Hematology 101 Cindy Rogers, MT(ASCP) Diagnostics System Specialist
More Acronyms...» CBC» RBC» HGB» HCT» WBC» MPV» PLT» RDW» DIFF» H&H» Complete Blood Count» Red Blood Cell» Hemoglobin» Hematocrit» White Blood Cell» Mean Platelet Volume» Platelet» Red Cell Distribution Width» WBC Differential» Hemoglobin and Hematocrit
The Objec)ves» The Basics of Hematology» Cell Lines» BCI Cell Counting and Differential Technology-VCS» Quality Control Basics» Calibration Basics» Q & A
What is Hematology? Greek: Heme (Blood) -ology(study of) The study of the cellular components of blood and blood producing organs. Consists of counting these cellular components. The information gathered from the CBC can assist clinicians in diagnosis/treatments of different conditions & disease states.
Composi)on of Blood Plasma: 55% Water (90%) Ions Nutrients Clo0ng Proteins An8bodies Hormones Buffy Coat 1% White Blood Cells (WBC) Platelets (PLT) Red Blood Cells (RBC) 44%
Types of Blood Cells Red Blood Cells Platelets WBC Blood Cells
Complete Blood Count (CBC) RED CELL PARAMETERS WHITE CELL PARAMETERS ØWhite Blood Cell Count ØWhite Cell Differential Neutrophil Lymphocyte Monocyte Eosinophil Basophil Ø Red Blood Cell Count Ø Hemoglobin Ø Hematocrit Ø RDW Ø Indices (MCV, MCH, MCHC) Ø Reticulocyte Ø Nucleated Red Blood Cells (NRBC) PLATELET PARAMETERS Ø Platelet Count Ø Mean Platelet Volume Ø Platelet Distribution Width
Red Blood Cell (RBC)also called Erythrocytes Red Blood Cell Count (RBC): number of cells per volume of whole blood Normal Ranges Adult Males: 4.8 ± 0.8 x10^6/ul Adult Females: 4.3 ± 0.6 x10^6/ul Primary Function: Transport Oxygen to the tissues/cells throughout the body via Hemoglobin
Hemoglobin (Hgb) Concentra)on Transport of Oxygen Main component of mature Red Cell (=1/3) Hgb gives blood its red color Heme Molecule Normal Values (g/dl) Adult Males: 14.3 ± 2 Adult Females: 12.6 ± 2
Hematocrit (Hct) % by Volume of Packed RBCs. Calculated on DxH (RBC x MCV)/10 Adult males: 41.3 ± 5 Adult females: 36.9 ± 5 Spun Crit:
Hematocrit Normal è ç Anemia
Significance of Abnormal Values» Decreased: Anemia blood loss hemolysis deficiency of iron, vitamins B6 (folate), B12 Expanded plasma volume (e.g. pregnancy)» Above normal: Polycythemia Severe dehydration (e.g. burns) Polycythemia vera
Red Cell Indices MCV= Hct (g/dl) x 10 RBC (10 6 /ul) MCH = Hgb (g/dl) x 10 RBC (10 6 /ul) MCHC = Hgb (g/dl) x 100 Hct (%)
DxH Indices Formula for MCV= Hgb (g/dl) x 10 RBC (10 6 /ul) DxH HCT= RBC x MCV DxH MCV = Derived from the RBC Histogram
NRBCs or Nucleated Red Blood Cells» In the past NRBCs were enumerated when performing a manual diff (reported as #/100 WBCs)» NRBCs are normally found in the bone marrow» Because NRBCs are nucleated and larger than RBCs they can sometimes be counted as WBCs so the WBC count had to be manually corrected for NRBCs» Now analyzers are able to accurately enumerate NRBCs and correct the WBC NRBC
Re)culocytes (Re)c)» Immature RBCs with retained RNA in the cytoplasm that were previously NRBCs in the bone marrow» Reticulocytes circulate in the bloodstream for about 2 days before developing into mature red blood cells» Normally about 1-2% of RBCs are retics» The reticulocyte count provides information about the rate that the bone marrow produces RBCs Wrights Stain New Methylene Blue
Abnormal Re)culocyte Counts éacute blood loss or surgery Anemia ê Chronic Leukemia Metastatic Carcinoma If RBC production is inadequate, a retic count is performed. (bone marrow insufficiency vs. bleeding or RBC destruction) Peripheral blood smear Stained Reticulocyte smear
Platelet (PLT) Also called a Thrombocyte Cellular Fragment From Megakaryocyte in bone marrow Forms a Hemostatic plug to aid in Coagulation
Platelet Abnormali)es THROMBOCYTOPENIA (decreased platelets) Some anemias (pernicious) Acute leukemia Certain drugs and radiation Disseminated Intravascular Coagulation THROMBOCYTOSIS (increased platelets) Acute blood loss or surgery Polycythemia Vera Cancer Chronic Kidney Failure Some anemias (iron deficiency)
White Blood Cell (WBC) also called Leukocytes» Assist in fighting infection» Normal range in both males and females 7.7±3.3x10 3 /ul» 5 Basic Types are present
White Blood Cells (WBC) ê LEUKOPENIA Viral infections Endocrine disorders, Drugs & Radiation é LEUKOCYTOSIS Bacterial infections Acute Inflammatory Conditions Neoplastic disease (e.g. Leukemia)
WBC Differen)al» The WBCs are counted and then the types of WBC are enumerated» Each type of WBC has a unique function, but all are associated with providing immunity or defense from infection» Prior to analyzers with automated differential capability this was performed by the technologist using a stained blood smear and a microscope
Differen)al Results Relative Numbers Expected Neutrophils 50% to 75% Lymphocytes 20% to 45% Monocytes 3% to 11% Eosinophils 1% to 3% Basophils 0% to 1%
Abnormal Condi)ons Blasts seen in various Leukemias Bands or immature cells indicating Infection or leukemia (Left Shift or IG (Immature Granulocytes) Atypical lymphs as seen in Mononucleosis
The Coulter Principle Vacuum Aperture Current Pathway Internal Electrode Detail of Aperture External Electrode Suspension of Cells External Housing (Aperture Bath) Aperture Aperture Housing
Sizing the Cells The larger the cell, the more electrical current it blocks. The height of the pulse is equal to the volume of the cell.
