Blood Cell Counts PDF

56 b. Sources of error 3. Calculations ❖ Cold Agglutinins ❖ High WBC Count Formula:...

56 b. Sources of error 3. Calculations ❖ Cold Agglutinins ❖ High WBC Count Formula: # cells counted x x dilution 4. Reference range - male: 47 ± 5% female: 42 ± 5% Tot vol* factor NOTE: Volume (of l square) = length x width x depth *Total volume = volume x # of squares c ounted a. WBC count ❖ Coimt all 9 squares ❖ Total volume counted = 1mm x 1mm x 0.1mm x 9 = 0.9mm3 ❖ Reference range - 5,000 - 10,000/ul b. Platelet count Sources of Error m - ❖ Count all 25 squares in center Hemogl.obiD. and Hematocrit large square TestiD.K ❖ Tota.I volume counted = 1mm x 1mm x 0.lmm'1 = 0.lmm3 MANUAL CELL COUNTS ❖ Reference range - 1. WBC/Platelet Count 150,000 - 400,000/ul a. Diluent - 1.98 ml of 1% Ammonium ❖ WBC example Oxalate b. 0. 02 ml or 20µ] pipet WBC's counted = 60 c. 1:100 dilution (0.02 ml in 2.00 ml) 60 x _1_ x 100 = 6,667/mm3 or µL.9 2. Hemacytometer (Neubauer) NOTE: You can eliminate the 1/. 9 (volume 1mm 1mm 1mm factor) by multiplying 10% of cells counted and adding this value to the number of cells counted. 1mm Example: 60 cells counted x 10% = 60 + 6 = 66 Now: 66 x 100 = 6,600/mm3 or µL 1mm ❖ Platelet example Platelets counted = 150 150 x _ I x 100 = 150,000/ mm3 orµL 1mm.1 NOTE: Multiply number of platelets counted by Depth of Chamber 0.1mm 1000. Red Cell Indices INDICES FORMULA REF. RANGE - - - - - INDICATION - - - - - - - MCV* HCT x 10 80-1 OOfl (µ3) 100 Macrocytes MCH HB x l0 28-32pg (µµg) Varies with Hemoglobin Content and Cell Size RBC HB x 100 36 Spherocytes, lcterus, Lipemia, Hb SS, CC ROW-CV SD of MCV x 100 11.5-1 4.5% >14.5 Anisocytosis Mean MCV Width of peak at 39-47 il Anisocytos is ROW-SD 20% height >47 fl I MCV is directly measured by automated methods; Hct is calculated from MCV and ABC count. I  57 Peripheral Blood Cells and Their Function CELL REFERENCE RANGE FUNCTION Red Cell Females 3.8 - 5.2 x 10"/~ I 0 2 Transport to Tissue and CO 2 Removal from Tissue Males 4.5 - 6.1 x 10"/µ Neutrophil Relative 45-70% lnj est and Kill Bacteria Absolute 2250-7000/µ l Lymphocyte Relative 20-40% H umoral and Cell Mediated Immunity Absolute 1000-4000/µ I Monocyte Relative 3-10% lnjest and Kill Bacteria, Digest Debris, Absolute 150-1000/µ 1 Initiate and Regulate Adaptive Immune Response Basophil Relative 0-2% Inflammatory Response Mediator Absolute 0-200/µI Eosinophil Relative 0 -3% Al lergic Response Regulator Abso lute 0-300/µ1 Platelet Absolute 150,000-400,000/µI Clotting BLOOD SMEAR EXAMINATION 1. Wright stain a. Polychrome (Romanowsky) stain - phosphate buffer (pH 6.4) , eosin, methylene blue and methanol (fixa- tive) b. Sources of error ❖ Stain too blue (pH of buffer or stain too basic, prolonged stain- c. Significance of increased leukocytes ing) CELL INCREASED IN ❖ Stain too red (pH of buffer or Neutrophil Bacterial Infections stain too acid, prolonged washing) Lymphocyte Viral Infections 2. Leukocyte differential a. Normal Relative(%) values - see Monocyte TB, Syphilis, Malignancies chart above Allergies, Parasites Eosinophil b. Absolute values ❖ Calculation Basophil Immediate Hypersensitivity Relative value (%) x Total lfl_BC count ❖ Example Total WBC count = 6000/uL d. ·w hite cell morphology % Neutrophils from diff = 60% MORPHOLOGY ASSOC IATED WITH Absolute neutrophil count= Hypersegmented Neutroph iI Megaloblastic Anemia 0.6 x 6000/uL = 3,600/uL (B 12 and Folate deficiency) Hyposegmented Neutrophil Pelger Huet , Pseudo-Pelger Huet (AML *, A IDS) Toxic Granulation and Bacterial Infections, Burns, Vacuoles Chemotherapy Di:ihle Bodies (RNA) Bacterial