SamplingSamplingAccurate laboratory diagnosis is based on a valid sampling proceAccurate laboratory diagnosis is based on a valid sampling processss

Valid sampling process composed ofValid sampling process composed of::

-- Selection of appropriate sampleSelection of appropriate sample

-- Proper collection and handling of different types of samplesProper collection and handling of different types of samples

-- Proper labeling of the sampleProper labeling of the sample

-- Proper preservation of the samplesProper preservation of the samples

-- Avoidance of spoilage of samplesAvoidance of spoilage of samples

Purpose of samplingPurpose of sampling::

For helping to establish a disease diagnosis.For helping to establish a disease diagnosis.

For health surveillance and certification.For health surveillance and certification.

For monitoring the response to treatment or vaccination. For monitoring the response to treatment or vaccination.

II-- Selection of appropriate samplesSelection of appropriate samples

-- Selection of representative animalsSelection of representative animals

-- Animal that correctly represent the disease conditionAnimal that correctly represent the disease condition

-- Animal in advanced stage of the diseaseAnimal in advanced stage of the disease

-- In herd problem, samples from several animals with different In herd problem, samples from several animals with different

stages of the diseasestages of the disease

-- Collect samples from one or two recently died animalsCollect samples from one or two recently died animals

-- Selection of representative samplesSelection of representative samples

-- collected samples must be related to the diseased condition.collected samples must be related to the diseased condition.

-- More than one type of samples could be used if applicable.More than one type of samples could be used if applicable.

-- Avoid sample contamination.Avoid sample contamination.

-- Use of appropriate container for sample collection (Clean and dUse of appropriate container for sample collection (Clean and dry).ry).

-- Use of appropriate preservative for shipping of samples.Use of appropriate preservative for shipping of samples.

IIII-- Proper handling of different types of samplesProper handling of different types of samples

Type of samplesType of samples

Blood, Milk, Urine, Feces, Ruminal juice, Saliva and Blood, Milk, Urine, Feces, Ruminal juice, Saliva and

sputum and throat swap, Pus and exudates, Semen, sputum and throat swap, Pus and exudates, Semen,

preputial wash, Vaginal discharge, Skin scraping and preputial wash, Vaginal discharge, Skin scraping and

hair, hair, ……………………

11-- Blood samples:Blood samples:

-- Venous blood is preferred for most hematological examinations.Venous blood is preferred for most hematological examinations.

-- They are collected aseptically in different forms using specialThey are collected aseptically in different forms using special venule, venule,

vacutainer tubes or plastic syringes.vacutainer tubes or plastic syringes.

Site of collection:Site of collection:HorseHorse Jugular vein.Jugular vein.DonkeyDonkey Jugular vein.Jugular vein.BuffaloesBuffaloes Jugular vein.Jugular vein.Sheep & Sheep & GootGoot Jugular vein.Jugular vein.DogDog Cephalic vein, lateral Cephalic vein, lateral saphenoussaphenous vein.vein.CatCat Ear vein, femoral vein.Ear vein, femoral vein.PigPig Ear vein, anterior vena cava.Ear vein, anterior vena cava.RabbitRabbit Marginal ear vein.Marginal ear vein.Rat and mouseRat and mouse Tail vein.Tail vein.

Precautions during collection of blood samples:Precautions during collection of blood samples:

-- The site of collection of blood must be clean, dry and sterile.The site of collection of blood must be clean, dry and sterile.

-- All the equipment used in collection must be clean, dry and steAll the equipment used in collection must be clean, dry and sterile.rile.

-- The blood sample must be collected on the wall of the tube to The blood sample must be collected on the wall of the tube to

avoid the hemolysis.avoid the hemolysis.

-- Gentle mixing of the blood sample with the anticoagulant.Gentle mixing of the blood sample with the anticoagulant.

-- Avoid contamination of the blood samples with hairs or dirties.Avoid contamination of the blood samples with hairs or dirties.

-- The blood sample must be transferred directly to the laboratoryThe blood sample must be transferred directly to the laboratory

under suitable conditions.under suitable conditions.

