KHMC Hematological Tests and Bone Marrow Examination Department

KHMC Hematological Tests and Bone Marrow Examination Department of Laboratory Medicine Woo-In Lee

Hematological Tests & Bone Marrow Examinations Hematological Tests • Anticoagulants • CBC with Differential count • Reticulocyte • ESR • PBS Bone Marrow Examinations

Hematological Tests I. Anticoagulants • Tri-sodium citrate • • • EDTA • • • calcium chelating PT, a. PTT, Platelet aggregation test 항 응고제 1 : 혈액 9의 비율로 채혈 Hematocrit에 따른 항 응고제 비율 조정 [1: 5 ~ 1: 6~~~~1: 9(45%) ~ 1: 11] Calcium chelating Blood cell count, cell morphology Heparin • • Anti-thrombin III 와 결합하여 thrombin 억제 GAS analysis, osmotic fragility test

Hematological Tests II. Complete Blood Cell Count (CBC) CBC Traditional Unit International Unit Hemoglobin Hematocrit g/d. L (12~16) d. L/d. L(%) (45%) g/d. L L/L(%) Red Blood Cells(RBC) 106/u. L (3. 8 ~ 5. 4) 1012/L White Blood Cells 103/u. L (4. 0 ~ 10. 0) Platelet count 103/u. L (100 ~ 400) 109/L

Hematological Tests CBC RBC index Formula Traditional * International Unit MCV (평균용적) Hct(%) x 10 Fl Mean corpuscular volume MCH (평균혈색소) Mean corpuscular Hb MCHC (평균 혈색소농도) Mean corpuscular Hb conc. RDW (크기분포의 변이계수) Red Cell distribution Width RBC count(106/u. L) Hb(g/d. L) x 10 RBC count(106/u. L) Hb x 100 Hct SD x 100 MCV pg g/d. L * g/L

Hematological Tests WBC Differential Count(D/C)-백혈구감별계산 • Automated or manual • Granulocyte – Segmented neutrophil – Eosinophil – Basophil • Lymphocyte • Monocyte

Hematological Tests Platelet (혈소판) • 혈소판수 – 검체의 부적절한 상태에 따라 실제보다 높게 혹은 낮게 측정 – Pseudothrombocytopenia – Ex) EDTA - induced pseudothrombocytopenia • 말초혈액도말검사 • Citrate 항응고제 이용하여 다시 측정 • Mean platelet volume(MPV), platelet distribution width(PDW) • Reticulated platelet – 골수의 보상적인 혈소판생성능력 예측

Hematological Tests • Corrected Reticulocyte Count = Reticulocyte count x patient Hct 정상 Hct(0. 45) • Reticulocyte Production Index = Corrected reticulocyte / maturation time ** Hct 에 따른 maturation time (days) 45% - 1. 0, 35% - 1. 5, 25% - 2. 0, 15% - 2. 5

Hematological Tests Normoblast Reticulocyte Bone Marrow PB Wright stain - polychromasia Supravital stain - reticulocyte RBC

Hematological Tests IV. Erythrocyte Sedimentation Rate(ESR) • Principle • Factor – 혈장인자 • Fibrinogen, alpha 2, beta, gamma globulin zeta potential 저하 Rouleaux formation(연전) ESR 증가 • Albumin, lecithin은 ESR 감소 – 적혈구인자 • 빈혈, 적혈구 크기, 모양 • Interpretation – 0 ~ 9 mm/hr(M), 0 ~ 20 mm/hr(F) – 감염, 염증성 질환 등의 활동성 정도

Hematological Tests V. Peripheral Blood Smears (PBS) • CBC 검사 결과를 추정 – Hb, Hct, RBC index (MCV, MCHC, RDW) – WBC count • 200배 시야 field 당 평균 WBC No. x 1, 000/μL – Platelet count • 1, 000배 시야 field 당 평균 혈소판 수 x 20, 000/ μL – Reticulocyte count : polychromasia • 각 cell의 shape, size, 분포 등을 관찰 – PBS 의 ideal zone 에서 판독

Hematological Tests Ideal zone

Hematological Tests • 적혈구 크기의 이상 (Anisocytosis) – normocytic, microcytic, macrocytic • 적혈구 염색성의 이상(Hemoglobin 양) – normochromic, hyperchromic Normocytic normochromic Microcytic hypochromic Macrocytic hyperchromic Dimorphic normochromic

Hematological Tests • 적혈구 구조의 이상 Howell Jolly Body Basophilic Stippling Megaloblastic anemia Splenectomy Iron deficiency anemia 적혈구 생성 증가 Normoblast Malaria infection & Stippling(Schuffner’s granules)

Hematological Tests • 적혈구 모양(형태)의 이상 (Poikilocytosis) Elliptocyte Iron deficiency anemia 정상 성인 10% Spherocyte Cong. Spherocytosis Autoimmune hemolytic anemia Target cell Schistocytes (fragmented cell) Hemolysis (MAHA) Burn Megaloblastic anemia DIC Acanthocytes Abetalipoproteinemia Liver Disease Crenated cell Artifact DDx. Burr cell(kidney dis) 폐쇄성 황달 비장 절제술 후 Tear drop cell Sickle cell Myelofibrosis Sickle cell anemia

Hematological Tests • 기타 Rouleaux formation Platelet clumping Agglutination Giant Platelet

Bone Marrow Study Bone Marrow Examination • Indication • Bone Marrow Aspiration & Biopsy • Stain – Wright, Giemsa, Prussian blue(iron) – Reticulin(fibrosis), etc

Bone Marrow Study • BM Aspiration 관찰 – – – 세포의 형태, 감별계산, 비정상세포, etc Erythroid precursors (normoblast) Myeloid series Megakaryocytic series M: E ratio 정상 2 ~ 4 : 1 기타

Bone Marrow Study Hematopoietic Stem Cell Myeloid Stem Cell CFU-GM Lymphoid Stem Cell BFU-E CFU-Meg Monoblast Myeloblast Pronormoblast Megakaryoblast Pro. Monocyte Promyelocyte B. Normoblast Promegakaryocyte Megakaryocyte Neut. Myelocyte Eosin. Myelocyte Baso. Myleocyte P. Normoblast N. Metamyelocyte E. Metamyelocyte B. Metamyelocyte Ortho Normoblast Band neutrophil Monocyte Segmented Neutrophil B, T Lymphoblast Reticulocyte Eosinophil Basophil Erythrocyte Platelet Lymphocyte B, T

BM Myeloblast (Monoblast) Promormoblast Megakaryoblast Promyelocyte (Promonocyte) Metamyelocyte (N, E, B) Hematologic Malignancies Shift to left maturation PB Myelocyte (N, E, B) Megakarycyte Leukoerythoblastic Reaction Lymphoblast

Bone Marrow Study • BM Biopsy 관찰 • Cellularity (세포충실도) • 세포 분포 • 비정상 세포 유무 • Tumor invasion • Granuloma • Stage Work-up • 기타