Infections, Burns, May-Hegglin Variant Lymphs (increased size Infectious Mono, Other Viral Wright Stain Components and basophilia) Infections * Acute Myeloid Leukemia 58 3. Platelet estimate ❖Hyperchromic (MCHC > 36) a. 8-20/oil immersion field- adequate ❖Polychromasia - blue color in rbc h. If platelets seem low, check feather (if stained with new methylene edge of slide for platelet clumping, blue, these would be reticulocytes) check for satelitism (EDTA related- c. Shape - Bi-concave disk redraw in Na citrate) ❖ Poikilocytosis - variation in shape c. Check platelet size (see chart below) ! Large - Bernard-Soulier, d. Inclusions ( see chart page 59) May- Hegglin, myeloprolifera.tfre e. Crystals disorders, stress platelets ❖ Hemoglobin C - bar-shaped ❖ Hemoglobin SC - "hand in glove" 4. Nucleated red cells or " Washington monument" a. Must correct white cell count (if included in count) b. Formula wbc count x 100 = corrected WBC count 100 + # nrbcs 5. Red cell morphology WBC Reference Ranges, Relative vs.Absolute Values, a. Size Cell Functions ❖ Normocytic (MCV 0f80-1()()µ 1 orfL) ❖ Microcytic (MCV < 80) ❖ Macrocytic (MCV > 100) Correlate Conditions witb ❖ Anisocytosis - variation in size Increased WBCs and Abnormal Morphology (RDW> 14.5) b. Color ❖ Normochromic (MCHC of Action Taken ifLow 32-36 gldL) Platelets Seen on Smear ❖ Hypochromic (MCHC < 32) Abnormal Red Cell Shapes - - SHAPE SEEN IN: SHAPE SEEN IN: ~ p Abetalipoproteinemia, Severe Sickle Cell (Drepanocyte) Hb SS Liver Disease E,h;'ljcell Uremia, Artifact Spherocyte Hereditary Spheroitosis, (Alkaline Class Effect) (+ MCHQ, ABO H N, and 0 Other Hemolytic Processes Elliptocyte Hereditary Elliptocytosis, Iron Stomatocyte (Mouth Cell) Hereditary Stomatocytosis Deficiency, Thalassemia Liver Disease (0 8 Liver Disease, Hb C, Macroovalocyte Megaloblastic Anemia Target Cell (Codocrte) Thalassemias, and Other Hemoglobinopathies ~ @) Hemolytic Processes Teardrop Cell (Dacryocyte) Extramedullary Hematopoiesis, HSm~ocytel D Thalassemias, Pernicious Anemia Schistocyte (RSC Fragment) DIC and Hemolytic Processes NOTE: HD 1 - Hemolytic Disease of the Newborn % DIC - Disseminated lntravascular Coagulation)  59 Red Cell Inclusions INCLUSION COMPOSED OF: STAIN INDICATIONS Howell-Jolly Body DNA Wright Disturbed Erythropoiesis New Methylene Blue Hemolytic Anemias 0 Megaloblastic Anemia Post-Splenectomy Basophilic Stippling RNA Wright Thalassemia.......... New Methylene Blue Lead Poisoning. :·;:: : ~- :... ·: ·:t...... I · ♦ ♦ 0 Pappenheimer Bodies Iron Wright Sideroblastic Anemia Siderotic Granules (siderocyte) Confirm with Prussian Blue Hemoglobi nopath ies 0 Heinz Body Denatured Precipitated Supravital Stain G6PD Deficiency Hemoglobin such as Brilliant Cresyl Blue or Thalassemia 0 New Methylene Blue Unstable Hemoglobins (NOT seen with Wright stain) g Cabot Ring Remnants of Mitotic \.\/right Megaloblastic Anemia Spindle Parasites Malaria Wright Parasitic Infection Babesia Trypanosomes NOTE: ~ ,i. MCV, ,i. MCHC and t red cell count are associated w ith cold agglutinin disease; warm sample and rerun Correlate AbnontuullBC Shapes with Conditions Correlate-Abnormal RBG Inclusions with REMEMBER! HEME NO RMALS Conditions RULE OF THREE 9 d' Correlation of Hb, Hct and RBC Hb 14 ± 2 g/dl 16 ± 2 g/dL Hb x 3 = Hct ± 3% Hct 42 ± 5% 47 ± 5% RBC (in millions} X 3 = Hb ± 0.5 6 RBC 4.5 ± 0.7xl 0 /µL 5.3 ± 0.8x1 0'/µL If "Rule of Three" doesn't fit, consider: 7. Clotted sample WBC 5,000 - 10,000/µL 2. Cold agglutinin (warm sample and rerun) 3. Lipemic, icteric, or grossly hemolyzed sample Platelet 150,000 - 400,000/µL 4. Resisting hgb (SS, CC) 5. Abnormal red cells