Yacutainer Its blood collection tube which is a sterile glass or plastic tube with a closure that is evacuated to create a vacuum inside the tube facilitating the draw of a predetermined volume. Most commonly used to draw the blood samples directly from the vein. Vacutainer tubes may contains additives, mostly the anticoagulants, designed to stabilize and preserve the blood specimen prior to analytical testing. These tubes are available with or without a safety-engineered closure (the rubber stopper inside the plastic cap), with a variety of labeling options and closure colors as well as a range of draw volumes. 4 Cer ByHere is the list of Color codes of the tube, the Additives present in them and their common uses — COLORCopES = ADDITIVE Purple / : EDTA (Ethylene diamine tetra-acetic axe Lavender - Light blue Tri-Sodium Citrate (Blood ; Anticoagulant ratio - 9:1) Light green Lithium Heparin (COMMONLY USED FOR w CBC, BLOOD TYPING (Rh Factor & ABO Screening), Cross match, Hb, Red cell Indices, ESR by Wintrobe's method etc. Prothrombin time (PT), Activated Partial Thromboplastin Time (APTT), Fibrinogen thrombin time and other blood Coagulation tests. Basic Metabolic Panel (BMP), ‘Comprehensive metabolic Panel (CMP) and other plasma determination testsHere is the list of Color codes of the tube, the Additives present in them and their common | uses— COLOR CODES ADDITIVE COMMONLY USED FOR Red 7) (No additive/Plain tube BMP, CMP, Lipid Profile, a Serology tests, Therapeutic AS drug monitoring, blood bank procedures, etc. Dark Sodium Heparin Arterial blood gas analysis, Fema a alpha-TNF, Lymphocyte O o Immunotherapy etc. P|Here is the list of Color codes of the tube, the Additives present in them and their common uses— COLOR CODES Gray ADDITIVE Sodium Fluoride Tri-Sodium Citrate (Blood: Anticoagulant ratio- 4:1) COMMONLY USED FOR Blood Sugar testing, Toxicology tests etc. ESR by Westergren methodRBC pipette Its a calibrated pipette used to draw a very small sample of blood from a finger prick, dilute it with specialized fluid (Hayem’s fluid) and apply a droplet of it to a special calibrated hemocytometer (Neubauer’s chamber) to count the RBC. th Pipette has a capillary stem with uniform bore and bulb for dilution and mixing and is calibrated to deliver 20ul or 0.02m! of blood. Bulb is attached to red rubber mouth piece. f A bulb contains a red bead which facilitates the mixing of blood with é diluting fluid. Freely mobile bead indicates dry pipette. 4:2 Marking on pipette are 0.5, 1, 101. “<WBC pipette Calibrated pipette used to draw a very small sample of blood from a finger prick, dilute it with specialized fluid and apply a droplet of it to a special calibrated hemocytometer (Neubauer’s chamber) to count the WBC. Pipette has a capillary stem with uniform bore and bulb for dilution and mixing and is calibrated to deliver 20ul or 0.02ml of blood. Bulb is attached to rubber mouth piece which is white in colour. ‘A bulb contains a white bead which facilitates the mixing of blood with diluting fluid. Freely mobile bead indicates dry pipette. Marking on pipette are 0.5, 1, 11. 4 Z o‘A thick glass having central platform divided into positions with the help of H shaped groove or trench. + On both the sides of the lateral groove there are raised ridges of a height of 0.1mm (1/10 mm) from the central platform. + When a cover slip is placed on the ridges, a space of 0.1mm height is created below the cover slip on the central platform. * Counting chamber (Grid) is made up of ruled area of 3x3mm size on each central platform. + Each central area is further divided by triple lines into 9 squares of equal size (Imm? each). + Four comer squares are further divided into 16 squares of equal size & are used to do total leucocyte count (TLC). + The volume of each big square is 0.1mm? (1x0.1 mm). + Central big square is divided into 25 (medium size) squares. + Area of each medium size square is 1/25 mm (1/5 x 1/5). < * Volume of each square is 1/250 [Link] (1/25 x 1/10). + Further, these medium size square are divided into 16 small squares of equal size. + Area of each small square is 1/20 mm? ( 1/5 x 1/4). + Area of each square is 1/400 mm? (1/20 x 1/20) and volume is 1/4000 mm? (1/400 x 1/10).