Errors occur during collection of blood samples:Errors occur during collection of blood samples:

-- Using wet needle or syringe.Using wet needle or syringe.

-- Taking too much time in collection of blood.Taking too much time in collection of blood.

-- Failure to mixing the blood sample with anticoagulant immediateFailure to mixing the blood sample with anticoagulant immediately after ly after

collection or filling vials to the top. not properly mixed with collection or filling vials to the top. not properly mixed with blood.blood.

-- Vigorous mixing of the blood sample.Vigorous mixing of the blood sample.

-- Excessive negative pressure when collecting sample with a syrinExcessive negative pressure when collecting sample with a syringe will ge will

rupture cells and collapse the vein.rupture cells and collapse the vein.

-- Failure to remove the needle from the syringe, when transferrinFailure to remove the needle from the syringe, when transferring blood g blood

from a syringe to a container.from a syringe to a container.

Types of blood samplesA) Whole blood. B) Serum.

C) Plasma. D) Blood smear.

E) Blood swab.

A)A)WHOLE BLOODWHOLE BLOOD

(Anti(Anti--coagulated blood)coagulated blood)

Blood mixed with anticoagulant usually dipotassium salt of EDTA Blood mixed with anticoagulant usually dipotassium salt of EDTA

and used for:and used for:

1.1. Hematological pictureHematological picture 2. Blood culture.2. Blood culture.

3. Blood smear3. Blood smear 4. Biochemical determination.4. Biochemical determination.

5. Blood transfusion.5. Blood transfusion.

BB-- SERUM and PLASMASERUM and PLASMA

-- For biochemical analysis and serological examinationFor biochemical analysis and serological examination

SerumSerum from coagulated blood (Whole blood without anticoagulant)from coagulated blood (Whole blood without anticoagulant)

PlasmaPlasma from nonfrom non--coagulated blood (whole blood with anticoagulant)coagulated blood (whole blood with anticoagulant)

STORAGE OF PLASMA OR SERUMSTORAGE OF PLASMA OR SERUM

-- Samples may be stored at 4 Samples may be stored at 4 °°C in a refrigerator for up to 4 days or in freezing C in a refrigerator for up to 4 days or in freezing

compartment for up to 1 week.compartment for up to 1 week.

-- If stored for longer periods, samples should be placed into If stored for longer periods, samples should be placed into

deep freeze at deep freeze at --20 20 °°C.C.

D) BLOOD SMEARD) BLOOD SMEARSignificance of the blood filmSignificance of the blood film

-- Identification of animal species.Identification of animal species.

-- Morphological classification of anemia.Morphological classification of anemia.

-- Diagnosis of bacterial diseases.Diagnosis of bacterial diseases.

-- Diagnosis of blood parasites.Diagnosis of blood parasites.

-- Deferential leucocytic counts (DLC).Deferential leucocytic counts (DLC).

E) Blood swabsE) Blood swabs-- For bacterial culture.For bacterial culture.

-- Blood swab is taken from heart blood of small animals.Blood swab is taken from heart blood of small animals.

-- In Anthrax, blood swab obtained from venous blood by tamponIn Anthrax, blood swab obtained from venous blood by tampon

Tests performed on blood samplesTests performed on blood samplesAA-- Hematological examination:Hematological examination:-- Total erythrocytic count.Total erythrocytic count. -- Total leucocytic count.Total leucocytic count.-- HematocritHematocrit (Packed cell volume).(Packed cell volume). -- Differential leucocytic count.Differential leucocytic count.-- Determination of blood hemoglobin.Determination of blood hemoglobin. -- Erythrocytic sedimentation rate.Erythrocytic sedimentation rate.-- Platelets count.Platelets count. -- Bleeding time.Bleeding time.-- Coagulation time.Coagulation time. -- Blood grouping.Blood grouping.

BB-- Biochemical examination (plasma or serum):Biochemical examination (plasma or serum):Serum is preferred since it is less likely to show hemolysis thSerum is preferred since it is less likely to show hemolysis than plasma and an plasma and contain no anticoagulants which draw water outside the cells.contain no anticoagulants which draw water outside the cells.

