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INSTRUMENTS pathology mbbs 2nd professional.pptx
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- 3. LANCET • Double edgedblade or needle • Sharp pointed end with fixed depth • Disposable USE: • Make finger prick Collect capillary blood • Skin prick to test for allergies
- 4. NEEDLES & SYRINGES Uses •Blood collection • FNAC • Cytology- Sample fluid collection • Skin test for drug sensitivity • Medication
- 5. VACUTAINER NEEDLE &HOLDER
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- 9. • Instruments – Sahli’shemoglobinometer- standard brown glass blocks mounted on a comparator – Sahli’s hemoglobinometer tube- Graduated tube 2-22 G %,10-140% – Sahli’s pipette- Special pipette to measure out 20 microlitre of blood • Reagents – N/10 HCl – Distilled water
- 10. Sahli’s method /Acid hematin method Principle • Hb + N/10 HCl Acid hematin (brown colour) • Brown colour compared with standard brown glass reference blocks • Intensity of brown colour α amount of acid hematin α amount of Hb in the blood sample
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- 12. AMBER COLOURED BOTTLE •Protect light sensitive chemicals • N/10 HCl
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- 14. 1. RBC PIPETTE Uses •RBC count • Platelet count • WBC count when total count is very high
- 15. FEATURES • Markings –0.5,1 & 101 • Red bead • Blood is drawn upto 0.5 mark & diluent upto 101 mark • Dilution – 1:200 • Diluting fluid – Hayem’s fluid • Mercuric chloride (0.25g)- Fixes, preserves and prevents the growth of bacteria • Sodium chloride (0.5 g)- iso – osmolarity • Sodium sulfate (2.5 g)- Prevent clumping of RBC • Distilled water – 100 ml
- 16. 2. WBC PIPETTE •Uses . • Manual WBC count • Absolute eosinophil count • Sperm count • Any fluid – Synovial fluid count ,CSF
- 17. FEATURES … • Whiteglass bead in the bulb • Markings – 0.5 , 1 & 11 • Blood is drawn upto 0.5 mark & diluent upto 11 mark. • Dilution 1: 20 • Diluting fluid – Turk’s fluid • Glacial acetic acid (2ml)- Lyses RBCs • Gentian violet(1% sol,1 drp)- Stains WBC nuclei • Distilled water – 100 ml
- 18. RBC PIPETTE WBCPIPETTE Red bead White BEAD Larger bulb size Bulb size smaller Lumen size Smaller Lumen size larger Markings – 0.5,1 &101 Markings -0.5, 1 & 11 Dilution 1:200 Dilution 1:20
- 20. Improved NEUBAUER countingchamber Other Hemocytometer • Old Neubauer chamber • Fuchs- Rosenthal counting chamber • Thoma chamber • Burker chamber • Louis-Charles Malassez device
- 22. • Entire ruledarea – square of 3 mm side • 9 large squares – 1mm sides • Central square – subdivided into 400 smaller squares , 0.050mm (1/20) side • Each 5th line double line • Smallest squares – 25 groups of 16 squares each • Depth of squares on placing cover slip – 0.1 mm
- 23. 16 large squaresin central area 25 large squares in central area Triple lines are widely spaced Triple lines are closely spaced Separating lines are dull Separating lines are vivid and clear
- 25. THICK COVERSLIP PROPERTIES • Smoothsurface • Even thickness- 0.3mm, 0.4mm, 0.5mm • Fluid depth of 0.1mm • Sizes- 16x22mm, 22x23mm
- 26. CAPILLARY TUBES •Uses • Todetermine whole blood coagulation time •Procedure – Capillary blood from finger prick is collected into glass capillary tubes – Small sections of tube are broken off at timed intervals – Time when blood strings out between broken surface is taken as clotting time.
