Uncategorized

Uncategorized

Cavities of the body

/

  The human body cavity is a fluid-filled space inside the body that holds and protects internal organs. The body maintains its internal organization by membrane,sheaths,and other structure that separate the compartments. The two main divisions are the dorsal cavity (posterior) and ventral  cavity  (anterior).These cavities protect the lungs,heart,stomach,and intestines for example can expand and contract without distorting other tissues or disrupting the activity of nearby organs. The posterior (dorsal) cavity has two main subdivisions:- 1.cranial cavity :- The cranial cavity is the hollow space inside the skull  and protects the brain.The cranial cavity contains the brain,protective membranes called meninges. cerebrospinal fluid (CSF) blood vessels supplying the brain.its called as houses of the brain.its formed by the bones of the skull especially the cranium protected by skull and cerebrospinal fluid. Functions of the Cranial Cavity:- Brain Protection: The solid, bony walls (calvarium) and fluid-filled membranes act as a shock-absorbing enclosure for the brain, cerebellum, and brainstem. Structural Support: The base of the skull forms the floor, consisting of the anterior, middle, and posterior cranial fossae, which provide a stable, shaped foundation where different parts of the brain rest. Muscular Attachment: The exterior surface of the cavity provides anchoring points for muscles, including the temporalis muscle, which is vital for jaw movement.  Environmental Stability: It creates a stable, insulated environment necessary for delicate brain functions, including the regulation of temperature, hormonal signals via the pituitary gland, and blood flow. vertibral cavity:-The spinal cavity (vertebral cavity) encloses the spinal cord .its a long, narrow space inside the vertebral column (backbone) Protected by the vertebral column and cerebrospinal fluid.It extends from the base of the skull to the lower back.the vertibral cavity main function is protect the spinal cord from injury and provides a pathway for spinal nerves.its supports communication between the brain and the rest of the body through the nervous system. Functions of the Vertebral Cavity:- Protection: The primary function is to serve as a strong bony encasement for the delicate spinal cord, protecting it from injury. Support & Structure: It forms a rigid central axis for the trunk, providing a safe housing for nerve tissue while supporting the body’s upright posture. Path for Nerves: The cavity enables nerve roots to pass out through the intervertebral foramina, facilitating connection to the peripheral nervous system. Space and Cushioning: It provides a contained space for the spinal cord, which is padded and held in place by cerebrospinal fluid and the meninges. Accommodation of Structure: The cavity allows for flexibility and movement of the vertebral column while maintaining the structural integrity of the spinal cord.   The anterior (ventral) cavity has two main subdivisions:- 1.Thoracic cavity:-The thoracic cavity is situated between the neck and diaphragm in the upper part of the trunk.its boundaries are formed by the thoracic cage and supporting muscles.contain vital organs involved in respiration and circulation.(like-Trachea,2 bronchi,2 lungs,Heart, aorta, superior and inferior venacavae,blood vessels, oesophagus. The thoracic cavity is divided into three main compartments:- A. Right Pleural Cavity:- It contain the right lungs lined by  pleura. B. Left Pleural Cavity :-  It contain the left lungs also lined by pleura. C. Mediastinum (central compartment):-The Mediastinum is  space between the  lungs including the structures found there,sach as- heart oesophagus, and blood vessels Function of Thoracic cavity:- Protection: The rib cage, sternum, and thoracic vertebrae form a rigid, bony structure that shields vital organs such as the heart and lungs from injury. Respiration Facilitation: The cavity is designed for breathing (pulmonary ventilation). The diaphragm and intercostal muscles work to change the volume of the thoracic cavity, creating pressure changes that move air into and out of the lungs. Support & Movement: The cavity provides a stable anchor for the arms and protects the superior thoracic aperture (neck area), while remaining flexible enough for the torso to bend and twist. Organ Housing: It contains key cardiovascular components (heart, great vessels), respiratory components (lungs, trachea, bronchi), and the esophagus 2. Abdominopelvic  cavities:-The abdominopelvic cavity is a large body cavity located below the diaphragm and above the pelvic floor is commonly divided into two parts: Abdominal cavity – the upper portion Pelvic cavity – the lower portion A.Abdominal cavity–The abdominal cavity is the upper part of the abdominopelvic cavity. The abdominal cavity is a large,fluid-lined body space that houses and protects the vital organs like-digestive,urinary,and reproductive organs. its the large hollow space in the body located between the chest(Thoracic cavity) and the pelvis. By convention,the abdominal cavity is divided into the nine regions:- 1.Epigastric region 2.Umbilical region                                                                                                                                                                                                                                                    3.Hypogastric region                                                                                                                                                                                                                                                          4. Left hypochondriac region         