‘A thick glass having central platform divided into positions with the help of H shaped groove or trench. * On both the sides of the lateral groove there are raised ridges of a height of 0.1mm (1/10 mm) from the central platform. * When a cover slip is placed on the ridges, a space of 0.1mm height is created below the cover slip on the central platform. * Counting chamber (Grid) is made up of ruled area of 3x3mm size on each central platform. + Each central area is further divided by triple lines into 9 squares of equal size (Imm? each). + Four comer squares are further divided into 16 squares of equal size & are used to do total leucocyte count (TLC). + The volume of each big square is 0.1mm? (1x0.1 mm). * Central big square is divided into 25 (medium size) squares. + Area of each medium size square is 1/25 mm? (1/5 x 1/5). < * Volume of each square is 1/250 [Link] (1/25 x 1/10). + Further, these medium size square are divided into 16 small squares of equal size. + Area of each small square is 1/20 mm? ( 1/5 x 1/4). + Area of each square is 1/400 mm? (1/20 x 1/20) and volume is 1/4000 mm? (1/400 x 1/10).Total number of smallest squares in big central square are 400 (25 x 16). RBCs are counted in 5 medium size square R1, R2, R3, R4 i.e. four comers and one central.Sal Hemoglobinomete: Ithas following apparatus: 1) Comparator box- This has brown colored rods along with empty area to fit hemoglobinometer tube. 2) Hemoglobinometer Tube- It may be round or square glass tube. Round hemoglobinometer tube has marking in from 2 to 22 on one side and 10-140 on other side. The result should be expressed in gmv/dl as no single value of Hb is considered 100% since it varies with age, sex & geographical location. 3) Hemoglobinometer pipette- This is glass pipette having one marking of 20ul along with rubber teat. 4) N/10 HCL. This contains N/10 HCl. 5) Stirrer- This is a glass rod for mixing. 6) Dropper - to put distilled water in hemoglobinometer tube. Reagents ~ 1) N/10 HCL- This contains N/10 HCL, 2) Distilled water Principle ~ Hb is converted to brown color acid hematin the color of which is compared with brown colored standard rods in comparator box. 4Procedure — 1) Fill Sabli’s hemoglobinometer tube with N/10 HCI up to mark 2gms. 2) Take 20u! of blood in Sabli’s Hemoglobinometer pipette. Wipe extra blood present on tip of Sahli’s Hemoglobinometer pipette with absorbent paper or gauze and deliver it to Sahli’s hemoglobinometer tube. 3) Stir with stirrer and wait for 10 minutes. 4) Add Distilled water drop by drop & stir till colour matches with the comparator. 5) Take the reading at lower meniscus. Results ~ The hemoglobin value of the given blood sample is ........gm/dl Inference — The hemoglobin value is normal/ lower! higher. Advantages ~ 1) Simple method & can be done bed side 4: 2) Less expensiveDisadvantages- 1) Since it is manual comparison of color change, there can be visual error. 2) Carboxy, met. & sulthemoglobin cannot be converted to acid hematin, So obtained value is lower than the actual value. 3) Glass comparator can fade over the years. 4) Colour of acid hematin also fades away quickly and this can give low Hb value. 