The document discusses the components and methods of routine urine analysis, emphasizing the importance of proper specimen collection, preservation, and testing procedures. It details the physical, chemical, and microscopic examination of urine, including tests for various constituents such as proteins, glucose, ketones, and bile salts. Clinical significance of abnormal findings in urine analysis is also highlighted, linking them to various medical conditions.

1. ROUTINE URINE ANALYSIS
PRESENTER – DR. S. SUNDARESAN
ASSISTANT PROFESSOR
TAGORE MEDICAL COLLEGE AND HOSPITAL

2. COMPONENTS OF BASIC ROUTINE
ANALYSIS
SPECIMEN
EVALUATION or
Adequacy of specimen
GROSS/ PHYSICAL
EXAMATION
CHEMICAL
EXAMINATION
SEDIMENT
EXAMINATION

3. SPECIMEN EVALUATION
1. PROPER LABELLING
2. PROPER SPECIMEN FOR REQUESTED EXAMINATION
3. PROPER PRESERVATIVE ( TOLUENE, BORIC ACID,THYMOL , FORMALIN, HYDROCHROLIC ACID)
4. VISIBLE SIGNS OF CONTAMINATION
5. TRANSPORTATION DELAYS CAUSING DETERIORATION

4. ADEQUACY OF SPECIMEN
The specimen should be properly collected in a clean container which should be
properly labelled with name of the patient, age and sex, identity number with date and
time of collection.
It should not show signs of contamination.
Specimen Collection
For routine examination a clean glass tube is used; for bacteriologic examination a
sterilized tube or bottle is required.
A mid-stream sample is preferable i.e. first part of urine is discarded and mid-stream
sample is collected
For 24 hours sample, collection of urine is started in the morning at 8 AM and
subsequent samples are collected till next day 8 AM

5. Methods of Preservation of Urine
i. Refrigeration at 4°C
ii. Toluene: Toluene is used 1 ml per 50 ml of urine. It acts by forming a
surface layer and it preserves the chemical constituents of urine.
iii. Formalin: 6-8 drops of 40% formalin per 100 ml of urine is used. It
preserves RBCs and pus cells. However, its use has the disadvantage that it
gives false-positive test for sugar.
iv. Thymol: Thymol is a good preservative; 1% solution of thymol is used. Its
use has the disadvantage that it gives false-positive test for proteins.
v. Acids: Hydrochloric acid, sulfuric acid and boric acid can also be used as
preservative.

6. ◦ URINE ANALYSIS
PHYSICAL CHEMICAL MICROSCOPIC
1.VOLUME 1. URINE PH 1.CELLS
2.COLOR 2. PROTEINS 2.CASTS
3.APPEARENCE 3. GLUCOSE 3.CRYSTALS
4.ODOR 4. KETONES 4. ABNORMAL CELLS
5.SPECIFIC GRAVITY 5. BLOOD, HAEMOGLOBIN & OTHER FORMED
HAEMOSIDERIN AND MYOGLOBIN ELEMENTS
6. BILIRUBIN
7. UROBILINOGEN

7. VOLUME
Average value- 700 -2000ml/day , night time not exceeding 400 ml
INCREASE DECREASE
POLYURIA = >2000ml/24 HRS OLIGURIA = <500ML/24 HRS
NOCTURIA= >500ML with specific gravity ANURIA= <100ML/24 HRS
<1.018
early renal failure. Less intake of water, dehydration, renal ischaemia.
Renal stones, tumours.
PATHOLOGICAL- PATHOLOGICAL-
1)Defective hormone regulation 1)PRE-RENAL
2)Defective salt water absorbtion 2)POST RENAL
3)Osmotic diuresis 3)RENAL PARENCHYMAL DISEASE

8. COLOUR :
- it is largely due to urochrome, urobilin and uroerythrin.
- Normal – pale yellow or amber.
PALE
YELLOW
STRAW COLOR
BROWN COLOR AMBER COLOR
DARK RED