CC-- Bacteriological examination.Bacteriological examination.Collected blood is directly injected into the bottle containing Collected blood is directly injected into the bottle containing the culture media. the culture media.

DD-- Parasitological examination.Parasitological examination.Stained blood smear is commonly used.Stained blood smear is commonly used.Diagnosis of both intraDiagnosis of both intra--cellular and extracellular and extra--cellular blood parasites. cellular blood parasites.

EE-- Toxicological examination.Toxicological examination.

FF-- Serological examination:Serological examination:For estimation of the antibody level against certain diseaseFor estimation of the antibody level against certain disease-- producing producing organisms.organisms.

GG-- Blood transfusionBlood transfusion

22-- Milk samplesMilk samples

During collection of milk sample avoid external contamination byDuring collection of milk sample avoid external contamination by::

-- Thorough cleaning of udder and teat orifice.Thorough cleaning of udder and teat orifice.

-- Collection in clean and sterilized containers.Collection in clean and sterilized containers.

Separate sample collected from each quarter (diseased or normal)Separate sample collected from each quarter (diseased or normal)..

To avoid misdiagnosis, samples collected in the following order To avoid misdiagnosis, samples collected in the following order (LF, LH, RH, (LF, LH, RH, RF).RF).

Immediately following collection, milk sample kept in refrigeratImmediately following collection, milk sample kept in refrigerator till time or till time analysis.analysis.

Indication of milk sampleIndication of milk sample::

-- Examination of chemical and physical characteristics.Examination of chemical and physical characteristics.

-- Bacteriological examination for bacterial count and detection oBacteriological examination for bacterial count and detection of mastitis f mastitis causative causative

agent.agent.

-- Detection of Brucella antibodies (Detection of Brucella antibodies (AbortusAbortus Bang ring test Bang ring test –– ABR test)ABR test)

33-- Urine sampleUrine sample

Urine sample can be collected throughUrine sample can be collected through::-- Clean catch method after stimulation of animal urination.Clean catch method after stimulation of animal urination.

-- Catheterization for less contaminated samples.Catheterization for less contaminated samples.

-- Cystocentesis for aseptic sample.Cystocentesis for aseptic sample.

Indication of urine sample:Indication of urine sample:-- Routine urine analysis.Routine urine analysis.

-- Bacterial examination.Bacterial examination.

-- Parasitic examinationParasitic examination..

44-- Fecal sampleFecal sample

In large animalsIn large animals:: fresh sample collected directly from animal by back fresh sample collected directly from animal by back

racking using plastic bag.racking using plastic bag.

In small animalsIn small animals:: samples collected with:samples collected with:

-- Finger covered with gloves.Finger covered with gloves.

-- Sterile fecal spoon or fecal swap.Sterile fecal spoon or fecal swap.

-- Rectal enema with worm water.Rectal enema with worm water.

Collected fecal sample should be examined directly or refrigeratCollected fecal sample should be examined directly or refrigerated.ed.

Preservation include addition of 10% formalinPreservation include addition of 10% formalin

Indication of fecal sampleIndication of fecal sample::

-- Examination of internal parasite infestation.Examination of internal parasite infestation.

-- Evaluation of digestive system.Evaluation of digestive system.

-- Bacteriological exam. (isolation of Salmonella or smear for aciBacteriological exam. (isolation of Salmonella or smear for acid fast bacilli).d fast bacilli).

-- Chemical and toxicological examination.Chemical and toxicological examination.

55-- Skin scraping and hairSkin scraping and hair

Skin scraping for diagnosis of mangeSkin scraping for diagnosis of mange::

-- Scraping of the periphery of the lesion till oozes of blood usiScraping of the periphery of the lesion till oozes of blood using dry or ng dry or moist scalpel (mineral oil or glycerin). moist scalpel (mineral oil or glycerin).

-- Skin scraping collected in screw capped bottle, test tubes or PSkin scraping collected in screw capped bottle, test tubes or Petri dishes. etri dishes.