5) Source of light (daylight or artificial) will influence the visual comparison of colors. ‘Normal Values — Hemoglobin — Adult Male- 13.6 - 17.2 gmidl, Adult Female~ 12,0 - 15.0 gmv/dl, Adult pregnant females~ 11.0 - 14.0 gm/dl, Atbirth- 13.6 - 19.6 gm/dl Crier} 4:Glass slide: o Thin flat piece of glass used to hold the object for examination under microscope. © Size of slide is 75 x 25 mm; thickness about Imm. © These are often used with a cover slip, which is smaller and a thinner sheet of glass, and is placed over the tissue mounted on the glass slide. © Glass slide allows to manipulate the tissue and other specimen in a small space. Uses: © For making blood smears. © For mounting of histopathological tissue. © For mounting the cytopathological samples. feCoverslip: These are the thinner sheet of glass used to cover the mounted specimen on the Slide. Uses: © To keeps the solid specimen pressed flat and liquid specimen of even thickness. © To secure the slide for longer time . © It prevent the contamination of the specimen mounted on the slide; by airborne particle or others. © Decreases the evaporation rate of wet preparations.Wintrobe tubes: © The Wintrobe tube is a glass tube closed at one end. The tube is 110mm long and has an internal bore diameter of 2.5mm. © The tube is graduated on both sides: from 0 to 10cm on one side and 10 to Oem on the other. © Anticoagulants: Either of the following 2 anticoagulants can be used. 1. Ethylene diamine tetraacetic acid (EDTA) solid crystals 1-2 me/ml. 2. Double oxalate (solid) 2-3 mg/ml (ammonium oxalate and sodium or potassium oxalate in the ratio of 3:2.Wintrobe’s method Draw 2ml of blood into the anticoagulant. Fill Wintrobe tube upto mark 0 with anticoagulated blood with the help of a Pasteur pipette having a long stem (15 em) so as to fill the tube free of air bubbles 3. Place the tube vertically in a stand and note the ESR after one hour. pe Normal values: Males 0-7 mm 1* hour Females 0-15 mm 1" hour Advantages i, Itis simple method and requires small amount of blood. There is no dilution with anticoagulant. . Packed cell volume (PCV) can also be done by the same tube. iv. Filling of tube with Pasteur pipette eliminates chance of any infection due to handling of blood. Disadvantages: i, Because of short column and choice of anticoagulant, it is not as sensitive index of diseases as Westergren’s. ii, Addition of more anticoagulant can lower ESR. iii, ESR of more than 100mm cannot be measured. 4:© Westergren’s pipette is a straight pipette 30 cm long open at both ends with internal bore diameter of 2.5mm and is calibrated from 0-200mm from top to bottom. © Anticoagulant Trisodium citrate as 3.8g/dl liquid { anticoagulant is used. It is used in the concentration of 1 four parts of blood are added to one part of 7 anticoagulant. Westergren’s pipette \ Uses: ESR Estimation Procedure: Principle- When well mixed anticoagulated blood is placed in a vertical tube, the erythrocytes tend to fall towards the bottom of the tube/pipette till they form a packed column in the lower part of the tube in a given time. 4: ' i 4Westergren’s Method: 1. The patient is advised to come in the moming fasting (as heavy protein diet affects concentration of plasmaproteins). 2. Take 1.6ml of patient's blood and mix it with 0.4m of citrate anticoagulant already put in a tube. The test should be done within two hours of taking blood. 3. Fill the pipette upto mark 0 with citrated blood with the help of rubber teat by vacuum filling and fix it in a rack vertically away from sunlight or vibrations. 4. Let it stand for one hour after which reading is taken at the upper meniscus of the RBCs. Normal values: Males 3-Smm 1" hour; Females 4-7mm 1 hour Advantages: lt is a more sensitive method. 2iIt is easy to fill and clean the Westergren’s pipette. Disadvantages: [Link] more amount of blood. 