Uncategorized

METACHROMATIC STAINING

/

INTRODUCTION:- There are certain basic dyes belonging to aniline group that will differentiate particular tissue components by giving them a different color to that of original dye. The phenomenon is known as metachromasia. Some of the common metachromatic dyes are:  *Methylene blue, Methyl violent                                                                                                                                *Thionin, Crystal violet ,                                                                                                                                            *Toluidine blue Metachromasia :- takes place when certain negatively charged groups on the tissue react with cationic dyes. On polymerization the original colour of the dye changes to another colour (eg mast cell stain pink with toluidine blue). Thionin and toluidine blue dyes are commonly used for quick staining of frozen selection using their metachromatic property to stain nucleus and cytoplasm differently. Metachromasia is enhanced when intermolecular distances are reduced.  Factors which enhance metachromasia are 1. Increasing concentration of dye.  2. Decreasing temperature.  3. pH  4. Water a polar solvent, contributes to the efficiency of van der Waal’s forces by which the molecules are held together. In tissues, where there is a high concentration of anions e.g. in sulphatedmucopolysaccharides, the cationic dye molecules may be held in such closeproximity to one another that van der Waal’s forces can exert their influence andcause the dye to polymerize. Consequently the colour changes from blue to red. Tissue components often demonstrated by metachromatic stains: *Amyloid material, Mast cell granules.                                                                                                                           *Mucin Cartilage Amyloid Stain -Various stains are used to demonstrate amyloid  CRYSTAL VIOLET STAIN FOR AMYLOID:-  Aim: To demonstrate amyloid in tissue sections. Principle: Amyloid (a glycoprotein) exhibits metachromasia in tissue section when stained with crystal violet and other cationic dyes. Control: ositive control ReagentsCrystal violet solutionStock solutionCrystal violet                                         14 gm95% alcohol                                           100 mlWorking solutionStock solution                                        10 mlDistilled water                                        300 mlConcentrated hydrochloric acid         1 ml  Procedure:-  Deparaffinize and bring the sections to water. Put working crystal violet solution for 1 to 2 minutes and check undermicroscope. Rinse in tap water. Mount in water or in water soluble media. Put on the coverslip seal the edges with nail polish (Do not let it dry.) Result:- Amyloid                             purple violetOther tissues                    blue CONGO-RED STAIN FOR AMYLOID Aim: To demontrate amyloid in tissues. Principle: Diazo dye attaches itself to amyloid fibrils. The union is affected byH bonds between the OH groups of amyloid and amino side groups of the dye.Congo red dye forms non-polar hydrogen bonds with amyloid. The greenbirefringence of congo red stained amyloid by polarized light is considereddiagnostic of amyloid. Control: Known positive tissue Reagents:-Congo red solution Congo red                                         1.0gmDistilled water                                  100ml Saturated solution of Lithium Carbonate Procedure:- *Bring section to water. *Pour congo red solution for 20 minutes. *Pour off the solution and cover the slide with lithium carbonate for 1.5 minutes to differentiate. *Wash with water. *Counter-stain with hematoxyline for 5 minutes. *Differentiate with 1% acid alcohol. *Wash in running tap water. *Dehydrate, clear in xylene and mount in DPX. Result :- Amyloid                                                              bright red which gives apple green birefringence in polarized light.Nuclei                                                                 blueOther structures                                               unstained to yellow Notes :- 1. Sections must be cut at 8 to 10 microns for birefringence  2. Solution must be filtered through glass wool, not paper filters for birefringence to occur  3. Tissue fixed in solutions other than formalin may display false positive birefringence

