1. Peripheral smear

2. Preparations Specimen-direct capillary blood or anticoagulated blood Apparatus-clean glass slides,lancet, spreader slide,microscope&microscope oil Stain-leishman stain,wright or gimsa Diluent-distilled water or buffer solution with Ph 6.8 3. Composition of leishmans stain Polychromed methylene blue Eosin azure leishman powder Acetone free absolute alcohol 1.5gms leishman powder in 1 liter acetone free absolute alcohol 4. Procedure MAKING A SMEAR One drop of blood on one end of slide Spreader slide is placed at 45on the drop and moved along the slide It is moved smoothly and once such that the blood film is thin Care should be taken to prevent the formation of air bubbles Air dry the smear Make identification mark on one edge 5. Staining a smear Place the smear on the staining rack Pour leishman stain to cover the smear completely allow to fix for 2-3 mins Add water twice the amount of leishmans stain allow to fix for 7-10 mins Appearance of golden scum or sheen on the surface of stain Wash the stain off the slide with running water Wipe the back of slide and air dry the slide 6. Examination of the smear The stained slide is placed on the microscope and is focused under low power Place a drop of oil on the slide and observe under oil immersion objective Observe RBC,platelet series WBC series is observed and a differential count is done Record the findings 7. Precautions Objective must be clean Objective must be cleaned after every use Good smear is tongue shaped occupying 2/3rd of the slide Divided into head,body and tail Must be focused at the junction of body and tail 8. Diagnostic value RBCs SERIES Size-microcytic/normocytic/macrocytic/ anisocytosis Shape-poikilocytosis,sickle cells,target cells,tear drop cells,burr cells spherocytosis,ovalocytosis Color-hypochromic/normochromic Immature cells- normoblasts,polychromatophillic cells 9. WBCs SERIESDIFFERENTIAL COUNT Neutrophills-40--70% Eosinophills-2--4 % Basophills-0--1 % Monocytes-2--4 % Lymphocytes-2540 % 10. Immature cells Myeloid series- myeloblasts,promyelocyte,myelocyte,met amyelocyte,band forms Lymphoid series-lymhoblasts 11. Thank you 12. PLATELET SERIES General score-6--10platelets/oil immersion field PARASITES Intracellular-malarial parasite Extracellular-microfilarial