2. Dilution of blood in anticoagulant affects ESR. 4 3 Filling of blood by mouth pipetting should be strictly discouraged. ‘ABO Anti Sera- Principle- Human red blood cell possessing A or B antigen will agglutinate in the presence of antibody directed towards the antigen. Agglutination of the RBCs with Anti-A, Anti-B or Anti-AB reagents is a positive test results and indicates the presence of the corresponding antigen Anti sera A_and B: © Itis a ready to use solution having monoclonal antibodies of immunoglobulin class IgM, prepared from hybridoma cell line, created by fusing mouse antibody producing B lymphocytes with mouse myeloma cells. Each hybridoma cell line produces homogenous antibodies of only one immunoglobulin class, which is identical in their chemical structure and immunological activity. Titre— More than or equal to 256 (1:256) Each vial is of 10ml. Preservative used- 0.1% Sodium Azide. Grouping is concentration dependent. Store- At 2-8 degree. DO NOT FREEZE. Shelf life of the regent is as per the expiry date mentioned on the regent vial label. 4: . eo ceceAnti sera D-(Rho)- # Itis a ready to use solution having monoclonal antibodies of immunoglobulin class IgM and IgG, prepared from hybridoma cell line, created by fusing mouse antibody producing B lymphocytes with mouse myeloma cells or from human B cell line through EBV transformation. «Each hybridoma cell line produces homogenous antibodies of only one immunoglobulin class, which is identical in their chemical structure and immunological activity. «These antibodies are monoclonal antibodies of same specificity but having the capability of recognising different epitopes of the human RBC s anitgen D. Titre — More than or equal to 256 (1:256) Each vial is of 10m. Preservative used- 0.1% Sodium Azide Grouping is not concentration dependent. ‘Use:To detect the blood group of the patient. Methods of the blood grouping: Slide method, Tube Method & Gel method <LITMUS PAPER:- oe Itis used to test whether a solution is acidic or basic. Blue litmus paper turns red under acidic conditions and red litmus paper turns blue under basic or alkaline conditions with the color change occurring over the ph range 4.5-8.3 at 25 degree Celsius(77 degree F). ‘Neutral litmus paper is purple Litmus is a dye made from naturally occurring substance including liches (fungi)- the most commonly used of which is the species Roccella tinctoria. Hold one end of a piece of red litmus paper, dip it into the sample and remove it. There is no need to leave in the sample for any length of time because any color change is immediate. Repeat the step with a piece of blue litmus paper In the absence of litmus paper, reagent phenolphthalein can be used.SALAH BONE MARROW ASPIRATION NEEDLE :- SITE FOR BONE MARROW ASPIRATION- - Posterior iliac crest, - Anterior superior iliac crest, -Tibia (only in infants younger than 1 years), = Sternum (last resort in morbid obese and older than 12 years) INDICATIONS- ‘+ Unexplained anemia, abnormal red cell indices, cytopenia or cytoses Abnormal peripheral blood smear morphology suggestive of bone marrow pathology Diagnosis, staging and follow up of malignant hematological disorders (eg. acute and chronic leukemias, myelodysplastic syndrome, chronic myeloproliferative disorders, lymphomas, plasma cell myeloma, amyloidosis, mastocytosis) Suspected bone marrow metastasis Unexplained focal bony lesion on radiological imaging Unexplained organomegaly or presence of mass lesion inaccessible for biopsy Microbiological culture for investigation of pyrexia of unknown origin or specific infection. Eg- Miliary Tuberculosis, Leishmaniasis, Malaria Evaluation of iron stores Investigation of lipid /glycogen storage disorders Exclusion of hematological disease in potential allogenic stem cell transplant disorders.LUMBAR PUNCTURE NEEDLE: USES. ‘© Collect CSF sample (spinal tap) for biochemical, microbiological and cytological analysis ‘© Spinal anaesthesia ‘© Therapeutic lumbar puncture (to relieve increased ICT) SITE OF INSERTION. between L3 and L4 in adults DIAGNOSTIC INDICATIONS. © Meningitis © Pyrexia of unknown origin ‘© Subarachnoid hemorshage ‘© -Myelography-introduction of radiological dyes, ‘THERAPEUTIC INDIACTIONS- Spinal Anaesthesia, Chemotherapy of leukemia & ‘Administration of intracranial antibodies