Histopathology, Uncategorized

Decalcification in histology

/

INTRODUCTION:- The presence of calcium salts in tissues makes them hard. This causes damage to the knife, difficulty in cutting tissue. Calcium is normally present in bones and teeth. Calcium may also be present in normal tissues in pathological conditions like necrotic tissue in tuberculosis. OBJECTIVES:- After reading this lesson, you will be able to:  *describe decalcification *explain different methods of decalcification *describe the chemical and physical tests to estimate the remaining calcium. DECALCIFICATION:- Aim – To remove calcium salts from the tissues and make them amenable for sectioning.  Preparation of tissues – The calcified hard tissues should be first cut into small pieces (2 to 6mm) with a thin blade, hacksaw or sharp knife in order to minimize the tearing of the surrounding tissues. This process is followed by fixation in buffered formalin or any other desired fixative. After fixation tissues must be thoroughly washed and excess fixative should be removed before the specimen is subjected to decalcification. DIFFERENT METHODS OF DECALCIFICATION:- 1. Acid decalcification 2. Ion exchange resin 3. Electrical ionization 4. Chelating methods 5. Surface decalcification Decalcification process should satisfy the following conditions *Complete removal of calcium salts *Minimal distortion of cell morphology *No interference during staining Decalcification is a straightforward process but to be successful it requires: *A careful preliminary assessment of the specimen *Thorough fixation *Preparation of slices of reasonable thickness for fixation and processing *The choice of a suitable decalcifier with adequate volume, changed regularly *A careful determination of the endpoint *Thorough processing using a suitable schedule. Methods of Decalcification :- The tissue is cut into small pieces of 3 to 5 mm size. This helps in faster decalcification. The tissue is then suspended in decalcifying medium with waxed thread. The covering of wax on thread prevents from the action of acid on thread. The volume of the decalcifying solution should be 50 to 100 times of the volume of tissue. The decalcification should be checked at the regular interval. Acid Decalcification – This is the most commonly used method. Various acid solutions may be used alone or in combination with a neutralizer. The neutralizer helps in preventing the swelling of the cells. Following are the usually used decalcifying solutions- 1. Aqueous Nitric Acid:- Nitric acid              –         5ml Distilled water       –        100 ml If tissue is left for long time in the solution, the tissue may be damaged. Yellow colour of nitric acid should be removed with urea. But this solution gives good nuclear staining and also rapid action. 2. Nitric Acid Formaldehyde Nitric acid                 – 10 ml Formaline                 – 5-10 ml                                                                                                                        Distilled water upto 100 ml Advantages   * Rapid action *Good nuclear staining *Washing with water is not required *Formalin protects the tissues from maceration 3. Formic Acid Solution: Formic acid                   – 5  ml Distilled water              – 90 ml Formalin                       –  5 ml In this solution the decalcification is slow. If concentration of formic acid is increased the process is fast but tissue damage is more: 4. Trichloroacitic Acid – This is used for small biopsies. The process ofdecalcification is slow hence cannot be used for dense bone or big bony pieces. Formal saline (10%)                 –      95 ml Tricloroacitic acid                    –      5 gm Ion Exchange method – In these ammonium salts of sulfonated polystyrene resin is used. The salt is layered on the bottom of the container and formic acid containing fluid is filled. The decalcifying fluid should not contain mineral acid. X-rays can only determine complete decalcification. The advantages of this method are:- *Faster decalcification *Well preserved tissue structures *Longer use of resin Electrolytic Method – Formic acid or HCl are used as electrolytic medium. The calcium ions move towards the cathode. Rapid decalcification is achieved but heat produced may damage the cytological details. Chelating Agents:- Organic chelating agents absorb metallic ions. EDTA can bind calcium forming a non-ionized soluble complex. It works best for cancerous bone. This is best method for decalcification of bone marrow biopsies as it preserves cytological details best. The glycogen of marrow is preserved. EDTA Solution :- EDTA                                    –                 5.5 gm Formaline                          –                  100 ml Distilled water-                 –                   900 ml Surface Decalcification – The surface layer of paraffin blocks are inverted in5% HCl for one hour. About top 30 micron is decalcified. It should be washed thoroughly before cutting. Factors affecting rate of Decalcification:- 1. Concentration of decalcifying solution-Increased concentration of the decalcifying agent fastens the reaction. 2. Temperature-The rate of decalcification increases with rise of temperature. 3. Density of bone-Harder bone takes longer time to decalcify. 4. Thickness of the tissue-Small tissue pieces decalcify earlier. 5. Agitation-Agitation increases the rate of decalcification. METHODS OF DETERMINING OPTIMUM DECALCIFICATION OR ENDPOINT:-  Specimens should NOT be crowded together and should NOT contact the bottom of container in order to provide complete decalcification. Over decalcification can also permanently damage specimen. The following procedure help determine the correct end-point of decalcification. End-Point of Decalcification: X-ray (the most

Uncategorized

RAITMAN- FRANKEL CALORIMETRIC (SGOT)

