Beckman Coulter AcT diff2 Series Analyzer Procedure …

1 1 Beckman Coulter AcT diff2 Series Analyzer Procedure manual Cowell Student Health Center Laboratory, 2006 2 Policies and procedures Approval Form Procedure : AcT diff 2 Series Analyzer Prepared By: _____ _____ Name Title Approved By: _____ Laboratory Director Date Adopted: _____ Date Discontinued: _____ Read By: Date Name/Title Revisions: Review Date Revision Date Approved By Title 3 Personnel who actually perform moderately complex tests ( , the AcT diff2 Series Analyzer ) are referred to as testing analysts by CLIA and must have at least a high school diploma.

2 Testing personnel must have documented training and undergo periodic competency evaluations. In this laboratory, the following persons fulfill the role of: Laboratory director:_____ Clinical consultant: _____ Technical consultant: _____ Testing analyst(s): _____ _____ In this laboratory, the following personnel are authorized to operate the AcT diff2 Series Analyzer : Employee Name Employee Title Supervision Required? Y/N Sup. Review to Report Results? Y/N 4 Principle The Coulter AcT diff2 Series Analyzer is a quantitative, automated hematology Analyzer and leukocyte differential counter for In Vitro Diagnostic Use in clinical laboratories.

3 AcT diff2 Series Analyzer parameters Parameter WBC - White Blood Cell or leukocyte count LY# Lymphocyte number LY% Lymphocyte percent (or ratio) MO# Mononuclear cell number MO% Mononuclear cell percent (or ratio) GR# Granulocyte number GR% Granulocyte percent (or ratio) RBC - Red Blood Cell or erythrocyte count Hgb - Hemoglobin concentration Hct - Hematocrit (relative volume of erythrocytes) MCV - Mean Corpuscular (erythrocyte) Volume MCH - Mean Corpuscular (erythrocyte) Hemoglobin MCHC - Mean Corpuscular (erythrocyte) Hemoglobin Concentration Plt - Platelet or thrombocyte count RDW - Red Cell (erythrocyte volume) Distribution Width MPV - Mean Platelet (thrombocyte) Volume The purpose of the AcT diff2 Series Analyzer is to identify the normal human patient, with all normal system-generated parameters, and to flag or identify patient results that require additional studies.

4 AcT diff2 analyzers are based on the Coulter method whereby cells are counted and sized by detecting and measuring changes in electrical resistance when a particle (such as a cell) in a conductive liquid passes through a small aperture. As each cell goes through the aperture, it impedes the current and causes a measurable pulse. The number of pulses signals the number of particles. The height of each pulse is proportional to the volume of that particle. While the number of pulses indicates particle count, the amplitude of the electrical pulse produced is proportional to the cell s volume. The AcT diff2 Analyzer has two operating modes: Open Vial Whole Blood and Closed Vial Whole Blood.

5 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 K2 EDTA. Thoroughly mix blood with EDTA anticoagulant. If hemolysis or small clots are observed, discard specimen. 5 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.

6 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- 8C. Warm samples to room temperature (16-35C or 61-95F) 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 pack or diff AcT Pak , both of which contain diluent (Reagent 1) and lytic reagent (Reagent 2).

7 The diff AcT Tainer reagent pack also contains AcT Rinse 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. Contact with acid liberates poisonous gas. Reagent 3, AcT Rinse Shutdown Diluent, prevents protein buildup that occurs in and around the apertures.

8 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 Material Safety Data Sheets located in the laboratory s MSDS/ HAZARDS binder. Reagent Storage Store reagents at ambient room temperature (2-25C). 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. Coulter 4C PLUS cell control: abnormal low, normal and abnormal high.

9 Coulter S-CAL calibrator. 6 Control/Calibrator Preparation Handle controls and calibrators using universal precautions. Controls and calibrator contain stabilized human erythrocytes and no test method can offer complete assurance that Human Immunodeficiency Virus (HIV), Hepatitis C Virus (HCV), Hepatitis B Virus (HBV) or other infectious agents are absent. Take appropriate safety measures to avoid contact from aerosols when removing the cap/stopper assembly. Control/Calibrator Storage Store at 2-8C (35-46F). Sealed control and calibrator vials are stable until the expiration date. Opened calibrator vials are stable for 1 hour.

10 Discard expired control/calibration materials. Before use, inspect controls/calibrators for indications of instability or deterioration. Gross hemolysis (darkly colored supernatant) is indicative of product deterioration. Do not use deteriorated product. Waste Container Be sure the waste container is in a safe place and is properly connected. Do not overfill the waste container. When the "waste full" icon appears, replace the waste container with an empty one. Quality Control procedures QC Frequency Test 3 levels of QC samples once per day of testing using Coulter 4C PLUS cell controls. QC Procedure Using Coulter 4C PLUS Cell Control 1.