VCSn Differential Development The Hydrodynamically Focused Flow Cell organizes the 8192 near native state cells into single file for analysis FLOW CELL SHEATH-FLUID IN SAMPLE DILUTION SENSING AREA SHEATH FLUID SHEATH-FLUID IN SAMPLE IN
Advanced Analytical Measurements» Seven parameter measurements 1. Volume Cell size, based on Coulter principle 1. Conductivity / Opacity - Internal characteristics, nucleus to cytoplasm ratio, nuclear density, chemical composition
Advanced Analytical Measurements» 5 angles of light scatter 3. MALS - Cell complexity, granularity, nucleus structure 4. AL2 0 degree light scatter for cell size based on absorbed light 5. LALS 6. LMALS 7. UMALS UMALS MALS LMALS LALS AL2 LALS MALS LMALS UMALS
DIFF NORMAL DATAPLOTS Mono Neut Eo NRBC Non-White WBC Lymph Baso Non-White RETIC Mature RBC WBC NRBC PLT Immature RBC / Retics
Abnormal Cell Detection 1 Mono-Blasts 2 Myelo-Blasts 2 1 3 Immature 3 Granulocytes 1 Mono-Blasts 4 4 Left Shift 2 Myelo-Blasts 5 Lympho-Blasts 3 Immature Granulocytes 6 Variant Lymphocytes 4 Left Shift 5 7 Low Volume Lymphocytes 6 5 Lympho-Blasts 8 Non-White Cells 6 Variant Lymphocytes 7 Low Volume Lymphocytes 7 8 Non-White Cells 8
Automated Differen)al vs. Manual Differen)al» Analyzers are designed to count normal cells and do an excellent job» Analyzers are also designed to notify the tech of the possibility of abnormal cells (Suspect Messages)» Each lab also has their own protocol for performing a manual diff when cells are present in certain numbers (Definitive Messages)» Techs perform manual diffs on those abnormal specimens
Review of Terms» CBC Complete Blood Count» Erythrocyte Red Blood Cell (RBC).» Leukocyte White Blood Cell (WBC)» Thrombocyte Platelet (Plt)» Hematocrit Percent of RBCs in whole blood. (Hct)» Hemoglobin- Weight of oxygen carrying substance in RBCs, a primary indicator of anemia.» MCV Mean Corpuscular (Cellular) ( Volume. Size of RBC (macrocytic, normocytic or microcytic).» MCH and MCHC Mean Corpuscular Hemoglobin and Concentration. Amount of hemoglobin in each cell, (Hyperchromic, normochromic, hypochromic).» NRBC- Nucleated RBC
Review of Hematologic Condi)ons» Leukocytosis» Leukopenia» Thrombocytosis» Thrombocytopenia» Polycythemia» Anemia» Granulocytosis» Left Shift» Immature Granulocytes (IG)» Agranulocytosis (Hypogranulation) Lymphocytosis Lymphopenia Atypical Lymphocytes Eosinophilia Basophilia Monocytosis Aplastic Anemia Leukemia Hemoglobinopathy Normal
Calibra)on» Calibration is a means by which we ensure that an analyzer is providing accurate results» A calibrator is a Hematology product consisting of a substance intended to establish points of reference (i.e., calibrate) of known values.» 6 parameters Calibrated WBC MCV RBC PLT HGB MPV
Calibra)on Frequency» Initial Install» Replacement of components involved in Dilution preparation Primary measurement» When advised to do so by your BCI Representative
When to Verify Calibra)on» Per laboratory procedures, local or national regulations (minimum of every 6 months per CAP)» Controls Show evidence of unusual trends/shifts Exceed manufacturer s defined limits» Room temp > 10 o F from calibrating temp
Quality Control» Reagents consisting of substances of known concentrations that are assayed by the usual clinical laboratory methods and that resemble the unknown specimen.» Principle: tests system failures and use errors that would negatively influence the testing of patient samples and will also affect the results obtained with the QC samples.» Required by regulatory agencies» Quality Control material such as 5C (LH series), 6C DxH series are stabilized blood products used to assess the performance of a Hematology analyzer
What is IQAP? Intra laboratory Quality Assurance Program» IQAP is BCI s program for assessing each lab s comparison to it s peers» All month the lab is trying to recover results close to the target values, but their comparison to peers is more important» The pool mean is considered the true value
Common Questions
What is a Delta Check?» Delta checking is a way for a lab to monitor changes in a particular patient s results» Usually defined as a % or absolute change» Utilization of delta checks by a lab can lead their doctors to make appropriate changes in patient s treatments Identify erroneous results > Wrong patient, diluted sample (MCV is a big one)
What does PRN mean?» A PRN is the acronym of the Latin word pro re nata which means as the situation demands. A term commonly used with nursing staffs, many Lab Directors will use this term when speaking about their own staff. A PRN is someone who will fill in when the laboratory staff is short on an as needed basis.
Your Questions??????