Equipment & Materials; Preparation - Chrono-Log 490 4+4 Instruction Manual

Type:
For testing with 500 µL PRP samples: Five (5) 4.5 mL blue top
evacuated tubes per patient. For testing with 250 µL PRP Samples:
Three (3) 4.5 mL blue top evacuated tubes per patient.
Specimen should be drawn with a minimum of trauma or stasis at
the venipuncture site and anti-coagulated with 3.2% or 3.8%
sodium citrate, in the ratio of one (1) part anticoagulant to nine
(9) parts of blood.
An EDTA blood specimen must be collected from the patient for
hematocrit and platelet count. The blood specimen must be
collected using plastic equipment throughout.
Plastic or non-contact surfaced (siliconized) materials should be
used throughout in order to minimize activation of the platelets
during sample preparation.
Handling Conditions:
Testing can start 30 minutes after venipuncture and continue for
about 2.5 hours after. Specimen should be kept at room
temperature (24° to 27°C).
EQUIPMENT AND MATERIALS:
Equipment:
1. CHRONO-LOG® 490 4+ or 490 4+4 Aggregation system with
internal AGGGRO/LINK® Interface.
2. Windows®-Compatible Computer and AGGRO/LINK® Opti8
software installed.
Materials:
1. P/N 312 Cuvettes
2. P/N 311 Disposable Siliconized Stir bars
3. P/N 322 Calibration Kit containing 1 Black Cuvette and 2
Sealed Water Cuvettes.
4. P/N 331 .5-10 µL Adjustable Pipette
5. P/N 335 Tips for P/N 331
6. P/N 332 10-100 µL Adjustable Pipette
7. P/N 337 Tips for P/N 332
8. P/N 333 100-1000 µL Adjustable Pipette
9. P/N 339 Tips for P/N 333
®
10. P/N 384 CHRONO-PAR ADP
®
11. P/N 385 CHRONO-PAR Collagen
12. P/N 390 CHRONO-PAR ® Arachidonic Acid
®
13. P/N 393 CHRONO-PAR Epinephrine
®
14. P/N 396 CHRONO-PAR Ristocetin
15. P/N 397 Saline (0.9%) 15mL
Avoid blood bank saline because it may be an incorrect
osmolality. Cell counter diluents are not suitable because they
contain EDTA, which inhibits platelet aggregation. Some
infusion salines are inappropriate because they contain benzyl
alcohol (or other preservatives). Such preservatives/additives
inhibit platelet function.
16. P/N 398 Purified Water 15mL.
Should be pyrogen free (ATP free) for reconstituting reagents.
Avoid any sterile water for injection that contains benzyl alcohol
or other preservatives/additives because they inhibit platelet
function.
17. 15 mL conical test tube and cap, or similar (per test subject):
for storing the PRP and PPP specimens.
Document # 49044IM1
Revision 7.5
Dated February 16, 2017
18. Ice bucket: for maintenance of the reagents during the
course of the working day
19. Vortex-type mixer: for mixing the Arachidonic Acid
20. Long-stemmed plastic transfer pipettes to take off PRP and
PPP for placing into 15 mL test tubes.
21. Lintless wipes, such as KimWipes
Gauze squares are NOT suitable.

PREPARATION:

1. Aggregometer
a. Turn on the unit and let it heat up for 10-15 minutes or
until the heater block stabilizes at 37°C.
b. Place P/N 311 Stir bars in P/N 312 Cuvettes.
c. Put cuvettes containing stir bar in the incubation wells
to warm up.
d. When the Model 490 4+4 test channels are set to
Reference Channel 1, a single sample of PPP can be
used as the reference sample for all tests run with the
same patient's blood, so the amount of PPP required is
only enough for one sample, about 500 µL.
2. Preparation of Sample:
a. Mix sample by gentle inversion; DO NOT SHAKE.
b. To prepare the platelet rich plasma (PRP):
1) Centrifuge sample at approximately 100-170g for
15 minutes.
2) Take off the PRP with a polypropylene transfer
TM pipette and place into a polypropylene plastic
tube and add cap.
3) Recap the blue top tubes.
NOTE: Per CLSI Document H58-A, the pH of the PRP sample can
be preserved as follows:
• Place PRP in a plastic tube with limited surface area-to-
volume ratio (place large volume of PRP in a small tube)
Cap the PRP tube as PRP in uncapped tube undergoes a rise
•
in pH due to diffusion of CO
• Avoid frequent mixing/agitation of PRP
Introduce PRP directly into the tube and don't allow it to
•
flow down the sides of the tube.
4) Properly label the tube, include the patient's
name and sample type. Parafilm or cap the top.
Keep at room temperature (24°C to 27°C).
c. To prepare the platelet poor plasma (PPP):
1) Place blue top tubes into centrifuge.
2) Centrifuge sample at approximately 1500-2400 g
for 20 minutes.
3) Take off the PRP with a polypropylene transfer
pipette and put it into a polypropylene plastic
tube.
4) Properly label the tube, include the patient's
name and sample type. Parafilm or cap the top.
Keep at room temperature (24°C to 27°C).
When ready to begin testing, dispense one aliquot per channel of
adjusted or non-adjusted PRP of 500 µL volume into P/N 312
cuvettes with stir bars. Warm for a minimum of 3 minutes. It is
not recommended to incubate a sample beyond 30 minutes.
3. Reagent Preparation
The following reagents are sourced from:
Chrono-log Corp.
2 W. Park Road
Havertown, PA 19083 USA
Tel: 610-853-1130
8
®
.
19
from plasma
2