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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Beads used beyond their stability begin to show a decrease in separation between unlabeled and labeled populations, possibly resulting in Sensitivity Test failure. Figure 1 through Figure 4 show examples of optimization for two- three- and four-color applications.
BD Calibrite™ Beads
Generate a printout of the Sensitivity Test results and keep the printouts in a log book. Make sure to obtain a full drop of beads. The 3-color kit contains these beads plus a PerCPlabeled bead.
Optimization and Quality Control Because leucocytes have different callbrite properties than BD Calibrite beads, optimization of instrument settings with cell samples is important.
Reagents are sufficient to perform 25 tests. This product is licensed for sale only for in vitro diagnostics. The FSC threshold is adjusted to a level that minimizes background signal if any. NOTE Over a period of time, the fluorescence separation might decrease. Wellington, Auckland, New Zealand bdbiosciences.
BD Calibrite™ – BD Calibrite PerCP-Cy Beads – BD Biosciences
A minimum channel separation must be met for the scatter and fluorescence parameters. If deterioration is suspected, prepare a new bead suspension and check instrument conditions. One bottle is sufficient to perform 25 tests. NOTE Invert bead vials completely when adding a drop to the tube. See Figure 1 through Figure 5 for examples. Adjustment is similar for PerCP-Cy5.
Do not use after the expiration date shown on the label. Beads used beyond their stability begin to show a decrease in separation between unlabeled and labeled populations, resulting in Sensitivity Test failure.
NOTE Different immunophenotyping preparation methods might require different optimization procedures.
BD Calibrite PerCP-Cy5.5 Beads
See examples in Optimization and Quality Control on page 4. Refer to the information appropriate to the instrument setup you are performing. Preparation of Test Suspensions Prepare all bead suspensions immediately prior to use. Gently mix the BD Calibrite bead vials, then add 1 drop of beads to each tube as indicated in the table below.
Always refer to the appropriate application note or reagent IFU. Next, the software adjusts fluorescence compensation using a mixed-bead suspension containing equal amounts of the appropriate BD Calibrite beads. Notice populations with a lower FSC signal than lymphocytes debris, for example can be excluded by increasing the FSC threshold level. 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.
The 2color kit contains three different types of BD Calibrite beads: The drop should be cloudy, indicating the beads are properly mixed. Optimizing Scatter Figure 1 shows a lysed whole blood LWB sample from a normal donor before and after optimization.
Do not dilute PerCP-Cy5. The channel separation and PMT voltages for each of the four parameters should be maintained in a daily log to track instrument performance. Because BD Calibrite beads simulate unstained cells and cells that have been stained labeled with fluorochrome-conjugated antibodies, the beads are calubrite to adjust the instrument settings before cell samples are run on the flow cytometer.
BD CALIBRITE BEADS
The flow cytometer has separate detectors or photomultiplier baeds PMTs that detect light signals. Mix bead vials by gentle inversion or very gentle vortexing prior to use. Optimization following three- and four-color setup can vary depending on the application. Forward scatter FSC and side scatter SSC instrument sensitivity are measured by the mean channel separation between the light-scatter signal calkbrite the beads and background signal electronic and optical.
The following list illustrates PMT light signal detection: After the instrument settings have been determined, BD Calibrite beads are used to evaluate instrument sensitivity. If you require any additional license to use this product and do not have one, return this beadx, unopened, to BD Biosciences, Qume Drive, San Jose, CAand any money paid for the material will be refunded. For further assistance, contact your BD Biosciences service representative.
The decrease in separation for a wide variety of bead lots has been within 2. Label two 12 x mm polystyrene tubes Tube A and Tube B.
Corrective action might be required if the average separation varies by more than 2. If this occurs, manually adjust the settings.