-- 10% NaOH added for maceration of tissues and clearance of paras10% NaOH added for maceration of tissues and clearance of parasite.ite.

Hair sample for diagnosis of Ring wormHair sample for diagnosis of Ring worm::

-- Pull a tuft of hair but not cut to obtain the root of the hair.Pull a tuft of hair but not cut to obtain the root of the hair.

-- Wrap in a paper or put in envelop or in clean test tube.Wrap in a paper or put in envelop or in clean test tube.

Ruminal juiceRuminal juice

Saliva and sputum and throat swapSaliva and sputum and throat swap

Pus and exudatesPus and exudates

SemenSemen

preputial washpreputial wash

Vaginal dischargeVaginal discharge

Other types of samples:Other types of samples:

IIIIII-- Proper labeling of the sampleProper labeling of the sample11-- Data of the animalData of the animal

-- Type, Sex, AgeType, Sex, Age

22-- Data of the ownerData of the owner

-- Name, Address, DateName, Address, Date

33-- Data of the physicianData of the physician

-- Name, locationName, location44-- Data of the samplesData of the samples

-- Type of sample.Type of sample.

-- Preservatives or other chemicals used with the samplePreservatives or other chemicals used with the sample

55-- Tentative clinical diagnosis and main clinical signsTentative clinical diagnosis and main clinical signs

66-- Desired examinationsDesired examinations

IVIV-- Proper preservation of samplesProper preservation of samples

Type of preservatives:Type of preservatives:-- Physical preservativesPhysical preservatives

-- Chemical preservativesChemical preservatives

Proper preservation is to keep samples till time of analysis in Proper preservation is to keep samples till time of analysis in a a

condition similar to that when obtained first time.condition similar to that when obtained first time.

AA-- Physical preservatives:Physical preservatives:

11-- Refrigeration:Refrigeration:

By keeping the specimen in the refrigerator at 4By keeping the specimen in the refrigerator at 4˚̊c.c.

Suitable for short time preservation (few hours)Suitable for short time preservation (few hours)

22-- Natural or dry ice:Natural or dry ice:

-- Natural iceNatural ice::

* can preserve specimens from 12 to 24 hours.* can preserve specimens from 12 to 24 hours.

* Specimen placed in water tight container and surrounded by ice* Specimen placed in water tight container and surrounded by ice..

-- Dry iceDry ice: (Solid Co: (Solid Co22))

* Dry ice can provide longer preservation.* Dry ice can provide longer preservation.

* Specimen placed in plastic bag or water tight container and dr* Specimen placed in plastic bag or water tight container and dry ice y ice wrapped in a paper and placed in the box.wrapped in a paper and placed in the box.

* Avoid direct contact with the specimen.* Avoid direct contact with the specimen.

* Dry ice is not to be used in air proof container to avoid expl* Dry ice is not to be used in air proof container to avoid explosion osion from the volatile gases and pressure.from the volatile gases and pressure.

33-- Freezing:Freezing:

-- Freezing provide the longest preservation time. Freezing provide the longest preservation time.

-- Suitable for preservation of specimens to be used for bacteriolSuitable for preservation of specimens to be used for bacteriological examination.ogical examination.

-- Used for preservation of serum and plasma samples for long timeUsed for preservation of serum and plasma samples for long time (deep freezing).(deep freezing).

-- Not to be used for parasitic examination of feces or hematologiNot to be used for parasitic examination of feces or hematological examination of cal examination of

whole blood.whole blood.

BB-- Chemical preservativesChemical preservatives

11-- Fixing solutionsFixing solutions::

•• 10% aqueous solution of formalin or 95% ethyl alcohol.10% aqueous solution of formalin or 95% ethyl alcohol.

•• 10 times volume of fixative should be added to the 10 times volume of fixative should be added to the spicmenspicmen..

•• Penetrates the tissues and results in Harding and preservation Penetrates the tissues and results in Harding and preservation for long time.for long time.

•• Suitable for histological examination of tissues.Suitable for histological examination of tissues.

