Hematology Procedure Manual
Principle The Coulter AcT2 Hematology Analyzer performs a Complete Blood Count (CBC), Platelet Count, and a Three-Part Differential. Whole blood is aspirated, diluted, and then divided into two samples. One sample is used to analyze the red blood cells and platelets while the second sample is used to analyze the white blood cells and hemoglobin. Electrical impedance is used to count the white blood cells, red blood cells, and platelets as they pass through an aperture. As each cell is drawn through the aperture, a change in electrical resistance occurs generating a voltage pulse.
The number of pulses during a cycle corresponds to the number of cells counted. The amplitude of each pulse is directly proportional to the cell volume. Lyse reagent is added to the diluted sample and used to count the white blood cells. After the white blood cells have been counted and sized, the remainder of the lysed dilution is transferred to the Hgb Flow Cell to measure Hemoglobin concentration. The AcT2 uses electronic sizing to determine a three part automated differential from the WBC histogram. The percentage and absolute counts are determined for lymphocytes, neutrophil, and monocytes. The AcT diff2 analyzer has two operating modes: Open Vial Whole Blood and Closed Vial Whole Blood.
Whole blood samples can be analyzed in either mode. Prediluted samples can be analyzed at the Open Vial Station. Specimen Collection and Handling HANDLE BLOOD AS A POTENTIAL BIOHAZARD CAPABLE OF TRANSMITTING INFECTION. ALWAYS WEAR PROTECTIVE GLOVES AND LAB COAT WHEN PROCESSING SPECIMENS. Draw specimen in into a lavender-top vacutainer tube containing K2EDTA.
Hematology Sysmex Procedure Manual
Thoroughly mix blood with EDTA anticoagulant. If hemolysis or small clots are observed, discard specimen. Mix venous blood sample at least 8 times by hand inversion. Gently turn capped sample upside down then back straight up. Alternatively, use a mechanical mixer for at least 5 minutes. Do not test samples that are incorrectly filled or that are clotted. If hemolysis or small clots are observed, discard specimen.
Analyze venous blood samples within 24 hours of collection. Do not refrigerate samples for platelet and differential counts. If platelet or differential results are not required, store anticoagulated whole blood specimens at 2- 8 C. Warm samples to room temperature (16-35 C or 61-95 F) before testing.
EQUIPMENT AND MATERIALS Equipment Performance Parameters The AcT diff2 Analyzer operates at ambient temperature (16- 35 C or 61- 95 F) at humidity no higher than 85% without condensation. Materials, Reagents COULTER diff AcT Tainer Reagent Kit which contains diluent (Reagent 1) and lytic reagent (Reagent 2) and shutdown diluent (Reagent 3). Reagent 1 is an isotonic electrolyte solution that dilutes whole blood samples, stabilizes cell membranes for accurate counting and sizing, conducts aperture current, rinses instrument components between analyses, and prevents duplicative cell counts by using the sweep-flow process. Reagent 2 is a lytic reagent that lyses red blood cells for white blood cell count and hemoglobin measurement. Caution: Eye irritant. Avoid contact with skin and eyes. Avoid breathing gas.
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Contact with acid liberates poisonous gas. Reagent 3 is a shutdown diluent that prevents protein buildup that occurs in and around the apertures. Caution: Avoid eye and skin contact. Do not ingest. Reagent Preparation No reagent preparation is required.
Appropriate safety precautions for handling reagents are contained on the respective Safety Data Sheets located in the laboratory’s SDS/ HAZARDS binder. Reagent Storage Store reagents at ambient room temperature (2-25 C). Keep containers closed.
Discard reagents at the expiration date. Replace reagents when the screen prompt appears or when the reagent container is empty.
Reagent Tracking When opening new reagents, log on the Reagent Log indicating date opened, lot number, and expiration date. QUALITY CONTROL PROCEDURE QC Frequency Test 3 levels of QC samples once per day of testing using Coulter 4C PLUS cell controls. Order controls in Harvest. QC Procedure Using COULTER 4C PLUS Cell Control.
Ensure that 4C PLUS cell control is not past its expiration date and that it is at the correct storage temperature. Allow refrigerated controls/calibrators to sit 10-15 minutes at room temperature before use.
Mix by rolling slowly between the palms of the hands 8 times then invert and roll 8 times. Gently invert the tube 8 times.
Inspect the tube contents to determine if all cells have been uniformly distributed. Repeat above steps if tube contents have not been uniformly distributed Inspect the tube contents to ensure that all cells are uniformly distributed; if not, repeat this step. At the Main screen, touch the QA icon.
