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COULTER® CYTO-TROL™
Control Cells Kit
For In Vitro Diagnostic Use
extremely toxic compound. Azide compounds should
be flushed with running water while being discarded.
These precautions are recommended to avoid
deposits in metal piping in which explosive conditions
can develop. If skin or eye contact occurs, wash
extensively with water.
3. Never pipet by mouth and avoid contact with skin and
mucous membranes.
4. CYTO-TROL Control Cells should not be used as a
calibrator.
5. The reagents are for flow cytometry use only.
6. Results obtained with flow cytometry may be
erroneous if the laser is misaligned or the gates are
improperly set.
7. Do not freeze reagents.
8. Avoid microbial contamination of reagents or
erroneous results may occur.
9. Harmful if swallowed.
10. After contact with skin, wash immediately with plenty
of water.
INTENDED USE
STORAGE CONDITIONS AND STABILITY
The COULTER CYTO-TROL Control Cells Kit is used to
assess the activity of monoclonal antibodies by flow
cytometry.
Do not use reagents beyond the expiration date printed on
the kit label. Unopened reagents are stable to the
expiration date on the kit label when stored at 2-8°C.
Reconstituted CYTO-TROL Control Cells are stable up to 24
hours when stored at 2-8°C. Do not freeze. Bring reagents
to 20-25°C prior to use.
6604248 - 50 tests
PN 4236083-V
SUMMARY AND EXPLANATION
CYTO-TROL Control Cells are a lyophilized preparation of
human lymphocytes that exhibit surface antigens
detectable with monoclonal antibodies. The cells are
isolated from peripheral blood and express antigens
representative of those found on normal lymphocytes.
CYTO-TROL Control Cells are compatible with directly
conjugated (FITC, RD1, ECD, PC5) Coulter monoclonal
antibodies listed in the Table of Expected Assay Values
and Ranges given below.
EXPECTED RESULTS
*The mean assay values provided for each COULTER
monoclonal antibody are derived from triplicate analyses
of CYTO-TROL Control Cells performed on calibrated flow
cytometers with set parameters and specific laboratory
conditions. Your values may vary depending on your
laboratory’s operating conditions. The absolute measure
of your performance should be based on Coulter’s
Interlaboratory Quality Assurance Program (IQAP) peer
group analysis of the lot’s mean and CV as compared to
that of an individual instrument. Percent positives were
determined from cursors set to establish 98% (nominal)
nonspecific staining using the Isotypic Control.
**Cursor adjustment, which may be performed to separate
positive and negative populations when analyzing blood
samples with these antibodies, was not performed in this
situation.
STATEMENT OF WARNINGS
WARNING: POTENTIAL BIOHAZARDOUS MATERIAL.
Each human donor unit used in preparation of this material
was tested by an FDA approved method for the presence
of the antibodies to Human Immunodeficency Virus (HIV-1
and HIV-2) and Hepatitis C Virus (HCV) as well as for
hepatitis B virus surface antigen and found to be negative
(were not repeatedly reactive).
1. Because no test method can offer complete assurance
that hepatitis B virus, Human Immunodeficiency Virus
(HIV-1 and HIV-2), or other infectious agents are
absent, this specimen/reagent should be handled at
Biosafety Level 2, as recommended for any potentially
infectious human serum or blood specimen in the
Centers for Disease Control/National Institutes of
Health manual “Biosafety in Microbiological and
Biomedical Laboratories,” 1988.
2. The reagents contain sodium azide. Sodium azide
under acid conditions yields hydrazoic acid, an
Staining Procedure for CYTO-STAT Monoclonal
Antibodies:
1. Add 10 µL of monoclonal antibody to 100 µL of
reconstituted CYTO-TROL Control Cells. Mix gently.
2. Incubate at room temperature for a minimum of 15
minutes.
3. After incubation, add 1 mL of phosphate buffered
saline (pH=7.2), PN 6603369, to each tube. Mix gently.