•• Fixing solution for viral examination composed of normal salineFixing solution for viral examination composed of normal saline ( 0.85% ( 0.85% NaclNacl) )

containing 1% gelatin.containing 1% gelatin.

22-- Bactericidal solutionBactericidal solution::•• Chemicals used when keeping bacterial growth to the minimum is Chemicals used when keeping bacterial growth to the minimum is

desired.desired.•• Not to be used with specimens for bacteriological examination.Not to be used with specimens for bacteriological examination.•• This include: This include:

* formalin 10%: for fecal and urine samples.* formalin 10%: for fecal and urine samples.* Phenol 0.5%: for serum sample.* Phenol 0.5%: for serum sample.

-- Chemical substances added to blood samples to keep blood in liqChemical substances added to blood samples to keep blood in liquid form.uid form.

-- The best anticoagulant is the one which prevent coagulation witThe best anticoagulant is the one which prevent coagulation with least cellular h least cellular

damage.damage.

-- The choice of anticoagulant depends on the type of examination The choice of anticoagulant depends on the type of examination to be carried to be carried

out.out.

33-- AnticoagulantsAnticoagulants

The most important anticoagulants areThe most important anticoagulants are::

Ethylene Ethylene DiamineDiamine TetraTetra--acetic acid (EDTA).acetic acid (EDTA).

Heparin.Heparin.

Ammonium and potassium oxalate mixture.Ammonium and potassium oxalate mixture.

Sodium citrate.Sodium citrate.

Sodium fluoride and potassium oxalate mixture.Sodium fluoride and potassium oxalate mixture.

VV-- Avoidance of sample spoilageAvoidance of sample spoilage

CAUSES OF SPECIMEN SPOILAGECAUSES OF SPECIMEN SPOILAGE

-- Autolysis.Autolysis.

-- Hemolysis.Hemolysis.

-- Fragmentation.Fragmentation.

-- Drying (Desiccation).Drying (Desiccation).

-- Decomposition.Decomposition.

11-- AutolysisAutolysis-- It is the digestion of the sample by its own enzymes. It is the digestion of the sample by its own enzymes. -- It happens most often in samples of the digestive tract. It happens most often in samples of the digestive tract. -- It may occur in samples packed in borax or some other dry antisIt may occur in samples packed in borax or some other dry antiseptic powder.eptic powder.Autolysis is helped byAutolysis is helped by::-- High temperature and is directly related to worm climate.High temperature and is directly related to worm climate.-- Time between collection in the field and receipt at the laboratTime between collection in the field and receipt at the laboratory. ory.

22-- HemolysisHemolysis-- It is the breakdown of the cellular elements in the blood samplIt is the breakdown of the cellular elements in the blood samples.es.CausesCauses::-- Using wet needle or syringe.Using wet needle or syringe.-- Collection of the blood sample directly to the bottom of the tuCollection of the blood sample directly to the bottom of the tube.be.-- Vigorous mixing of the blood sample.Vigorous mixing of the blood sample.-- Excessive negative pressure when collecting sample with a syrinExcessive negative pressure when collecting sample with a syringe. ge. -- Failure to remove the needle from the syringe. Failure to remove the needle from the syringe. -- Bacterial contamination.Bacterial contamination. -- Chemical contamination.Chemical contamination.-- Extreme heat or cold.Extreme heat or cold.

33-- FragmentationFragmentation-- It is breaking the sample into small pieces.It is breaking the sample into small pieces.-- This results fromThis results from::

-- Forcing a specimen into a small bottle.Forcing a specimen into a small bottle.-- Cutting the specimen with dull knife or with scissors.Cutting the specimen with dull knife or with scissors.

44-- DryingDrying-- Drying occurs in certain types of samples such as blood, serum,Drying occurs in certain types of samples such as blood, serum, exudates or pus.exudates or pus.-- This results fromThis results from

-- Too small sample.Too small sample.-- Too large container.Too large container.