/

SERUM SGOT SUMMERY:-     Serum Glutamate Oxaloacetate Transaminase (SGOT), also called as Aspartate aminotransferase (AST), belongs to the transferase class of enzymes. This enzyme shows high levels of activity in the heart, liver, skeletal muscles and kidneys. Since its level seems to be increasing enormously following Myocardial Infarction (MI), it can be used as supporting evidence in the diagnosis of MI (Especially 20-36 hrs after MI). Elevated levels are also seen in Viral / Toxic Hepatitis, Hepatic and Cardiac Necrosis, Muscular Dystrophy and Pulmonary Embolism. AIM:- Determination of Serum Glutamate Pyruvate Transaminase (SGPT or ALT) by Reifman & Frankel’s method  PRINCIPLE:- Alanine aminotransferase (GPT) catalyzes the transfer of the amino group from alanine to oxoglutarate with the formation of glutamate and pyruvate.   L-Aspartate + 2-Oxogluatarate                      GPT            Oxaloacetate +L-Glutamate        Oxaloacetate + 2,4 DNPH                                              Brownish red colored complex SGOT (AST) catalyses the transfer of amino group from Aspartic acid to 2- Oxoglutarate to form Oxaloacetate and L-Glutamate. The Oxaloacetate thus formed reacts with 2,4 Dinitrophenyl Hydrazine (2,4 DNPH) to form a corresponding Hydrazone, a brownish red colored complex in an alkaline medium.The color intensity is directly proportional to the SGOT concentration in the serum and is measured photometerically at 505 nm (490- 546).    REQUIREMENTS:-     ·        Four test-tube, ·        Colorimeter, ·        SGOT Reagent Kit, ·        Incubator, ·        Cuvette, ·        Pipette etc. ·        Fresh Serum PROCEDURE:-   1.      Pipette into test tubes labeled as Blank, Calibrator, Control, Test and proceed as per given below       Reagent Blank Calibrator Control Test Substrate Reagent 0.25 ml 0.25 ml 0.25 ml 0.25 ml Deionized Water 50µl       Serum Sample       50µl Calibrator (Conc. 170 U/L)   50µl      1.      Mix and incubate at 37ºC for 60 minutes. Color Reagent 0.25 ml 0.25 ml 0.25 ml 0.25 ml Serum Sample (Same Serum Sample Which isused above)     0.05 ml   1.      Mix and incubate at 37ºC for 20 minutes. Alkaline Reagent (Prediluted) 1.5 ml 1.5 ml 1.5 ml 1.5 ml 1.      Read absorbance of all the tubes against distilled water at 505 nm(490-546).   Calculation:-:- SGOT (AST) activity in U/L =                 Abs of Test – Abs of Control  x Conc of Calibrator (Conc :160 U/L)                              Abs of Calibrator– Abs of Blank Then,    If Absorption of,        Test                       =             0.35                                      Control                 =             0.30                                      Calibrator            =             0.36                                     Blank                     =             0.05 Then,  SGOT (AST) activity in U/L =            Abs of Test – Abs of Control      xConc of Calibrator (Conc :160 U/L)                                                       Abs of Calibrator– Abs of Blank Then,       SGOT (AST) activity in U/L =            0.35 –0.30  x 160 U                                                                 0.36 – 0.5          SGOT(AST) activity in U/L =          25.8   Result (U/L)                        =             25.8 NORMAL VALUE:-        =          0-35 U/L                    CLINICAL SIGNFINANCE The group of enzymes called transaminase exist in tissues of many organs. Necrotic activity in these organs causes a release of measured. Since heart tissue is rich in AST increased serum levels appear in patients after myocardial infraction, as well as in patients with muscle disease. Muscular dystrophy and dermatomyositis . The liver is especially rich in ALT, being this enzyme measurement used primarily as a test for infectious and toxic hepatitis, although high levels of both ALT and AST may also be found in cases of liver cell damage and acute pancreatitis, suggesting that the obstruction of billary tree by the adematous pancreas and the presence of associate hepatic disease may contribute to elevated AST levels these patients. Slight or moderate elevations of AST and ALT activities may be observed after intake of alcohol and after administration of various drugs, such as salicylates, opiates, and ampicilin

Microbiology, Uncategorized

Methyl Red Test FOR ENTEROBACTERIA

/

METHYL RED TEST :- This test is performed to differentiate Enterobacteria. v  Principle:- Some Enterobacteria when cultured in buffered glucose peptone water, ferment glucose to produce sufficient acidity, which gives a red color with methyl red indicator (pH range: 4.4-6.2. Color change: red to yellow). v Procedure:- 1.      Inoculate a colony of the test organism into 0.5 ml of sterile glucose phosphate broth. 2.      Incubate overnight at 35-37°C. 3.      Add a drop of methyl red indicator and observe the color. v Observations:- 1        Bright red color                  :                Positive test 2        Yellow/orange color         :                 Negative test v Quality control:- Use following microorganisms to confirm the reliability of reagents: Positive control      :               E. coli Negative control    :               Klebsiella aerogenes

