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Blood Smear preparation.pdf
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- 2. ✿ To studymorphology of rbc ✿ To study morphology of wbc ✿ To study morphology of platlets ✿ To study morphology of inclusion bodies ✿ To confirm diagnosis of blood cell disorders ✿ To examine hemoparasites(malaria, trypanosoma etc..)
- 3. TYPES OF BLOODSMEAR ✿ Thin blood smear ✿ Thick blood smear
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- 5. THIN BLOOD SMEAR ✿This can be done achieved by anticoagulated blood... either obtained by venipuncture or finger prick ✿ There are 3 methods: ✿ 1) Slide method ✿ 2)cover glass/cover slip method ✿ 3) Spin method
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- 7. PROCEDURE ✿ Take agrease free, clean and dry slide ✿ place a drop of blood 1cm away from the end of the slide ✿ Keep the spreader slide infront of an <gle og 30-45° to the slide ✿ Draw the spreader backwards until it touches the blood ✿ Then blood will be flowing along the edge of the spreader slide ✿ Keep the <gle of spreader constant and push it front wards ✿ Dry it stain it and observe
- 12. PARTS OF THINBLOOD SMEAR ✿ They have 3 parts ✿ 1)Head ✿ 2)Body ✿ 3)Tail
- 14. QUALITIES OF AIDEAL BLOOD SMEAR
- 16. CAUSES OF POORBLOOD SMAER ✿ Drop of blood too large or too small. ✿ Spreader slide pushed across the horizontal slide in a jerky manner. ✿ Failure to keep the entire edge of the spreader slide against the horizontal slide while making the smear. ✿ Failure in using appropriate angle for the spreader slide. ✿ Failure to push the spreader slide compleyely acrosss the slide ✿ Holes in film slide contaminated with fat or grease ✿ Cellular degenerative changes delay in fixing inadequate fixing time or methanol contaminated with water
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- 26. STAINING OF THINBLOOD SMEAR
- 27. METHOD ✿ Leishman's staining ✿Wright's stain
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- 29. WRIGHT'S STAIN: DIPMETHOD
- 30. WRIGHT'S STAIN: RACKMETHOD
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- 32. ✿ It isused for detection of blood parasites ✿ For differential count ✿ In case of severe leucopenia
- 33. METHOD 1:BLOOD SMEARPREP ✿ Thick blood smear are prepared by taking 4 drops of blood on a slide ✿ Joining the corners of the drop with a needle or with the cornerof another slide ✿ Spread the drop in one direction to make a thick film, size of a icon diamter ✿ The thickness of the film should be such that it allows a mews print to be tead through dry prep
- 34. METHOD 1: DEHAEMOGLOBULIZATION ✿Dry the smear, dip in a copling jar containing 10drops pf glacial acetic acid and 50ml normal saline ✿ Repeat process till dehemoglobulisation (slide is colour less)is complete ✿ Drain it completely and wash with normal saline ✿ Stain smear with dil Gemsa or Leishman stain
- 35. METHOD 2 ✿ preparesmear ✿ Without drying immediatly put in 95% ethyl alcohol or 85% isopropyl alochol for 15 min ✿ When fixation is complete take out and pour few drops of glacial acetic acid ✿ Wait for few sec ✿ When dehemoglobulisation is complete througly wash with water
- 36. METHOD 2: STAINING ✿Fields stain A ✿ Fields stain B ✿ Two contaniner of clean water ✿ Air dry, dip into stainA fpr 5 sec drain ✿ Wash for abt 5 sec on watet drain off ✿ dip in to stain B for 5 sec drain off ✿ Wash with water gently, dry it and observe
- 37. EXAMINATION OF UNSTAINEDSMEAR ✿ 2-3 drops of blood is taken in a slide and coverslip us put on the rim is then smear with vaseline to prevent drying up of the blood ✿ Slid is examined under low power, if microfilaria is present shows wriggling movement ✿ Also used to identify trypanosomes ✿ Advantages ✿ Simple and inexpensive ✿ If trypanosomes is detected disease is diagnosed
- 39. BUFFY COAT PREPRATION ✿It is well known that a typical or primitive blood cell circulate in a small no in the peripheral blood in health ✿ A typical mononuclear cells, metamyelocyte and megakaryocytes may be found even premonocyte, premonoblast, nucleated rbc may also bee seen but in small number ✿ BC prep is well suitable for the study of morphology of plateltes and white cells in case oof thrombocytopenia, leucopenia ✿ Buffy coat prep are also used to detect the abnormal circulating elements such as cancer cells, lymphosarcoma cells, fragments of megakaryocytes ✿ Buffy coat films can be used for the detection of bacyeria fungi or parasite within neutrophil, monocytes or circulating macrophage.
- 40. THE BUFFY COATIS THE FRACTION OF AN ANTICOAGULATED BLOOD SAMPLE THAT CONTAINS MOST OF THE WHITE BLOOD CELLS AND PLATELETS
- 41. PROCEDURE ✿ Venous blood/anticoagulatedblood is taken in a wintrobes haematocrit tube/ Capillary tube and centrifuged in 300rpm for 15min. ✿ After centrifugation remove thr supernatant serum carefully by fine pipette and with same pipette deposit the platelet anf underlying leuvocute layer on to slides ✿ Emulsify buffy coat with patients plasma and then spread a film ✿ Allow to dry in air and then fix and stain asusual ✿ As an alternative to centrifugation the blood may be allowed to sediment with the helpof sedimentation enhancing agents like fibrinogen, methyl cellulose, Boyum's reagent(leucocye prep)
- 43. IMPORTANCE OF BUFFYCOAT PREPRATION ✿ Buffy coat prep has very much imp bcoz it include only platelet and wbc ✿ In normal case the thickness of buffy coat layer is 0.1mm. The variation in the thickness of buffy coat layer is the imp infication of variation in the wbc and platelets ✿ Buffy coat prep helps in the identification of cancer cells, lymphosarcoma cells, fragments of megakaryocytes and erythrophagocytic cells. ✿ It reflect the conditions of thrombopenia, thrombocytosis, leucocytosis and leucopenia, buffy coat layer is greenish in colour
- 44. ✿ In normalcase plasma is straw yellow colour, yellow green, orange colour of plasma indicates increased bilirubin ✿ Milky or cloudy plasma layer inficates nephrosis and lipaemia
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