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Short Instruction Manual
multiplate
Short Instruction Manual
CONTENTS
Getting started Reagent handling Reagent storage conditions Handling of pre-warmed diluent tubes Performing tests using the electronic pipette Electronic pipette: avoiding handling errors Avoiding operator errors Performing tests using a manual pipette Test protocols for hirudinized / heparinized blood Test protocols for citrated blood Multiplate® – Parameters Pipette maintenance
4 5 6 7 8
10 11 12 14 15 16 17
Please note that the Multiplate® platform and reagents are not FDA cleared for in vitro diagnostic use and therefore for research use only.
4
The Multiplate® Software uses the Windows XP operating system. A password is required in order to start (Windows-Login):
username: multiplate password: multiplate
The software then launches automatically and begins heating the Multiplate® analysis area to 37°C. The actual temperature may be seen on the status bar. The instrument will reach the specified temperature range within 10 minutes.
The manufacturer recommends that an electronic control is performed before the first measurement of each working shift (drop down menu: Multiplate > Electronic control). The system must run for at least 20 minutes before starting an electronic control.
Prepare the reagents as described on page 5. Fill the pre-warming tubes with sample diluent (0.9 % saline solution or NaCl / CaCl2) place into position in preheating area of the Multiplate® and open the tips tray.
Before using the electronic pipette inspect the filter and replace it if necessary (details page 18).
Prepare your test cells (check expiry date), and place the blood sample at room temperature (e.g. in the reagent holder).
Multiplate® – Getting started
Standard precautions should always be followed when dealing with patient samples and reagents. The disposal of materials must be performed in accordance with local regulations.
The Multiplate®-System user interface
menu barstatus barcommand bar
Channel display for 5 channels
5
Multiplate® – Reagent handlingUnopened reagent vials
Store lyophilized reagents at 2-8°C. They are stable until the expiry date stamped on the label.
Reagent preparation
Reconstitute with 1.0 ml of purified water and swirl gently to mix.
Aliquot into single day portions
If the reagent is used over several days it is advisable to aliquot into individual tubes (e.g. 5 x 200 μl or 10 x 100 μl). Aliquot tubes must be closed between measurements to avoid evaporation as this may create concentration fluctua-tions. For this reason the manufacturers do not recommend aliquots of less than 100 μl.
Once the reagent is dispensed into the aliquot tubes replace the safety cap tightly. The following page describes storage conditions.
The vials should be marked with the date of reconstitution. Do not refreeze reagents. Original vials of reconstituted reagent should be refrigerated unless in use, and stored as per manufacturers instructions.
Original vial
Aliquot tube
Security seal
6
Once reconstituted the reagents are stable for 7 days at 2-8°C (in the original vial). Storage in aliquots at < -20°C increases stability to 4 weeks.
Multiplate® – Reagent storage conditions
ADPtest RISTOtest TRAPtest ASA Control Prostaglandin E1 (PG)
Once reconstituted the reagent is stable for 24 hours at 2-8°C.
Storage in aliquots at < -20°C increases stability to 4 weeks.
ASPItest
Once reconstituted the reagent is stable for 7 days at 2-8°C (in the original vial).
Do not freeze the COLtest reagent!
COLtest
For lot-specific information always read the current package insert.
7
Multiplate® – Handling of pre-warmed diluent tubes Each Multiplate® unit is supplied with 10 empty preheating tubes with screw caps.
Fill the tube with sample diluent (0.9 % saline solution or NaCl / CaCl2) at the beginning of a Multiplate session.
Contamination of sample diluent or unapproved preservatives can influence aggregation results: therefore tubes should be changed regularly or when suspect of contamination. Only use sterile 0.9 % saline solution or manufacturer’s NaCl / CaCl2 diluent.
Pre-warming diluent tubes (REF MP0090) must be used, otherwise the correct temperature of sample diluent can not be guaranteed. This may lead to erroneous test results.
The tubes are also directly available from Sarstedt AG & Co: Part no. 60.550.100 (www.sarstedt.com).
8
Select the button F1: Auto Pipette
Multiplate® - Performing tests using the electronic pipette 1 / 2
Instructions are displayed on the screen. Press the blue start button on the pipette to execute each pipetting step.
The active step is highlighted with a green arrow. Completed steps have a checkmark, steps to be performed are marked with blue dots.
Place test cells into the targeted channels.
Connect the sensor cable to the test cell. Confirm with the prompt Next.
Enter the patient ID and test name.
Up to 5 tests may be started simultaneously. Confirm with the prompt start auto-pipette.
9
Multiplate® – Performing tests using the electronic pipette 2 / 2
After addition of the diluent solution and the blood sample to the targeted channels a 3 minute incubation begins automatically.
An audible alarm indicates the end of the incubation time and that the activator must be added to each channel. After 6 minutes / end of test:
▪ Print the results if required (F6: Print), select the appropriate channels and confirm. After the results are printed the following icon appears in the channel window:
▪ Dispose of the test cells appropriately.
▪ Clear channels (F7: Clear channels), select the appropriate channels and confirm. This icon indicates that the channel is available:
The checkmark icon in the channel window indicates the end of the test.
Addition of the activator triggers an automatic start of the channel.
In the channel window the icon displayed to the right indicates that the test is running.
10
Add the activator (e.g. ASPItest reagent) to the bottom of the test cell.
Execute all of the pipetting steps sequentially, according to the instructions on the screen.