4. Analyze on a flow cytometer.
RECOMMENDED FLOW CYTOMETRY
COLLECTION PROCEDURE FOR
LYMPHOCYTE MARKERS
1. Ensure the flow cytometer is properly aligned and
standardized for light scatter and fluorescence
intensities according to manufacturer’s
recommendations (refer to Instrument Product
Manuals).
2. Collect side scatter [SS] or log side scatter [LSS] vs.
forward side scatter [FS] histogram.
3. Set a bitmap by gating on the major population of cells
observed (see Figure 1).
Figure 1. A two-parameter LSS vs. FS histogram
to identify lymphocytes.
EVIDENCE OF DETERIORATION
If CYTO-TROL Control Cells are not within the expected
ranges given on the Table of Expected Assay Values and
Ranges or show a loss or shift of light scatter:
1. Review the operating procedure of the instrument.
2. Review operator technique.
3. Assay an unopened vial of CYTO-TROL Control Cells
following the package insert.
4. Check for product deterioration which may appear as
clouding or clumping of the reagents.
REAGENT PREPARATION
Do not reconstitute CYTO-TROL Control Cells until ready to
use.
1. Reconstitute by pipetting 1 mL of Reconstitution Buffer
into a vial of cells.
2. Mix gently by inversion.
3. Allow cells to stabilize for 10 minutes before use.
PROCEDURE FOR
IMMUNOFLUORESCENCE CELL SURFACE
STAINING OF CYTO-TROL CONTROL CELLS
WITH COULTER MONOCLONAL ANTIBODY
REAGENTS
The recommended volume is 100 µL of reconstituted
CYTO-TROL Control Cells per test.
The use of appropriate Isotypic Controls is
recommended to monitor levels of nonspecific
staining.
NOTE: CYTO-TROL Control Cells are not tested for use in
whole blood lysing procedures. Use of COULTER
Q-PREP™/IMMUNOPREP™, MULTI-Q-PREP™/IMMUNOPREP
or TQ-PREP™/IMMUNOPREP Reagent Systems for red
blood cell lysing may produce variations from assigned
values.
Staining Procedure for Non-CYTO-STAT
Monoclonal Antibodies:
1. Substitute 100 µL of reconstituted CYTO-TROL Control
Cells for the sample of interest and complete the
procedure for immunofluorescence cell surface
staining with the applicable non-CYTO-STAT
monoclonal antibodies.
2. Analyze on a flow cytometer.
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4. Collect log fluorescence for this population of cells.
Percent positive cells can be determined by integrating
a clear fluorescence peak (single color) or by setting
quadrant statistics (dual color). Refer to the flow
cytometer reference manual for detailed instructions.
With certain low-density markers, some laboratorians may
elect to move the cursors set with Isotypic Controls. For
best results, analyze on the single-parameter histograms
and set the cursors to the appropriate demarcation
between the visually positive and negative regions.
PRODUCT AVAILABILITY
COULTER CYTO-TROL Control Cells Kit
PN 6604248 - 50 tests
REAGENTS CONTAINED IN KIT
CYTO-TROL Control Cells
5 vials (≥3.5 X 106 cells/vial)
Reconstitution Buffer
5 x 1 mL/vial
Patent No. 5,059,518
For additional information within the US, call
1-800-526-7694. Outside the US, contact your local
Coulter Representative.
TRADEMARKS
COULTER, COULTER CLONE, CYTO-STAT, CYTO-TROL,
IMMUNOPREP, MULTI-Q-PREP, Q-PREP, and TQ-PREP are
trademarks of Beckman Coulter, Inc.
Beckman Coulter, Inc.
4300 N. Harbor Blvd.
Fullerton, CA 92835
www.beckmancoulter.com
Beckman Coulter Ireland Inc.
Mervue Business Park,
Mervue, Galway,
Ireland (353 91 774068)
Printed in USA
Made in USA
© 2002 Beckman Coulter, Inc.
All Rights Reserved.
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