BD CaliBRITE beads are designed for use with FACSComp or AutoCOMP software and the FACS family of flow cytometers (FACSCalibur, FACSort, FACScan. values for BD Calibrite beads. To edit, see page A Target file is also created for. HLA-B Although used by. BD FACSComp software, the file is not editable. Product Name: BD CALIBRITE BEADS. Synonyms: BD CALIBRITE BEADS; CALIBRITE BEADS. CAS: MF: MW: 0. EINECS: Mol File: Mol File. BD CALIBRITE .
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The channel separation and PMT voltages for each of the four parameters should be maintained in a daily log to track instrument performance. Reagents are sufficient to perform 25 calibriite. Optimize settings for your sample, as needed. A minimum channel separation must be met for the scatter and fluorescence parameters.
BD CALIBRITE BEADS
Following Calbrite and compensation adjustment, the software calibrtie a Sensitivity Test using the appropriate mixed-bead suspension. For further assistance, contact your BD Biosciences service representative.
Beads used beyond their stability begin to show a decrease in separation between unlabeled and labeled populations, possibly resulting in Sensitivity Test failure.
Concentration values are listed in the following table: This allows cells to be distinguished from sample debris or background signal and for dimly stained cells to be distinguished from unstained cells. Figure 1 through Figure 4 show examples of optimization for two- three- and four-color applications. Always refer to the appropriate application note or reagent IFU.
Use the same staining method and run in parallel with the test samples. Generate a printout of the Sensitivity Test results and keep the printouts in a log book. This instructions for use IFU provides information for two- three- and caibrite setup.
Notice populations with a lower FSC signal than lymphocytes debris, for example can be excluded by increasing the FSC threshold level. One bottle is sufficient to perform 25 tests. Record PMT voltages and channel separations obtained for each parameter in a daily log sheet.
A thorough investigation of sucrose Do not dilute PerCP-Cy5. For information on use, refer to the appropriate instrument manual. UV Bead lab with graph. Mix bead vials by gentle inversion or very gentle vortexing prior to use.
How to make the beaded number line. See Optimization and Quality Control on page 4. Make sure to obtain a full drop of beads.
BD Calibrite PerCP-Cy5.5 Beads
Optimization and Quality Control Because leucocytes have different optical properties than BD Calibrite beads, optimization of instrument settings with cell samples is important.
All forms are provided in stabilized, buffered saline with 0. BD Calibrite beads are used to determine the appropriate compensation settings. The suspensions are stable for a longer period of time in Bead Dilution Buffer. Optimizing Scatter Figure 1 shows a lysed whole blood LWB sample from a normal donor before and after optimization.
NOTE Over a period of time, the fluorescence separation might decrease. FL1, FL2, and FL3 fluorescence sensitivity is determined by measuring the mean channel separation between the signal of cakibrite labeled beads and the unlabeled beads.
BD Calibrite™ Beads
Optimize instrument settings following two-color setup using a blood sample stained with any combination of monoclonal antibodies that identifies separate non-overlapping cell populations, such as FITClabeled and PE-labeled monoclonal antibodies. After the instrument settings have been determined, BD Calibrite beads are used to evaluate instrument sensitivity.
Compensation adjustments for FL1, FL2, and FL3 correct for spectral overlap by shifting the labeled bead populations so they are aligned with the corresponding unlabeled bead populations. If this occurs, manually adjust the settings. Preparation of Test Suspensions Prepare all bead suspensions immediately prior to use. Optimization following three- and four-color setup can vary depending on the application.
It might be necessary to adjust the FSC and SSC amplifiers so that all leucocyte populations are on scale, and to adjust compensation and threshold settings see Figure 1. Adjust fluorescence compensation using Tube B. This product is licensed for sale only for in vitro diagnostics.