9. Colour
Normally urine is clear, pale or straw-coloured due to pigment urochrome.
i) Colourless in diabetes mellitus, diabetes insipidus, excess intake of water. Dilution due to Excessive
Fluid Intake, Chronic Kidney Disease, Certain Medications, Psychogenic Polydipsia, Primary
Polydipsia, and Malabsorption Syndromes
ii) Deep amber colour or dark yellow due to good muscular exercise, high grade fever, Dehydration,
Hematuria (Blood in Urine), Liver Disorders, Hemolysis (Breakdown of Red Blood Cells), Bile Duct
Obstruction, Certain Medications, Rhabdomyolysis,
iii) Orange colour due to increased urobilinogen, concentrated urine, Vitamins and Supplements,
Hematuria (Blood in Urine), Liver or Bile Duct Disorders, Urinary Tract Infection (UTI), Porphyria,
Gilbert's Syndrome

10. iv) Smoky urine due to small amount of blood, administration of vitamin B12, aniline dye, Trauma or
Injury, Interstitial Cystitis, Systemic Diseases, Medications, Glomerulonephritis, Kidney Stones, Urinary
Tract Infection (UTI).
v) Red due to haematuria, haemoglobinuria, Inflammatory Conditions, Cancer, Medications, Blood
Disorders, Enlarged Prostate or Prostate Infection, Injury or Trauma, Bladder or Kidney Infections, Kidney
Stones, Urinary Tract Infection (UTI).
vi) Brown due to bile, Hemolysis (Breakdown of Red Blood Cells), Ingestion of Certain Foods, and
Hematuria (Blood in Urine),
vii) Milky due to pus, fat.
viii) Green due to putrefied sample, phenol poisoning, (Certain medications or dyes (e.g., methylene blue,
amitriptyline), Ingestion of certain foods or drinks.

11. APPEARANCE:
Urine is normally clear, and the presence of particulate material in a
specimen warrants further investigation
Chyluria.
-a rare condition in which the urine contains lymph.
Lipiduria.
--Fat globules appear in the urine mostly with
the nephrotic Syndrome
NORMAL
HAZY

13. ODOUR
◦ FRUITY- Ketoacidosis, starvation
◦ Mousy- Phenylketonuria
◦ Fishy – UTI with proteus, tryosinaemia
◦ Ammoniacal- uti with E.coli, long standing urine
SPECIFIC GRAVITY
Normal- 1.003-1.030
HYPOASTHENIC HYPERASTHENIC ISOASTHENIC
SG <1.007 SG >1.030 SG fixed at 1.010
Method-
1)Reagent strip
2)Refractometer
3)Urinometer
4)Falling drop method

14. Urinometer
Procedure
 Fill urinometer container 3/4th with urine.
 Insert urinometer into it so that it floats in urine without
touching the wall and bottom of container (Fig. 4.1).
 Read the graduation on the arm of urinometer at lower
urinary meniscus.
 Add or substract 0.001 from the final reading for each 3°C
above or below the calibration temperature respectively
marked on the urinometer.

15. 2. Refractometer
It measures the refractive index of urine. This procedure requires only a few drops of
urine in contrast to urinometer where approximately 100 ml of urine is required.
3. Reagent Strip Method
This method employs the use of chemical reagent strip
Significance of Specific Gravity
The normal specific gravity of urine is 1.003 to 1.030.
Low specific gravity urine occurs in:
i. Excess water intake
ii. Diabetes inspidus
High specific gravity urine is seen in:
i. Dehydration
ii. Albuminuria
iii. Glycosuria
Fixed specific gravity (1.010) urine is seen in:
i. ADH deficiency
ii. Chronic nephritis

16. CHEMICAL
EXAMINATION

17. CHEMICAL EXAMINATION
Chemical constituents frequently tested in urine are: proteins, glucose, ketones,
bile derivatives and blood.
Tests for Proteinuria
If urine is not clear, it should be filtered or centrifuged before testing. Urine may
be tested for proteinuria by qualitative tests and quantitative methods.
Qualitative Tests for Proteinuria
1. Heat and acetic acid test
2. Sulfosalicylic acid test
3. Heller’s test
4. Reagent strip method.