- 27. PACKED CELL VOLUME(PCV) OR HAEMATOCRIT • PCV is defined as ratio of volume of RBCs to that of whole blood and is expressed as percentage • Methods for Estimation of PCV 1. Macro method (Wintrobe’s method) 2. Micro haematocrit method (capillary tube 75mmx1mm) 3. Electronic method NORMAL: 40-50% males 38-45 %- females 36-42%- pregnant ladies RULE OF 3 and 9 Hb (gm/dl) x 3 = PCV RBC (million/cmm) x 9 = PCV
- 28. Macro (Wintrobe’s) Method Procedure •Fill the Wintrobe tube upto mark 10 with well mixed anticoagulated blood (EDTA) by Pasteur’s pipette free of air bubbles. • Centrifuge the tube at 2000-2300 g for 30 minutes. • After centrifugation layers are noted in the wintrobe tube • i. Uppermost layer of plasma. • ii. Thin white layer of platelets. • iii. Greyish-pink layer of leucocytes. • iv. Lowermost is the layer of RBCs. • Note the lowermost height of column of packed RBC layer and express it as percentage. Advantages of Macro Method • i. PCV and ESR can be measured simultaneously. • ii. Buffy coat can be prepared for other tests. • iii. By seeing the colour of plasma we can know about some of the pathological conditions • Jaundice it is yellow, • Haemolysis it is pink, • Hyperlipidaemia it is milky
- 29. PASTEUR PIPETTE • Glassdropper with tapered end fitted with a rubber bulb • Named after Louis Pasteur USE: • Fill Wintrobe’s tube with blood • Transfer liquid or chemicals
- 30. WINTROBE ‘s tube •Thick walled glass tube • 11 cm long , 2.5 mm internal diameter • Flat inner base • Marked at 1 mm interval from both sides, 0-10 and 10-0 • Holds 1 ml of blood • Venous blood is used • Anticoagulants- EDTA or heparin or wintrobe’s mixture
- 31. • Primarily forPacked Cell Volume • Erythrocyte Sedimentation Rate • Buffy coat smear preparation for- • LE cell phenomenon, • Abnormal cells – Aleukemic leukaemia • Malarial parasite Uses of wintrobe tube-
- 32. MICROHAEMATOCRIT METHOD • Capillarytube • 75mm length • 1mm internal diameter • Plain or heparinised • Tube sealed with plastic sealant, modelling clay or heat • Centrifuged at 12000 g for 5 mins
- 34. ERYTHROCYTE SEDIMENTATION RATE •It is defined as rate of sedimentation of RBC in 1 hour STAGES of ESR • Phase of rouleaux formation: Initial period of 10 minutes, • Phase of settling: In the next 40 minutes, settling of RBCs occurs at a constant rate. „ • Phase of packing: Last 10 minutes sedimentation slows and packing of the RBCs to the bottom occurs • FACTORS INCREASING ESR • Anemia (less negative charge – low zeta potential) • Macrocytosis • Elevated fibrinogen • High temperature • Tilted tube • FACTORS DECREASING ESR • Polycythemia • Microcytes sickle cells spherocytes • Low fibrinogen • Vibration
- 36. • Anticoagulant –3.8%Tri Sodium citrate • 4ml blood & 1 ml anticoagulant – 4:1 ratio • Iit is measured as length of the plasma column in mm when a vertical column of blood is kept undisturbed for one hour Methods of ESR • 1. Westergren’s method • 2. Wintrobe’s method • 3. Micro ESR method- capillary tube used • 4. Automated methods NORMAL VALUES • Males 0-15 mm 1st hour • Females 0-20 mm 1st hour
- 37. WESTERGREN’S TUBE • Pipettewith both ends open • Long tube- 300mm length • Markings on lower 2/3 from 0 to 200, from top to bottom • Internal diameter 2.55mm • Tubes are placed in stands in which series of rubber discs are attached
- 38. Procedure • The patientis advised to come in the morning fasting (as heavy protein diet affects concentraton of plasma proteins). • Take 1.6 ml of patient’s blood and mix it with 0.4 ml of citrate anticoagulant already put in a tube. • Fill the pipette upto mark O with citrated blood with the help of rubber teat by vacuum filling and fix it in a rack vertically away from sun light or vibrations • Let it stand for one hour after which reading is taken at the upper meniscus of the RBCs. ADVANTAGES • i. It is a more sensitive method. • ii. It is easy to fill and clean the Westergren’s pipette
- 39. Esrite ESR kit •Contents – Borosilicate glass tube- 25 cm, uncalibrated – Graduated aluminum screen – Polystyrene stands
- 40. ANTISERA FOR ABOAND RH BLOOD GROUPING • BLUE VIAL- Antisera-A • YELLOW VIAL- Antisera-B • RED VIAL – Antisera- AB • GREY/COLOURLESS VIAL- Anti-D USE: ABO Blood grouping Rh grouping
- 41. GLASS SLIDE • Measures7.5 x 2.5 x 0.1 cm Use: • Peripheral blood smear • Blood grouping • Reticulocyte count • Cytology smear • Urine microscopy • Histopathology slide