55-- DecompositionDecomposition-- Slight over growth by bacteria or mold can make material unfit Slight over growth by bacteria or mold can make material unfit for examination.for examination.-- Bacterial or fungal enzymes frequently digest tissues and destBacterial or fungal enzymes frequently digest tissues and destroy both roy both

structural and cellular organization.structural and cellular organization.-- This results fromThis results from::

-- Contamination with soil, Contamination with soil, faecesfaeces or intestinal contents.or intestinal contents.-- Long time in shipment.Long time in shipment.-- High temperature.High temperature.-- Bacterial contamination.Bacterial contamination.

Thank youThank you

1.1. Ethylene Ethylene DiamineDiamine TetraTetra--acetic acidacetic acid ((DisodiumDisodium or dipotassium salt of EDTA)or dipotassium salt of EDTA)

Mode of actionMode of action:: Precipitation of calcium ions.Precipitation of calcium ions.

AdvantagesAdvantages::

-- Excellent preserving power, recommended for routine blood examinExcellent preserving power, recommended for routine blood examination. ation.

-- DoesnDoesn’’t alter the erythrocyte size and allow excellent leucocytes stat alter the erythrocyte size and allow excellent leucocytes staining.ining.

-- Used in determination of creatinine, urea nitrogen, glucose, phUsed in determination of creatinine, urea nitrogen, glucose, phosphorus and uric acid.osphorus and uric acid.

DisadvantagesDisadvantages::

-- Higher concentration of salt withdraws water from red cells andHigher concentration of salt withdraws water from red cells and reduces PCV values.reduces PCV values.

-- Not suitable for determination of alkaline phosphatase (ALP) acNot suitable for determination of alkaline phosphatase (ALP) activity as it combine with Mg ions tivity as it combine with Mg ions

needed for activation of ALP.needed for activation of ALP.

-- Chloride estimation in EDTA blood always higher than that with Chloride estimation in EDTA blood always higher than that with other anticoagulants.other anticoagulants.

Amount requiredAmount required::

1mg / 1 ml blood. (for 5 ml blood, add 0.5 ml of 1 % solution or1mg / 1 ml blood. (for 5 ml blood, add 0.5 ml of 1 % solution or 1 drop of 10% solution to each tube 1 drop of 10% solution to each tube

and allow the water to evaporate ofand allow the water to evaporate off).f).

2. 2. HEPARINHEPARIN-- It is a natural anticoagulant, found abundantly in the liver frIt is a natural anticoagulant, found abundantly in the liver from which its name is derived.om which its name is derived.

Mode of actionMode of action: :

Prevents blood coagulation by interfering with conversion of proPrevents blood coagulation by interfering with conversion of prothrombin into thrombin.thrombin into thrombin.

AdvantagesAdvantages::

-- Does not alter the erythrocyte volume (suitable for PCV).Does not alter the erythrocyte volume (suitable for PCV).

-- Least effect on erythrocyte Least effect on erythrocyte haemolysishaemolysis.. -- Recommended for blood of cats.Recommended for blood of cats.

Disadvantages:Disadvantages:

-- More expensive.More expensive. -- Unsuitable for smears for its poor stain affinity. Unsuitable for smears for its poor stain affinity.

-- Its action stopped after 8 hr.Its action stopped after 8 hr. -- Cause the cells to stain bluish with wrights stain.Cause the cells to stain bluish with wrights stain.

Amount requiredAmount required::

-- For 5 ml blood, add 0.1 ml of 0.75 % solution and evaporate to For 5 ml blood, add 0.1 ml of 0.75 % solution and evaporate to dryness at room temperature. dryness at room temperature.

(Can be used in liquid form as (Can be used in liquid form as HeparineHeparine injection to coat the inside of the syringe).injection to coat the inside of the syringe).

3. AMMONIUM AND POTASSIUM OXALATE MIXTURE3. AMMONIUM AND POTASSIUM OXALATE MIXTURE(HELLER AND PAUL MIXTURE)(HELLER AND PAUL MIXTURE)

Mode of actionMode of action:: Binding ionized calcium.Binding ionized calcium.