18. Heat and Acetic Acid Test
Heat causes coagulation of proteins. The procedure is as under:
 Take a 5 ml test tube.
 Fill 2/3rd with urine.
 Acidify by adding 10% glacial acetic acid if urine is alkaline.
 Boil upper portion for 2 minutes (lower part acts as control).
 If precipitation or turbidity appears add a few drops of 10% acetic acid.
Interpretation : If turbidity or precipitation disappears on addition of acetic
acid, it is due to phosphates; if it persists after addition of acetic acid then it is due
to proteins. Depending upon amount of protein the results are interpreted as under

19.  No cloudiness = negative.
 Cloudiness against dark background = traces (less
than 0.1 g/dl).
 Cloudiness without granularity = +(0.1 g/dl).
 Granular cloudiness=++(0.1-0.2 g/dl)
 Precipitation and flocculation = +++(0.2-0.4 g/dl).
 Thick solid precipitation = ++++ (0.5 g/dl).

22. Sulfosalicylic Acid Test
This is a very reliable test. The procedure is as under:
 Make urine acidic by adding acetic acid.
 To 2 ml of urine add a few drops (4-5) of 20% sulfosalicylic acid.
Interpretation; Appearance of turbidity which persists after heating indicates
presence of proteins.

24. Heller’s Test
 Take 2 ml of concentrated nitric acid in a test tube.
 Add urine drop by drop by the side of test tube.
Interpretation : Appearance of white ring at the junction indicates presence of
protein.

26. Reagent Strip Method
Bromophenol coated strip is dipped in urine. Change in colour of strip indicates
presence of proteins in urine and is compared with the colour chart provided for
semiquantitative grading

27. 1.Chronic Kidney Disease (CKD): Proteinuria is a common sign of kidney damage or dysfunction.
2.Diabetes Mellitus: Both type 1 and type 2 diabetes can lead to kidney damage, resulting in proteinuria.
3.Hypertension (High Blood Pressure): Uncontrolled high blood pressure can damage the kidneys and cause
proteinuria.
4.Glomerulonephritis: Inflammation of the glomeruli, the filtering units of the kidneys, can lead to proteinuria.
5.Preeclampsia: A condition that can occur during pregnancy, characterized by high blood pressure and
proteinuria.
6.Amyloidosis: A group of disorders in which abnormal proteins accumulate in tissues and organs, including
the kidneys.
7.Systemic Lupus Erythematosus (SLE): An autoimmune disease that can affect various organs, including the
kidneys, leading to proteinuria.
8.Multiple Myeloma: A type of cancer that affects plasma cells and can result in the overproduction of
abnormal proteins, some of which may end up in the urine.
9.Congestive Heart Failure: Severe heart failure can lead to decreased blood flow to the kidneys and
subsequent proteinuria.
10.Urinary Tract Infections (UTIs): In some cases, infections of the urinary tract can cause temporary
proteinuria.
Clinical Significance

28. Test for Glucosuria
1. Benedict’s Test
In this test cupric ion is reduced by glucose to cuprous oxide and a coloured precipitate is formed.
Procedure
Take 5 ml of Benedict’s qualitative reagent in a testtube.
Add 8 drops (or 0.5 ml) of urine.
Heat to boiling for 2 minutes (Fig. 4.5).
Cool in water bath or in running tap water.
Interpretation
 No change of blue colour = Negative
 Greenish colour = traces (< 0.5 g/dl)
 Green/cloudy green ppt = + (1g/dl)
 Yellow ppt = ++ (1-1.5g/dl)
 Orange ppt = +++ (1.5-2g/dl)
 Brick red ppt= ++++ (> 2g/dl)

30. 2. Reagent Strip Test
These strips are coated with glucose oxidase and the test is based on enzymatic
reaction. This test is specific for glucose. The strip is dipped in urine. If there is
change in colour of strip it indicates presence of glucose. The colour change is
matched with standard colour chart provided on the label of the reagent strip
bottle.

