SPHEROT -Ultra Rainbow Fluorescent Particles
New motion cytometers, with fluorescent channels from the UV to Far Pink, and corresponding fluorescent conjugates
in the mean time are accessible. In consequence, we now have developed the Extraordinarily Rainbow Fluorescent Particles with enhanced UV
and Far Pink fluorescence depth. The Extraordinarily Rainbow Fluorescent Particles embody a single peak and are
designed for checking the optical alignment of any motion cytometer in all channels.
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%) throughout 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 subsequent:
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 indicate channel number of the peak is in a predetermined place on each histogram of curiosity. The histograms on the sooner net 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 widespread or preset values, which are determined by not lower than one months worth of data, additional calibration or alignment procedures have to be carried out in response to the instrument operation handbook.
Motion Cytometry Alignment Particles
Spherotech offers all types of particles for determining if a motion cytometer is accurately 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 search out out the alignment and efficiency of the motion cytometer. Additiona data could also be found at STN-8: Calibration and Effectivity Monitoring of Motion Cytometer Using SPHERO Calibration Particles.
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Description
Extraordinarily Rainbow Fluorescent Particles (by Spherotech™) are internally labeled with quite a few 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
Indicate 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 throughout the diluent will help enhance the number of singlets. Diluted particles could also be saved throughout the fridge for future use.
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