AdvantagesAdvantages::-- It is cheaper than EDTA.It is cheaper than EDTA. -- Easy to prepare and use.Easy to prepare and use.-- Little cellular distortion occurs if the sample is examined witLittle cellular distortion occurs if the sample is examined within the 1st hour of collection.hin the 1st hour of collection.-- Cause very little hemolysis or changes in the volume of Cause very little hemolysis or changes in the volume of RBCsRBCs..

DisadvantagesDisadvantages::-- It doesnIt doesn’’t prevent clumping of platelets.t prevent clumping of platelets. -- It is poisonous.It is poisonous.-- It canIt can’’t be used in estimation of the potassium, ALP or urea.t be used in estimation of the potassium, ALP or urea.

Amount requiredAmount required::-- Ammonium oxalate 1.2 gm.Ammonium oxalate 1.2 gm.-- Potassium oxalate 0.8 gm.Potassium oxalate 0.8 gm.-- D.W. 100 ml.D.W. 100 ml.

1ml of the solution in a tube, then dry at 60 1ml of the solution in a tube, then dry at 60 °°C. This is sufficient for 10 ml blood.C. This is sufficient for 10 ml blood.or, 2 mg / ml blood.or, 2 mg / ml blood.

4. SODIUM CITRATE4. SODIUM CITRATE

Mode of actionMode of action:: Binding ionized calcium.Binding ionized calcium.

AdvantagesAdvantages::

-- It is employed in blood transfusion as citrate is metabolized aIt is employed in blood transfusion as citrate is metabolized and excreted rapidly.nd excreted rapidly.

DisadvantagesDisadvantages::

-- Interfere with many biochemical tests.Interfere with many biochemical tests. -- Prevent the clotting for only few hr.Prevent the clotting for only few hr.

-- Increased concentration may cause shrinkage of cells.Increased concentration may cause shrinkage of cells.

Amount requiredAmount required::

For blood transfusion: For blood transfusion: TriTri--sodium citrate 1.32 gm.sodium citrate 1.32 gm.

Citric acid 0.48 gm.Citric acid 0.48 gm.

Dextrose 1.4 gm.Dextrose 1.4 gm.

D. W. D. W. 100 ml.100 ml.

-- The strength of the stock solution is 3.8 %.The strength of the stock solution is 3.8 %.

(9 volumes of blood + 1 volume of sodium citrate solution and mi(9 volumes of blood + 1 volume of sodium citrate solution and mixed immediately)xed immediately)

-- Sodium citrate is also widely used in estimation of the ESR.Sodium citrate is also widely used in estimation of the ESR.

( 4 volumes of venous blood + 1 volume of sodium citrate solutio( 4 volumes of venous blood + 1 volume of sodium citrate solution) n)

5. 5. SODIUM FLUORIDE AND POTASSIUM OXALATE MIXTURESODIUM FLUORIDE AND POTASSIUM OXALATE MIXTUREMode of actionMode of action:: Binding ionized calcium.Binding ionized calcium.

AdvantagesAdvantages::

-- Most suitable for determination of glucose level, since it is eMost suitable for determination of glucose level, since it is effective in inhibiting the glycolytic ffective in inhibiting the glycolytic

enzyme which breakdown glucose in blood.enzyme which breakdown glucose in blood.

DisadvantagesDisadvantages::

-- It is poisonous.It is poisonous.

-- Unsuitable for determination the level of ALP or urea determinaUnsuitable for determination the level of ALP or urea determination.tion.

Amount requiredAmount required::-- It is better to use the mixture than sodium fluoride alone as iIt is better to use the mixture than sodium fluoride alone as it increase the t increase the anticoagulant effect.anticoagulant effect.

-- The mixture consists ofThe mixture consists of:: 4 parts Sodium Fluoride 4 parts Sodium Fluoride 5 parts Potassium Oxalate.5 parts Potassium Oxalate.

-- For 5 ml. blood, add 0.5 ml of 2.25 % solution of the mixture aFor 5 ml. blood, add 0.5 ml of 2.25 % solution of the mixture and evaporate off nd evaporate off the water.the water.