31. 1.Diabetes Mellitus:
1. Type 1 Diabetes: Results from the body's inability to produce insulin.
2. Type 2 Diabetes: Characterized by insulin resistance and inadequate insulin production.
2.Renal Tubular Disorders:
1. Fanconi Syndrome: A rare disorder affecting the proximal renal tubules, leading to the loss of various substances in the
urine, including glucose.
2. Renal glycosuria: A condition where the renal tubules do not reabsorb glucose properly.
3.Pregnancy:
1. Gestational diabetes can sometimes lead to transient glucosuria during pregnancy.
4.Hormonal Disorders:
1. Cushing's Syndrome: Excess cortisol in the body can lead to elevated blood glucose levels and glucosuria.
2. Hyperthyroidism: Thyroid hormone imbalances may impact glucose metabolism.
5.Medications:
1. Some medications, such as certain diuretics, can cause glucosuria.
6.Stress and Illness:
1. Severe stress or illness can cause temporary spikes in blood glucose levels and glucosuria.
7.Genetic Disorders:
1. Rare genetic disorders affecting glucose metabolism or renal function may result in glucosuria.
8.Infections:
1. Certain urinary tract infections can be associated with glucosuria.
9.Postprandial State:
1. After meals, it's normal for blood glucose levels to rise temporarily, and some glucose may spill into the urine.
Clinical Significance

32. Tests for Ketonuria
Tests for Ketonuria
1. Rothera’s test
2. Gerhardt’s test
3. Reagent strip test

33. 1. Rothera’s Test
Principle Ketone bodies(acetone and acetoacetic acid) combine with alkaline
solution of sodium nitroprusside forming purple complex.
Procedure
 Take 5 ml of urine in a test tube.
 Saturate it with solid ammonium sulphate salt.
 Add a few crystals of sodium nitroprusside and shake.
 Add liquor ammonia from the side of test tube.
Interpretation: Appearance of purple coloured ring at the junction indicates
presence of ketone bodies

35. 2. Gerhardt’s Test
It is not a very sensitive test.
Procedure
 Take 5 ml of urine in a test tube.
 Add 10% ferric chloride solution drop by drop.
 Filter it and add more ferric chloride.
Interpretation Brownish red colour indicates presence of ketone bodies

37. 3. Reagent Strip Test
These strips are coated with alkaline sodium nitroprusside. When strip is dipped in
urine it turns purple if ketone bodies are present

38. 1.Diabetic Ketoacidosis (DKA):
1. A serious complication of diabetes, particularly in cases of uncontrolled type 1 diabetes or insufficient insulin in
type 2 diabetes.
2.Starvation or Fasting:
1. Extended periods of fasting or severe calorie restriction can lead to ketonuria.
3.Low Carbohydrate Diets:
1. Diets that are very low in carbohydrates, leading the body to rely on fat metabolism for energy.
4.Alcoholism:
1. Chronic alcohol consumption can disrupt normal metabolic processes, leading to ketonuria.
5.Vomiting and Diarrhea:
1. Conditions that cause persistent vomiting or diarrhea can lead to the depletion of glucose and subsequent
ketone production.
6.Hyperthyroidism:
1. Excess thyroid hormone may increase the breakdown of fat, leading to ketonuria.
7.Pregnancy:
1. Ketonuria can occur during pregnancy, especially if there is severe nausea and vomiting (hyperemesis
gravidarum).
8.Infections and Fevers:
1. Severe infections or high fevers can increase the body's metabolic rate and lead to ketonuria.
9.Certain Medications:
1. Medications such as sodium-glucose cotransporter-2 (SGLT2) inhibitors, often used in the treatment of
diabetes, can increase the risk of ketonuria.
10.Inborn Errors of Metabolism:
1. Rare genetic disorders affecting the metabolism of fatty acids or amino acids can lead to ketonuria.
Clinical Significance

39. Test for Bile Derivatives in Urine
Tests for Bile Salts
Bile salts excreted in urine are cholic acid and chenodeoxycholic acid.
Tests for bile salts are Hay’s test and strip method.

40. 1. Hay’s Test
Principle Bile salts if present in urine lower the surface tension of the urine.
Procedure
 Fill a 50 or 100 ml beaker 2/3rd to 3/4th with urine.
 Sprinkle finely powdered sulphur powder over it.
Interpretation: If bile salts are present in the urine then sulphur powder sinks,
otherwise it floats.

