Ultra Rainbow Fluorescent Particles SPHERO
Motion Cytometry Alignment Particles
Spherotech offers every kind of particles for determining if a motion cytometer is appropriately aligned and has a transparent motion cell with no fluidic blockage. Using SPHERO™ Alignment Particles, the coefficients of variation (CVs), peak channels, and histogram distributions could also be measured to seek out out the alignment and efficiency of the motion cytometer. Additiona information could also be found at STN-8: Calibration and Effectivity Monitoring of Motion Cytometer Using SPHERO Calibration Particles.
Spherotech Cat. No. URFP-30-2 Instructions for use:
A. Preparation of Particles
1. Vortex the particles vigorously
2. Add 2 to 4 drops of particles to 1mL of sheath fluid or DI water. The inclusion of a small amount of detergent (~0.01%) inside the dilution buffer will enhance the proportion of the singlet inhabitants.
B. Every day Alignment
To search out out the optical alignment of the system perform the following:
1. Set a keep gate for the singlet inhabitants on the FSC vs SSC histogram to exclude aggregates
2. Modify the Purchase and Extreme voltage so that the suggest channel number of the peak is in a predetermined place on each histogram of curiosity. The histograms on the sooner internet web page will be utilized as a info.
3. Accumulate 5000 events contained within the gate
4. Report the % CV and Extreme Voltage for all fluorescence channels of curiosity.
5. Use a computer program resembling Excel to generate the Levy Jennings graphs.
NOTE: If the values on any parameter exceed these of the day-to-day frequent or preset values, which can be determined by not lower than one months worth of knowledge, additional calibration or alignment procedures have to be carried out in response to the instrument operation handbook.
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Description
Extraordinarily Rainbow Fluorescent Particles (by Spherotech™) are internally labeled with quite a lot of fluorophores. They exhibit full spectrum fluorescence, along with enhanced UV and Far Pink fluorescence. The mixture of fluorophores permits Extraordinarily Rainbow particles to be excited at any wavelength from 365 to 650nm. These particles will be present in two diameters, ~3.8µm and ~10.2µm.
Traits
Suggest Diameter: ~3.8µm (3.61-3.99µm) or ~10.2µm (8.1 – 12.0µm); see your Certificates of Analysis for exact measurement.
Focus: ~2 x 106particles/mL
Storage Buffer: Deionized water with 0.02% sodium azide and 0.01% NP-4O
Course of
A dilution of 1-2 drops to 1mL will current an sufficient number of particles for motion cytometry use.
Notes: Shake vigorously or vortex briefly sooner than use. Inclusion of a small amount of detergent inside the diluent will help enhance the number of singlets. Diluted particles could also be saved inside the fridge for future use.
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