At the QA screen, touch the 4C PLUS Run icon. Select the correct control level: L, N or H. The square darkens next to your selection. Make sure that the level of control you are testing matches the one selected (L, N or H). Invert the tube once or twice prior to cycling. Place the well-mixed sample in the tube holder at the Cap Pierce Station and close the door.
When the tube holder door opens, remove the vial and return it to the refrigerator as soon as possible. Results appear on the screen. Unless non-numeric results occur for one or more parameters, the control results are automatically stored in the Coulter data storage. If you choose, curent control normals and ranges may be entered into the Coulter. (See procedure in the Coulter Easy Reference Guide under Quality Control Set Up.) Since all results are dealt with in the LIS, abnormals appearing on the coulter screen are only there for interest sake. Click on the control you've just run in LIS. Any abnormals will appear in red. 1 2s errors will appear in light red.
Other errors - 2 2s, 1 3s and 10x, which suggest possible sytemic errors - will appear in dark red. Follow laboratory's protocol for troubleshooting out-of-range controls. See attached procedure: 'Troubleshooting Out-Of-Range Controls.' . If the results are within the expected range, you are finished running controls.
Hematology Sysmex Procedure Manual
If you do all of the above steps and the results still do not meet your performance expectations, inform your supervisor. If the supervisor is unavailable, call your Beckman Coulter Representative who can guide you in choosing approprate trouble shooting procedures in the Coulter Operator's Guide, or may schedule a sevice call.
Do not report patient test results until control values are acceptable. Sample Analysis - Closed Vial Whole Blood Mode. At the Main screen, select Closed Vial Whole Blood mode. At the Main screen, touch the Sample Results Screen icon. NOTE: If the door is inadvertently closed after it has opened automatically, or if it is closed at a screen where samples are not run, you can open the door by touching the Main Menu icon and then the Sample Results icon. Type in the sample ID assigned to specimens by the Orchard Harvest LIS. Mix the sample on the mechanical rocker prior to sampling.
Be sure you are in the Closed Vial Whole Blood mode. Check the sample for clots by tilting the tube back forth and observing for clots. Place the well-mixed sample in the tube holder at the Cap Pierce Station and close the door. When the tube holder door opens, remove the tube. Sample results are automatically saved by the instrument, and the results appear on the screen.
To print the results touch the print icon. Results are verified in the LIS. Sample Analysis - Open Vial Whole Blood Mode. At the Main screen, select Open Vial Whole Blood mode. At the Main screen, touch the Sample Results Screen icon. Type in the sample ID. Mix the sample thoroughly on the mechanical rocker.
Be sure you are in the Open Vial Whole Blood mode. Present the well-mixed sample to the probe so that the tip is well into the tube, and press the aspirate switch. When you hear the beep, remove the sample, and put the cap back on the tube. The analyzer displays the sample results on the screen and automatically saves them. To print the results, touch the print icon. Results are verified in the LIS.
Use the Operator's Guide for: Getting started and running the instrument day‑to‑day Reviewing unusual results (how to read a result report and what flags mean) Performing special procedures such as cleaning, replacing, or adjusting a component of the instrument Troubleshooting problems with your instrument. Use the Reference Manual for: What the instrument does and methods it uses Instrument specifications and requirements How to interface your analyzer to your laboratory's host computer How to safely use the instrument.
Use the Installation and Training Guide for: Initially setting up the instrument and printer Powering up the instrument Customizing the software. Use the Operating Summary for: Running your instrument using a quick reference set of procedures Verifying screen icon References Coulter AcT diff2 Analyzer Operator's Guide PN 4237495B (June 2003) Copyright Beckman Coulter 1999,2003.
. GENERAL INFORMATION A. Hematology Laboratory Hours: Routine Hours: Monday-Friday, 8 am- 5 pm Emergency (STAT) Service: 24 hours/7 days B.
Telephone Numbers: Hematology – Information and Results 206-8590. TEST CONSULTATION AND APPROVAL Some hematology tests are performed ONLY after consultation with a Laboratory Medicine Resident. See the for further information. A Laboratory Medicine Resident is available at all times for hematology and coagulation problems: Monday-Friday 8 am - 5 pm, pager (415) 443-0179 At all other times, call the Clinical Laboratory Information Section (Inquiry) at 415-206-8590, and request their assistance to contact the Lab Medicine Resident on call. REQUISITION All hematology specimens should be submitted on one of the following requisitions:.