Chrono-Log 490 4+4 Instruction Manual page 13

NOTE: One SELECT and SET pushbutton controls two channels in
tandem. For example, Channels 1 & 2, Channels 3 & 4, Channels
5 & 6 and Channels 7 & 8 are set in pairs.
4. Place the cuvettes containing 500 µL PPP in the reference
well(s) ...stir bar is not required. Check for bubbles and be
sure to wipe cuvette with a clean KimWipe
5. Place 500 µL or 250 µL Platelet-RICH-Plasma (PRP) into pre-
warmed cuvettes with stir bars. [Prepare (1) for each test
channel.] Incubate for three (3) minutes in incubation wells
6. Start AGGRO/LINK® Opti8 TM
select TEST PROCEDURE under the AGGREGOMETER
window. Under PROCEDURE NAME, set-up or load the
procedure that corresponds to the reagent and method
being used. Optical tests with PRP should run for a
minimum of 5 minutes. Slope length can be set from 16 to
99 seconds. Chrono-log standard setting is 16 seconds. Click
OK.
7. Select "RUN NEW TEST" under AGGREGOMETER window.
Patient information page will appear. Patient information
can be completed at this time or can be entered during or
after test is completed. [Click on EDIT then TEST
INFORMATION] When "Run Same Test" is selected, patient
data from previous test will be used.
NOTE – Patient data from Trace 1 can be copied to other
tracings. Click on Select All if testing same donor in all channels
or Click on appropriate Trace Nos. and then Click on COPY.
NOTE – The Test Identification field at the top of the page is used
to identify and select test(s) in the Test Directory. For Clinical
testing, place patient identifier in this field.
8. Place the cuvettes containing PRP in the PRP wells (One for
each Optical Channel). Check for bubbles and be sure to
wipe cuvette with a clean.
9. Click OK to begin test. Click on color-coded "Activate" bar for
each channel.
10. Push the SET BASELINE buttons for each channel. The
tracing should move to 100% when the button is depressed
and to 0% when the button is released, using the numbers
on the left side of the graph. Be sure to press and hold the
Baseline button until the tracing reaches 100% of the graph
and then release.
NOTE: The Graph Range time can be adjusted under VIEW and
then Set Graph Range. When the test tracings have reached the
end of the Graph Range, additional time is added automatically
in one minute increments, up to a total of 60 minutes.
11. Monitor tracings for stability.
12. When tracing(s) have reached stability, take up the
appropriate reagent and click on the color-coded "Start" bar
for Channel 1. [Or ... See NOTE 2 below].
13. Add reagent
a. Use 1 µ µ µ µ L of Collagen with 500 µL sample volume or
0.5 µ µ µ µ L with 250 µL sample for 2 µ µ µ µ g/mL final
concentration...or for a 5 µ µ µ µ g/mL final concentration
use 2.5 µ µ µ µ L with 500 µL sample volume or 1.25 µ µ µ µ L with
250 µL sample.
NOTE: As shown above, reagent volumes are cut in half when
testing with 250 µ µ µ µ L PRP samples.
Document # 49044IM1
Revision 7.5
Dated February 16, 2017
®
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for Windows ® program and
b. Use 5 µ µ µ µ L of ADP with 500 µL sample volume for a final
concentration of 10 µ µ µ µ M...or 2.5 µ µ µ µ L for 5 µ µ µ µ M.
Use 5 µ µ µ µ L of Arachidonic Acid with 500 µL sample
c.
volume for a final concentration of 0.5 mM...or 2.5 µ µ µ µ L
for 0.25mM.
Use 5 µ µ µ µ L of Ristocetin with 500 µL sample volume for a
d.
final concentration of 1.25 mg/mL.
e. Use 2 µ µ µ µ L of Ristocetin with 500 µL sample volume for a
final concentration of 0.5 mg/mL (Type 2B vW).
f. Use 2.5 µ µ µ µ L of Epinephrine with 500 µL sample volume
for a final concentration of 5 µ µ µ µ M.
NOTE – CHRONO-PAR® Reagents do not require preparation of
multiple stock solutions. To change final concentration, adjust
pipette volumes as shown above.
14. Repeat steps 12 and 13 for each test channel.
15. Allow all tests to run for a minimum of five (5) minutes.
NOTE: The "Activate" bar in Step 9 provides the ability to control
each channel individually and to stagger the start of each test.
NOTE: To have the baseline setting visible on the final printout,
when a tracing has reached stability, Click on Aggregometer ...
Reset Channel ... and select appropriate Channel #. Click on the
"Activate" bar and reset the baseline. Then, click on the "Start"
bar and add the appropriate reagent for that Channel.
16. While tests are running, prepare one test cuvette for each
channel (Cuvette, stir bar and 500 µL or 250 µL PRP) & begin
three (3) minute incubation.
17. There are a number of Options to Stop Tests as each test can
be stopped individually or all tests stopped at the same
time, as follows:
a. To Stop Tests individually, click on Aggregometer, click
on Stop Test and select the appropriate Channel.
b. STOP Icon at top of screen stops all tests at the same
time.
18. Clicking on the "Start" bar prior to adding the reagent sets
the Start and Stop time for each channel. If there is a
baseline shift or some other artifact that should not be
included in the final calculation, Click on EDIT and SET START
& STOP TIMES (or use ICONS at top of screen).
a. If only calculating Slope and Amplitude:
1) A small box will appear with Trace 1 selected.
2) If the Automatic Start Time Feature was used
before the addition of the reagent, a dotted line
will appear on the screen at that point (Start
Line) with another dotted line (Stop Line) placed
"X" number of minutes after (Test Time set up on
procedure page).
3) If required, Start & Stop times can be moved:
a) Individually – Left Click, Hold and Drag to
new position. (Be sure Start Line is placed
on stable baseline just before or just after
adding the reagent. Stop Line should be
placed five (5) minutes after the Start
Time.)
b) Simultaneously – Place cursor near Start
Line. Right click, Hold & Drag Start Line to
new position. Stop Line will move in
tandem.
4) Click on other Tracings and repeat, if needed.
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