The Jackson Laboratory

Prepared by:

PM

Date Prepared:

March 20, 2001

Code:

MP-006

Reviewed by:

Revision #:

4.1

Date Revised:

February 2, 2006

0.0

Abstract: The ADVIA 120 Hematology System is a diagnostic instrument that analyzes whole blood to provide the following types of results:

• Complete Blood Count (CBC) including White Cell Differential
• Reticulocytes (Absolute count, percent and indices)

1.0

Instrumentation layout: n/a

2.0

Reagents and expendables:

• ADVIA TESTpoint Hematology Controls
• ADVIA 120 autoRETIC Reagent T01-3622-01
• Perox Sheath T01-3633-01
• Perox 1 T01-3630-01
• Perox 2 T01-3631-01
• Perox 3 T01-3632-01
• RBC/PLT T01-3627-01
• HGB T01-3628-01
• BASO T01-3629-01
• AutoRETIC T01-3622-01
• EZ KLEEN T01-3624-01
• Defoamer T01-3625-04
• Sheath/Rinse T01-3664-01

All reagents are ordered from:
Bayer
Diagnostics Division
Bayer Corporation
511 Benedict Avenue
Tarrytown, NY 10591-5097
Phone: (914)631-8000
Fax: (914)524-2132
http:\\www.bayerdiag.com

Maintenance and upkeep provided by the Diagnostic Laboratory at The Jackson Laboratory.

3.0

Setup: You need to contact the Diagnostic Laboratory to be sure that ADVIA is working and that controls have been run. Once ADVIA is operating properly, go and obtain the blood samples.

4.0

Run: Prepare Eppendorf tubes with 20 µl of 20% EDTA. Use microhematocrit tubes coated with EDTA. Obtain and place 250 µL blood in Eppendorf tubes. Use a wooden toothpick to check for clots. If any clots are present discard the sample.

5.0

Clean up: Dispose of used Eppendorf tubes by placing them in a burn bin or hazardous waste container. Wipe up any blood spills with 70% ethanol.

6.0

Data reduction: To save your specific file be sure to: Log On - Click New File - Save. At the end of the run, you can retrieve your data from ADVIA with a floppy disk by doing the following:

• Press and Hold Control then press Escape.
• Click on the square in the left lower corner to open to Explorer.
• Click on Explorer - go to Bin - to Export Data - Look for the date of your Data File.
• Highlight file and drag item to floppy disk item on left side. If done correctly, you will see "files" flying from one folder to another.

When completed you will see original Explorer screen. Go to Close, highlight task bar for ADVIA and this will bring you back to Manual Sample ID screen. Highlight Sample Control Panel - Rev/Edit - click OK in "Unmatched Tubes" box - Look through all data by clicking the down arrow button - click OK - complete this procedure for all samples. Then Log Off and click Standby (left lower corner).

7.0

Safety: Wear gloves when dealing with blood samples or controls. ADVIA TESTpoint Controls contain human blood cells in a preservative medium.

8.0

Time and capacity: Calibration of controls takes about 25 minutes. Aside from the time it takes you to eyebleed, running samples takes about 1 minute per sample when done through open tube aspiration wand.

9.0

Protocols:
Click Standby Button (located in left lower corner). Put control in incubator to warm.
Start: Insert name and ID #; click Log On.
Go To: Customize - system setup - systems options
Go To: Run screen options - print none - save - routine operations
Go to: Data options (left side of upper screen)
Find: Raw data directory name (it should say C:\RAW_DATA/HLB).
Click Save. You should see: System Options - Data 0 System Options Saved
Click OK
Go To: Routine Operations (left upper corner).
Click Manual Sample ID. On this screen, click in the following information:

Before attempting to place sample, make sure top left corner says Ready to Run. Get controls and be sure to roll vial between hands to mix sample well.
Two Ways to Place Sample in Advia:

1.      Without removing cap of vial - After vial has been warmed and rotated in hands to mix contents, place vial into open port access located next to wand device. Push vial (rubber stopper down) into open port access. Observe green light flashing and then stop. You will feel the vial pop up when sufficient sample has been collected.

2.      Removing cap of vial - Unscrew cap and slide wand into sample. Be sure the wand is set into the sample. Push lever behind wand, and sample will be aspirated into Advia.

If there was sufficient sample, Advia will say Running in top left hand corner. If there wasn't sufficient sample, or if there are clots in the sample or an air gap as the sample was aspirated, Advia will say Aspiration Failed. You will not get any results.

All controls are being run by the Diagnostic Laboratory Staff.  Please contact them at ext. 1270 for questions and formal training on the use of the ADVIA.

The ADVIA must have the PRIMER run before you run your samples.

1st: The primer being used is called Sheath Reagent and is located on the right side of the ADVIA.  Click on the following settings for Primer rinse:

Go To: Manual Sample ID and click Sample Type to Control. Activate bar code on each tube with hand Opticon device.
2nd: Controls:

After each sample has run, you will see a lab screen pop up. Click Hide to go back to Manual Sample ID Screen. Then click QC at bottom of screen and review the lab results. Note: If you see Yellow, it could be that the sample was not mixed or warmed up enough. Re-do the sample. If you see Red, there is a problem with the sample and Sue Grindle (x6203) needs to be contacted.
Go To: Routine Operations - Manual Sample ID - Change Sample Type to Primer
3rd: Primer Again (to wet system). Change sample type to Control.
4th: Controls:

Again click on Hide and return to Manual Sample ID Screen. Check lab values on QC screen. Observe for yellow or red flags. If all flags for controls are normal, proceed with eyebleed samples.
For Eyebleed Samples: Go to Manual Sample ID, click New File - Save - OK.
Make sure printer is set to print lab sheets by going to Customize - System set Up - System Options - Run Screen Options. Click Print All; click Save-OK; then go to Routine Operations and click Manual Sample ID.

The primer being used is called Sheath Reagent and is located on the right side of the ADVIA.  Click on the following settings for Primer rinse:

Put in sample id by clicking Field and then click OK or press Enter. Click in the following information:

Advia should say Ready to Run. Type strain name like ####F/M##.

10.0

Notes: n/a