Microbiology, Uncategorized

NITRATE REDUCTION TEST

/

NITRATE REDUCTION TEST:- This test helps to differentiate bacteria that produce the enzyme nitrate reductase from the bacteria that do not produce the enzyme. This test is also helpful in differentiating Mycobacterium species. v      Principle:-       The test organisms are incubated in a broth containing nitrate. The nitrate reductase producing organisms reduce nitrate to nitrite, which is tested by adding sulfanilic acid reagent and a-naphthylamine. The formation of pink red compound indicates positive reaction.    v Requirements:-        1.      Nitrate broth         2.      Sulfanilic acid reagent                 a) Glacial acetic acid                        : 5.7 ml  b) Distilled water                              :  14.3 ml c) Sulfanilic acid                                :  0.16 g  3. alfa-naphthylamine  reagent a) Glacial acetic acid                        :  5.7 ml b) Distilled water                               : 14.3 ml c) Sulphonic acid                               :0.16 g v     Procedure:- 1.      Incubate the nitrate broth (sterile) with test organism. 2.      Incubate at 37°C for four hours 3.      Add one drop of sulfanilic acid reagent. 4.      Add one drop of a naphthylamine reagent. 5.      Mix well and observe the reaction   v Observations:-        Red color                            : Positive test         No red color                       :No reduction of nitrate   v Additional information:-      When nitrate is not detected it is necessary to test whether the organism has reduced the nitrate beyond nitrite to nitrogen gas or ammonia. Zinc dust (knife point), small amount is added, which will convert any nitrate to nitrite. In that case the observation is as follows:       Red color             : Negative test       No red color        : Positive test v    Quality control:- Use the following microorganisms to confirm the reliability of reagents:- Positive control        : E.coli Negative contro l      : Acinetobacter sp.

Microbiology, Uncategorized

INDOLE TEST FOR BACTERIA IDENTIFICATION

/

INDOLE TEST :- This test is important in the identification of enterobacteria such as E.coli, P. vulgaris, P. rettgeri, etc. v Principle:- The test organism is cultured in a medium containing tryptophan. The organisms break down tryptophan and indole is released. It is detected by the action of Kovac’s or Ehrlich reagent (formation of red colored compound. This test can also be performed by culturing the organism in tryptone water or peptone water containing tryptophan. Indole production is detected as described above). v   Requirements:- 1.      Motility indole urea (MIU) medium 2.      Kovac’s reagent strips (or Kovac’s reagent) Kovac’s Reagent v  Formula and preparation:- 1. p-dimethylaminobenzaldehyde            :        2.0 g 2. Isoamyl alcohol                                       :       30 ml 3. Concentrated hydrochloric acid           :       10 ml Dissolve ingredients 1 and 2 in 10 ml of concentrated hydrochloric acid and store in a brown bottle. v Procedure:- 1.      Inoculate MIU medium with test organism colonies. 2.   Incubate at 37°C by placing Kovac’s reagent strip in the neck of the MIU tube and 3 .       Look for reddening of the lower part of the test strip (or formation of red color of the reaction mixture). v Results:-  1. Reddening of strip                  : Positive test 2  No red color                              : Negative test v Quality control:- Use following microorganisms to confirm the reliability of reagents Positive control: Escherichia coli Negative control: Klebsiella aerogenes

Microbiology, Uncategorized

UREASE TEST FOR BACTERIA

/

Proteus strains are strong urease producers. Salmonellae and Shigellae do not produce urease. This test helps in differentiating enterobacteria. v    Principle:-  The test organisms are cultured in MIU medium (or in Christensen’s urea broth). If the strain produces urease, it acts on urea, and ammonium carbonate is formed with the release of ammonia. The medium becomes alkaline and the color of the medium changes to red pink. v  Requirements:- 1.      Motility Indole Urea (MIU) medium. 2.       Test tubes (15 x 125 mm) v  Procedure:- 1.      Inoculate MIU medium with acolony of the test organism. 2.      Incubate at 35°C overnight. 3.      Examine the medium by looking for a red pink color. v    Observations:- 1.     Red-pink medium       :  Positive test 2.     No red pink color        :  Negative test v  Quality control:-    Use the following microorganisms to confirm the reliability of reagents:-    Positive control     :           Klebsiella aerogenes    Negative control    :         E. coli