CONTRAINDICATIONS. Increased ICT, Deformity of spine, Local infection & Bleeding diathesisJAMSHIDI BONE MARROW BIOPSY NEEDLE:- (TREPHINE BIOPSY NEEDLE) ‘© Used for bone marrow core biopsy for study of marrow ‘components by paraffin embedding technique Site- posterior iliac crest Staining- hematoxylin and eosin, reticulin on tissue sections Time-within 1-7 days Morphology- better marrow architectural pattern but cell morphology is not distinct since tissue sections are examined and not smears eee * INDICATIONS. Myelosiderosis, aplastic anemia, in case with “dry tap’ on aspiration.‘VIM SILVERMAN’S NEEDLE:- Used for tissue core biopsy from liver. It has three parts :- Cannula, Stellate / Trocar, Prong / Fork / bifid needle. Indications:- In evaluation of jaundice Liver cirrhosis Storage disorder - Glycogen storage disorder, Hemochromatosis, Wilson’s disease Granulomatous lesions like TB and sarcoidosis Infections- Viral (CMV, Herpes) Parasitic (anaerobic liver abscess) To diagnose benign and malignant neoplasmContraindications:. Bifid needle Bleeding diathesis Hydatid cyst Severe ascites Trocar /Stilette Sites:- 9% and 10" intercostals space in mid axillary area, CanulaURINOMETE! ‘Used for estimation of specific gravity of urine There are three parts of urinometer. They are as illustrated in the Figure J. The float: the air containing part AL Weight: the lower end of urinometer, HL. Stem. Hias calibrations with numbers marked to measure the specific gravity The weight is a bulb filled with a ball bearings embedded in are solid, probably a glass of some sort The glass stem extends in a tube of urine and where the meniscus of the urine reduces displays the specific gravity of urine Typically used in medical diagnostic labs. Itis based on the principle of BUOYANCY. Because of increased density of urine compared to that of water, the urinometer will float higher in urine than in water. The weighted float will displace the volume of the urine which is equal to its weight. The ‘weight of the weighted float is designed in such a way that the BEEERREEEERThe steps are as follows: i. Allow urine to reach room temperature 4 ii, Fill the glass cylinder with urine up to 2/3rds. At least 15 ml of urine is required iii, Make sure that there are no bubbles/foam in the glass cylinder. This may lead to difficult to take the readings of the meniscus iv. Gently insert the urinometer into the container. Make sure the urinometer do not touch the bottom or the sides of the container and allow it to float freely. v. One may even have to gently spin the urinometer so that it is in the center of the cylinder vi. Read the calibrated graduation at the lowest level of urinary meniscus. vii. Note that urinometer is calibrated at 15°C so the temperature correction has to be done to measure the correct specific gravity.URISTICKS. Reagent coated strips for rapid testing of ingredients in urine It determines the pathological changes in a patients urine in standard urinalysis A standard urine test strips may comprises upto 10 different chemical pads or reagents which react (change color) when immersed in, and then removed from a urine sample. Uristicks provide tests for glucose, bilirubin, ketones, specific gravity, blood, pH, protein, urobilinogen, nitrite and leukocytes in urine.ESBACHS ALBUMINOMETER_ + It is used for quantitative estimation of albumin in urine. + The graduation indicate grams of protein per litre of urine. Urine is placed in tube up to U level and then Esbachs reagent (mixture of picric and citric acid in water) is poured in upto R mark. From the base upto the marking ‘U’, the tube is graduated from 0-12gm. Principle: The albumin in the urine combine with the picric and citric acid in the Esbachs reagent and forms precipitate which is measured quantitatively. Precautions: Make sure that the urine is acidic. R-Reagent U- Urine 0-12 Marking from base upwards