Bluegrass Community HospitalLaboratory

360 Amsden AvenueVersailles, Kentucky 40383

Procedure Description: Peripheral Smear Review

Prepared by:

_______________________________ ______Norma J. Whitworth, BSMT(ASCP) DateLaboratory Manager

Approved by:

________________________________ _______Richard Lozano, M.D. DateMedical Director

Effective Date: Revised:

Annual Review:

Principle: The Cell-Dyn 3200 uses two independent measurement channels to measure, count, and calculate hematological parameters. The Optical channel uses flow cytometry to determine the WBC, NOC, and RBC/PLT data. The Hemoglobin channel uses colorimetric determination for hemoglobin. During each instrument cycle, the sample is aspirated, diluted, and mixed before each parameter is measured. The Cell-Dyn 3200 monitors instrument conditions and data criteria that may affect results and displays appropriate operational messages and flags for operator review. Certain messages and flags prompt the lab technologist to perform a ‘SCAN” or “Manual Differential (Man Diff)” of a peripheral blood smear. Findings after a “SCAN” may designate a manual differential to be performed. The following protocol designates “SCAN” and “Man Diff” criteria. Precautions: Blood and body fluids should always be tested using Universal Precautions. Gloves, spill-proof gown, and protective eyewear may be needed. Reagents used for CBC and individual parameters are not for invitro testing. Refer to each reagents MSDS sheet for specific information regarding handling and hazards.

Storage and Stability: Hematology specimens and peripheral smears are kept for 7 days before discarding. Whole Blood EDTA specimens are stored at 2 – 8 C. Cell-Dyn 3200 reagents are stored at room temperature.

Specimen Collection and Handling:

A 3 CC EDTA whole blood specimen is required. Refer to the Blood Smear Preparation procedure in the Hematology SOP for details are preparing a peripheral blood smear for scan or differential review.

Materials Needed: Cell-Dyn 3200EDTA vacutainer tubeFrosted slideDiff-safe segment or heparinized capillary tubeWright StainMicroscopeOil Immersion

Quality Control: Peripheral smear review policy is reviewed annually and approved by MedExec. Technologists are evaluated annually and quarterly on proficiency specimens.

Procedure: 1. Sample a 3 cc EDTA whole blood specimen with the Cell-Dyn 3200. 2. Review histogram for any interpretative messages and/or flags. Refer to the Cell-Dyn

3200 Operator’s Manual for detail discussion of histograms.3. Refer to the chart below to evaluate results for the need for a peripheral smear scan or

differential.

TEST FLAG/MESSAGE ACTIONCBC with DifferentialCBC without Differential

NWBC Scan: review for platelet clumps, giant platelets, or NRBC’s. If no other flags or criteria for manual diff present, report automated WBC and diff.

NRBC Scan: review smear for NRBC’s.

WBC Confirm WBC by Unopette method.

FWBC Repeat CBC test in FWBC mode. The NOC result will be reported for the WBC.

VAR LYMPH or VAR LYM Scan: review smear to confirm lymph count, perform a manual diff if variant/atypical lymphs are noted.

DFLT Perform a manual differential.

IG (immature granulocytes) Perform a manual differential.

TEST FLAG/MESSAGE ACTION

CBC with DifferentialCBC without Differential

BLAST Perform a manual differential.

BAND Scan: review smear for bands, perform a manual diff if indicated.

RBC MORPH

Flag is triggered if the following limits are exceeded:MCV < 80 or > 100 fLMCH < 25 or > 34 pgMCHC < 29 or > 37 g/dLRDW > 18.5 %

Perform a RBC morphology only. Indicate platelet morphology and platelet estimate also.

CBC with DifferentialCBC without DifferentialH&H

RRBC Repeat CBC test in RRBC mode. If flag persist, scan smear for NRBC’s.

CBC with DifferentialCBC without DifferentialPlatelet Count

LRICause: debris, contaminated reagent, microbubbles, dirty diluent/sheath filter

Repeat Platelet count. If flag persists, review smear and perform a Platelet estimate and morphology.

Verify platelet background if problem noted on subsequent specimens.

URICause: Microcytic RBC’s, schistocytes, giant platelets, sickle cells, platelet clumps

Repeat Platelet count and review smear if flag persists. Perform a Platelet estimate and morphology.

LURI Repeat Platelet count. If flag persists, review smear and perform a Platelet estimate and morphology.

CBC with Manual Differential

Manual Diff required.

WBC < 2.0 or >20.0 K/uL Manual Diff

15.0 – 19.9 K/uL Scan: review smear for immature granulocytes or other diagnostic parameters, perform a manual diff.

Lymphocytes > 60.0 % Manual Diff

Monocytes > 15 % Manual Diff

Neutrophil Absolute Count > 10.0 Manual Diff

Lymphocyte Absolute Count > 8.0 Manual DiffTEST FLAG/MESSAGE ACTION

Hemoglobin (HGB) < 8.0 or > 18.0 g/dL RBC Morphology

Hematocrit (HCT) < 24.0 or > 54.0 % RBC Morphology

MCV < 80.0 or > 100.0 fL RBC Morphology

Platelet Count < 100 or > 800 K/uL Platelet Morphology and Estimate

RDW > 18.5 % RBC Morphology

Results and Interpretation: A manual differential, scan, RBC morphology, or Platelet morphology will be performed when any of the above stated criteria are met or the lab technician or physician suspects a clinical state. Any result rechecked should be designated by an “*” with the comment “Results repeated and verified.”

References: 1. Abbott Diagnostics: Cell-Dyn System 3200 Operator’s Manual, Abbott Park, IL, 60064,

2001-2002.