- 42. TEST TUBES Uses • Bloodgrouping- Tube method • Cytology- Fluid collection and centrifugation • Urine examination • MICROBIOLOGY culture
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- 45. Anticoagulant and PreservativeSolution : • CPD( Citrate-phosphate-dextrose) – 21days (shelf life) • CPD-Adenine(CPDA-1) – 35days • SAGM( Saline-adenine-glucose-mannitol) – 42days
- 47. BONE MARROW ASPIRATIONNEEDLE Salah bone marrow aspiration needle • Stout needle-stainless steel • 7-8 cm length • Stilette & guard
- 48. BM TREPHINE BIOPSYNEEDLE JAMSHIDI TREPHINE BIOPSY NEEDLE SITES: • Sternum • Iliac crest • Anterior and posterior iliac spines • Upper end of tibia • Spinous process of vertebra
- 49. Indications Contraindications • Unexplainedanaemias, granulocytopenia, thrombocytopenia,pancytopenia • Megaloblastic anaemia • Aplastic anaemia • Leukemias • MDS • Infections- enteric fever • Bone tumours n myeloma • Staging of lymphoma • Disseminated TB • Bleeding disorders • Local infections
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- 51. URINOMETER • Identification – Weightedcylinder-floats in urine – Stem & bulb – Markings 1.000 – 1.060 with divisions of 0.002 • Uses – Determine specific gravity of urine
- 53. • Procedure – 24hrsample, fill 3/4th of cylinder with urine – Place urinometer without touching sides – Take reading at eye level,noting the lower meniscus Normal specific gravity varies from 1.015 – 1.025 In severe renal diseases specific gravity become fixed at 1.010
- 54. CORRECTION For temperature-for every 3 degree rise- add 0.001, if fall- subtract 0.001 For protein, subtract 0.003 for every 1gm/dl For glucose, subtract 0.004 for every 1gm/dl For dilution multiply by the dilution factor
- 55. PIPETTE USES: • Blood sample-blood grouping • Chemicals • Urine chemical examination GRADUATED GLASS PIPETTE WITH RUBBER TEAT DISPOSABLE PLASTIC PIPETTE
- 56. Esbach’s Albuminometer • Quantitativeestimation of proteins in urine • Urine taken upto U mark • Esbach’s reagent upto Rmark • Stopper & keep for 24 hrs • Reading from the level of precipitation divided by 10= % of total proteins
- 58. • Reagent • Esbach’sreagent – picric acid citric acid Dist. water • Acetic acid • Uses – 24hr urine protein estimation • Identification – Markings U & R – Graduated in grams of dried albumin per 1000 ml
- 59. Lumbar puncture needle •10-12cm long with stillete • 22-24 gauge needle used frequently • L3 – L4 level Diagnostic purpose • Meningitis • Subarachnoid haemorrhage • Demyelinating diseases • Meningeal involvement by leukemia • Contrast medium introduction Therapeutic purposes • Giving antibiotics, anti cancer drugs • Giving anaesthesia
- 60. Absolute contraindications 🞭Infected skinover the needle entry site 🞭 Presence of unequal pressures between the supratentorial and infratentorial compartments Relative containdications 🞭Increased intracranial pressure (ICP) 🞭 Coagulopathy 🞭 Brain abscess
- 61. AYRE’S SPATULA • Woodenspatula used to collect Pap smear • U shape/concave on one end for cervical smear • Hormonal status • Premalignant & malignant lesions • Flat/convex on other end for • Vaginal wall scrapping • Buccal smear (barr body) • Specimen obtained by rotating the spatula over the ectocervix
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- 63. PERFORATED TISSUE PROCESSINGCASSETTE PLASTIC/METAL
- 64. Leuckharts ‘L’ moulds USE: Tissue embedding and blocking Each block contains a tissue piece carrying a identification label.
- 65. MICROTOME BLADE • Disposablemetal blade used in microtomes for sectioning • Usually 80mm long x 14 mm high x 0.35mm thick
- 66. BLOCK HOLDER • Usedto hold paraffin blocks for sectioning in microtome
- 67. LIVER BIOPSY NEEDLE •Aspiration needles – attached to a syringe- Menghini • Cutting needles- Vim silverman, Trucut needles Indications of liver biopsy • Metabolic liver diseases • Evaluation of abnormal liver enzymes • Acute liver failure of unknown cause • Hepatitis C and Hep B prognosis and therapeutics • Pyrexia of unknown origin • Nonalcoholic fatty liver disease • Focal lesions
- 68. Contraindications • Absolute • Uncooperativepatients • Bleeding tendencies pT ≥ 4 secs or INR > 1.5 or platelet count < 60,000 • Echinococcal disease suspect • Hemangioma or other vascular tumors • Relative • Ascites • Obesity • Infection in right pleural cavity and/or below right diaphragm
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