42. 2. Strip Method
Coated strips can be used for detecting bile salts as for other constituents in
urine.
Causes for bile salts in urine:
i. Obstructive jaundice

43. Tests for Urobilinogen
Normally a small amount of urobilinogen is excreted in urine (4 mg/24 hr). The sample
should always be collected in a dark coloured bottle as urobilinogen gets oxidised on
exposure to light.
Tests for urobilinogen in urine are Ehrlich’s test and reagent strip test.

44. 1. Ehrlich’s Test
Principle Urobilinogen in urine combines with Ehrlich’s aldehyde reagent to give a red
purple coloured compound.
Procedure
 Take 10 ml of urine in a test tube.
 Add 1 ml of Ehrlich’s aldehyde reagent.
 Wait for 3-5 minutes.
Interpretation: Development of red purple colour indicates presence of
urobilinogen. A positive test is subsequently done in dilutions; normally it is positive in
upto 1:20 dilution.

46. 2. Reagent Strip Test
These strips are coated with p-dimethyl-aminobenzaldehyde. When strip is
dipped in urine, it turns reddish-brown if urobilinogen is present

47. 1.Diabetic Ketoacidosis (DKA):
1. A serious complication of diabetes, especially type 1 diabetes, where the body produces high
levels of ketones due to insufficient insulin.
2.Starvation or Fasting:
1. Extended periods of fasting or severe calorie restriction can lead to the breakdown of fat for
energy, resulting in ketone production.
3.Low Carbohydrate Diet:
1. Diets very low in carbohydrates can cause the body to rely on fat metabolism, leading to
ketone production.
4.Alcoholism:
1. Chronic alcohol consumption can disrupt normal metabolism, leading to increased ketone
production.
5.Prolonged Exercise:
1. Intense and prolonged physical activity, especially in the absence of sufficient carbohydrate
intake, can result in ketonuria.
6.Hyperthyroidism:
1. Overactive thyroid function can increase the breakdown of fat, leading to ketone production.
7.Febrile Illness:
1. High fever and certain infections can cause increased metabolism and ketonuria.
Clinical Significance

48. 1.
Malnutrition:
1. Inadequate intake of essential nutrients, including carbohydrates, can lead to ketone
production.
2.Glycogen Storage Disorders:
1. Genetic disorders that affect the storage and release of glycogen can lead to ketosis.
3.Inborn Errors of Metabolism:
1. Certain genetic disorders affecting the metabolism of fats and carbohydrates can lead to
ketonuria.
4.Pregnancy:
1. During pregnancy, especially in the first trimester, some women may experience mild
ketonuria.
5.Severe Stress or Trauma:
1. Critical illness, major surgery, or trauma can lead to increased stress on the body, resulting in
ketonuria.
6.Certain Medications:
1. Medications that affect carbohydrate metabolism or insulin function may contribute to
ketonuria.
7.Anorexia Nervosa:
1. Individuals with this eating disorder may experience ketonuria due to severe caloric restriction.
8.Reye's Syndrome:
1. A rare but serious condition that can occur in children recovering from viral infections,

49. Tests for Bilirubin (Bile Pigment) in Urine
Bilirubin is breakdown product of haemoglobin. Normally no bilirubin is passed in
urine. Following tests are done for detection of bilirubin in urine:
1. Fouchet’s test
2. Foam test
3. Reagent strip test

50. 1. Fouchet’s Test
Principle Ferric chloride oxidises bilirubin to green biliverdin.
Procedure
 Take 10 ml of urine in a test tube.
 Add 3-5 ml of 10% barium chloride.
 Filter through filter paper.
 To the precipitate on filter paper, add a few drops of
 Fouchet’s reagent (ferric chloride + trichloroacetic acid).
Interpretation: Development of green colour indicates bilirubin.