Microbiology, Uncategorized

BILE SOLUBILITY TEST

/

1.    Bile Solubility Test:- This test helps to differentiate S. pneumoniae from iridans streptococci.     v Principle:-        The test organisms are emulsified in normal saline. It gives a turbid suspension. The bile salt sodium deoxycholate is then added. S. pneumoniae organisms are soluble in bile salts and it is indicated by the clearing of turbidity. Viridans streptococci are insoluble in bile salts and it is indicated by persistence of the turbidity.    v Requirements:-        1.      10 g/dl sodium deoxycholate in 0.85 g/dl, sodium chloride.        2.  Normal saline.  3.   Test tubes (15 x 125 mm). v Procedure:- 1.      Pipette 2 ml of normal saline in a test tube. 2.      Suspend several colonies in the saline. 3.      Divide the suspension between two tubes. 4.      To the tube labeled as ‘T’, add 2 drops of sodium deoxycholate reagent, and to the tube labeled as ‘C’ add 2 drops of sterile distilled water. Leave both the tubes for 10-15 minutes. 5.      Observe the tubes. v Observation:- 1.      Clearing of turbidity: Probably S. pneumoniae. 2.      No clearing of turbidity: The organism is probably not S. pneumoniae.     v Quality control:-                Use the following microorganisms to confirm the reliability of reagents: Positive control: Streptococcus pneumonia                Negative control: Enterococcus fecalis  

Biochemistry, Uncategorized

SERUM DIRECT AND INDIRECT BILIRUBIN

/

SERUM DIRECT BILIRUBIN:-   Aim: – Determination of Serum Direct Bilirubin by DMSO method & Malloy and Evelyn method Summary:- Bilirubin is a breakdown product of hemoglobin, insoluble in water. It is transported from the spleen to the liver and excreted into bile. Hyperbilirubinemia results from the increase of bilirubin concentrations in plasma. PRINCIPLE:-   Bilirubin is converted to colored azobilirubin by diazotized sulfanilic acid and measured photometrically. Of the two fractions presents in serum, bilirubin-glucuromide and free bilirubin loosely bound to albumin, only the former reacts directly in aqueous solution (bilirubin direct), while free bilirubin requires solubilization with dimethylsulphoxide (DMSO) to react (bilirubin indirect). In the determination of indirect bilirubin, the direct is also determined; the results correspond to total bilirubin. The intensity of the color formed is proportional to the bilirubin concentration in the sample.  REQUIREMENTS:-   *Two test-tube, *Colorimeter, Bilirubin *Direct Reagents,  *Incubator, *Cuvette, *Pipette etc. *Fresh Serum. PROCEDURE-   1.      Adjust the instrument to zero with distilled water. 2.   Pipette into a cuvette or test tube: Contents Blank Test Direct Bilirubin Reagent (R2) 1.5 ml 1.5 ml Working Reagent (R3) – 50µl Sample /Calibrator 50µl 50µl 1.      Mix and incubate for exactly 5 minutes at room temperature.        2.      Read the absorbance (A). at 546 nm wavelength. Calculation:-  With Factor: Direct Bilirubin (mg/dl) =              (A) Sample – (A) Sample Blank x Factor Theoretical Factor Direct bilirubin = 14 Direct Bilirubin (mg/dl)  =             (A) Sample – (A) Sample Blank x 14 After Testing, If, (A) Sample = 0.21 (A)   Sample Blank = 0.16                                                                  =             0.05 X 14                                                                  =             0.7 mg/dl    D. Bilirubin in mg/dl to µmol/L   =             mg/dL x 17.1 =          0.7 X 17.1 =          11.97 µmol/L Normal Value:-    Direct Bilirubin (mg/dl)           =         <0.3 mg/dl Clinical Significance:-  Causes of hyperbilirubinemia:- Direct bilirubin: Hepatic cholestasis, genetic errors, hepatocellular damage. Indirect bilirubin:- Indirect Bilirubin (mg/dl)              =             Total Bilirubin – Direct Bilirubin =             1.146 – 0.7 =             0.446 mg/dl Or                      =             0.446 X 17.1 µmol/L =             7.62 µmol/L Normal Value:-    Indirect Bilirubin (mg/dl)        =         0.3  – 0.9 mg/dl

Scroll to Top