every histogram of curiosity. The histograms on the earlier web page can be utilized as a information.
3. Accumulate 5000 occasions contained in the gate
4. Report the % CV and Excessive Voltage for all fluorescence channels of curiosity.
5. Use a pc program resembling Excel to generate the Levy Jennings graphs.
NOTE: If the values on any parameter exceed these of the day-to-day common or preset values, that are decided by not less than one months value of information, extra calibration or alignment procedures must be carried out in response to the instrument operation handbook.
Movement Cytometry Alignment Particles
Spherotech gives all kinds of particles for figuring out if a movement cytometer is correctly aligned and has a clear movement cell with no fluidic blockage. Utilizing SPHERO™ Alignment Particles, the coefficients of variation (CVs), peak channels, and histogram distributions may be measured to find out the alignment and performance of the movement cytometer. Additiona info may be discovered at STN-8: Calibration and Efficiency Monitoring of Movement Cytometer Utilizing SPHERO Calibration Particles.
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Description
Extremely Rainbow Fluorescent Particles (by Spherotech™) are internally labeled with a number of fluorophores. They exhibit full spectrum fluorescence, together with enhanced UV and Far Pink fluorescence. The combination of fluorophores allows Extremely Rainbow particles to be excited at any wavelength from 365 to 650nm. These particles can be found in two diameters, ~3.8µm and ~10.2µm.
Traits
Imply Diameter: ~3.8µm (3.61-3.99µm) or ~10.2µm (8.1 – 12.0µm); see your Certificates of Evaluation for precise measurement.
Focus: ~2 x 106particles/mL
Storage Buffer: Deionized water with 0.02% sodium azide and 0.01% NP-4O
Process
A dilution of 1-2 drops to 1mL will present an enough variety of particles for movement cytometry use.
Notes: Shake vigorously or vortex briefly earlier than use. Inclusion of a small quantity of detergent within the diluent will assist improve the variety of singlets. Diluted particles may be saved within the fridge for future use.
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