Continuously pressing the Start key (blue) during communication between the pipette and the instrument may interfere with proper communication: therefore it is necessary to wait until the next instruction appears on the screen before proceeding.
The previous step must be completed and the next pipette instruction displayed on the screen before using the eject function. Care and patience are required when handling the electronic pipette.
Visual control of pipette volumes:
A visual inspection of the aspirated volume in the pipette tip is recommended before adding the solution to the test cell.
This may avoid an operating error due to aspirated air or another potential malfunction.
The images illustrate the accurate pipette tip fill levels for volumes of 20 μl and 300 μl.
Multiplate® – Electronic pipette: avoiding handling errors
20 µl 300 µl
11
Multiplate® – Avoiding operator errors
Store blood samples at room temperature, do not heat the sample.
Sample diluent (0.9 % saline solution or NaCl / CaCl2) must be pre-heated for a minimum of 10 minutes before use.
After manually pipetting the activator it is important to trigger the start of the test immediately.
Pipette the activator into the bottom of the test cell.
Don‘t remove the stir bar! Ensure that it is present in the test cell.
Place the test cell firmly into the measuring position.
WRONG! RIGHT!
12
Multiplate® – Performing tests using a manual pipette 1 / 2
Start the incubation timer (warming and equilibration): Select the button F2: Start Timer. A timer window appears on the screen and a 3 minute countdown will begin.
Pipette 300 µl saline solution 0.9 % + 300 µl hirudinized / heparinized blood or 300 µl NaCl / CaCl2 + 300 µl of citrated blood into the test cell (for details see page 14/15)
Place the test cell(s) into the measuring position(s). Attach the sensor cable(s), ensure that each cable plug is connected.
13
After 6 minutes / end of test:
▪ Print the results if required (F6: Print), select the appropriate channels and confirm. After the results are printed the following icon appears in the channel window:
▪ Dispose of the test cells appropriately.▪ Clear channels (F7: Clear channels),
select the appropriate channels and confirm. The icon to the right indicates that the channel is available:
The icon in the channel window will change to indicate the end of the test.
This icon appears when the channel is actively running a test:
Enter a patient ID (F4: Enter ID) and select a test name (T: Test selection).
Steps 4 and 5 must be performed for each channel seperately:
Multiplate® – Performing tests using a manual pipette 2 / 2
After incubation: Pipette the activator into the bottom of the test cell and start the test by selecting the F3: Start Test or <F3> button.
Select a channel (using the mouse or the keys 1-5) and confirm with Start or Enter.
Analysis begins for the selected channel.
14
Multiplate® – Test protocols for hirudinized / heparinized blood
20 µl TRAPtest
20 µl ADPtest
20 µl ADPtest
For ADPtest HS
20 µl ASPItest
20 µl COLtest
12 µl RISTOlow
++
20 µl Prostaglandin E1
+
50 µl RISTOhigh
300 µl saline solution 0.9% (37°C) + 300 µl hirudinized / heparinized blood
3 minute incubation
Measuring time: 6 minutes
Measuring time: 6 minutes
3 minute incubation
(HS = high sensitivity)
15
Multiplate® – Test protocols for citrated blood
20 µl TRAPtest
20 µl ADPtest
+
20 µl COLtest
Measuring time: 6 minutes
3 minute incubation
300 µl saline solution 0.9% / 3 mM CaCl2 (37°C) + 300 µl citrated blood
300 µl saline solution 0.9% (37°C) + 300 µl citrated blood
20 µl ASPItest
12 µl RISTOlow
50 µl RISTOhigh
+
Measuring time: 6 minutes
3 minute incubation
16
The Multiplate® continuously records the platelet aggrega-tion. The attachment of platelets onto the Multiplate sensors generates a change in impedance which is transformed into arbitrary aggregation units (AU) and displayed as a curve.
The most important parameter is the area under the aggre-gation curve (AUC). It is affected by the total height of the aggregation curve as well as by its slope and is best suited to express the overall platelet activity. Two more parameters are calculated for research use: the aggregation reflects the height of the curve and the velocity reflects the maximum slope of the curve.
As represented in the diagram to the left, two curves are assessed using the two independent sensors in the Multiplate® test cell. The parameters calculated by the software represent the mean val-ues of the combined data from the two curves.
To the left of the curve a black bar graphically indicates the AUC (see screen depiction below right). The dark green portion represents the reference range as defined in the software for each test type by the user.1
Using the key <F9> or the button F9: curve mode it is possible to toggle between the curve and the area display.
Multiplate® – Parameters
Twin sensors for duplicate measurements
Agg
rega
tion
(AU
)
Time (min)
Velocit
y (Vel)
Area under the curve AUC plot
1 The display of the reference ranges is not activated in the default software settings.
17
Multiplate® – Pipette maintenance
The electronic pipette filter should be inspected frequently. Cleanliness helps to prevent clots forming on the filter and maintain accurate fill levels.
Only use REF MP0033 / MP0034 filters as supplied by the manufac-turer.
When a filter change is required forceps are provided to remove the contaminated filter.
Special care should be taken when pulling the filter from the pipette cone.
Use forceps to handle the new filter.
Insert it gently but firmly into the end of the pipette cone.
Clean filters will extend the lifetime of the electronic pipette.
V4.0-RUO REF MP0921-US
Verum Diagnostica GmbHReichenbachstrasse 2780469 Munich Germany
MANUFACTURER
Phone: +49 - 89 - 125556 - 0Fax: +49 - 89 - 125556 - 11E-mail: [email protected]: www.multiplate.us