52. 2. Foam Test
It is a non-specific test.
Procedure
 Take 5/10 ml of urine in a test tube.
 Shake it vigorously.
Interpretation: Presence of yellow foam at the top indicates presence of
biluribin.
3. Reagent Strip Test
Principle It is based on coupling reaction of bilirubin with diazonium salt
with which strip is coated. Dip the strip in urine; if it changes to blue colour
then bilirubin is present

53. Liver Diseases:
•Hepatitis (inflammation of the liver)
•Cirrhosis (scarring of the liver)
•Liver infections
Hemolytic Anemias:
• Sickle cell anemia
• Thalassemia
• Autoimmune hemolytic anemia
Biliary Tract Obstruction:
•Gallstones blocking the bile ducts
•Tumors in the liver or bile ducts
1.Gilbert's Syndrome:
1. A genetic condition that affects the way the liver processes bilirubin, leading to mild
increases in bilirubin levels.
2.Cirrhosis:
1. Advanced scarring of the liver tissue, often due to chronic liver diseases.
3.Hemolysis:
1. Any condition that causes the destruction of red blood cells, releasing bilirubin into the
bloodstream.
4.Certain Medications:
1. Drugs that can affect the liver, such as certain antibiotics, antivirals, and anti-seizure
medications.
Clinical Significance

54. 1.Infectious Diseases:
1. Some infections affecting the liver, such as infectious hepatitis.
2.Liver Tumors:
1. Tumors in the liver can disrupt normal liver function and increase bilirubin levels.
3.Hepatotoxicity:
1. Exposure to substances toxic to the liver, such as certain chemicals and drugs.
4.Hemorrhage:
1. Severe bleeding or hemorrhage can lead to an increase in bilirubin levels.
5.Congestive Heart Failure:
1. In some cases, heart failure can lead to liver congestion and impaired liver function.
6.Sepsis:
1. Severe infection that can affect multiple organs, including the liver.
7.Inflammatory Bowel Disease:
1. Conditions like Crohn's disease and ulcerative colitis can affect the liver and bile ducts.
8.Genetic Disorders:
1. Certain genetic disorders affecting liver function can lead to increased urobilinogen.

55. Tests for Blood in Urine
Tests for detection of blood in urine are as under :
1. Benzidine test
2. Orthotoluidine test
3. Reagent strip test

56. 1. Benzidine Test
Procedure
 Take 2 ml of urine in a test tube.
 Add 2 ml of saturated solution of benzidine with glacial acetic acid.
 Add 1 ml of H2O2 to it.
Interpretation Appearance of blue colour indicates presence of blood.
Benzidine is, however, carcinogenic and this test is not commonly used.

57. 2. Orthotoluidine Test
Procedure
 Take 2 ml of urine in a test tube.
 Add a solution of 1 ml of orthotoluidine in glacial acetic acid.
 Add a few drops of H2O2.
Interpretation Blue or green colour indicates presence of blood in urine.
3. Reagent Strip Test
The reagent strip is coated with orthotoluidine. Dip the strip in urine. If it changes
to blue colour then blood is present

58. 1.Urinary Tract Infections (UTIs): Infections in the urinary tract, such as in the bladder or kidneys, can
lead to hematuria.
2.Kidney Stones: Hard deposits that form in the kidneys can cause bleeding as they move through the
urinary tract.
3.Bladder or Kidney Infections: Infections in the bladder or kidneys may cause inflammation and
bleeding.
4.Injuries or Trauma: Trauma to the urinary tract, such as a blow to the kidneys or bladder, can result in
blood in the urine.
5.Enlarged Prostate: In men, an enlarged prostate can lead to hematuria.
6.Kidney Disease: Various kidney diseases, including glomerulonephritis and polycystic kidney disease,
can cause blood in the urine.
7.Bladder or Kidney Cancer: Tumors in the bladder or kidneys may lead to hematuria.
Clinical Significance

59. 1.Strenuous Exercise: Intense physical activity, such as long-distance running, can cause microscopic
blood in the urine, known as exercise-induced hematuria.
2.Blood Disorders: Conditions like sickle cell anemia and hemophilia can cause bleeding throughout the
body, including the urinary tract.
3.Medications: Certain medications, such as blood thinners, can increase the risk of bleeding.
4.Inherited Conditions: Some individuals may have a genetic predisposition to kidney or bladder
problems that can lead to hematuria.
5.Vigorous Sexual Activity: In some cases, vigorous sexual activity may cause microscopic blood in the urine.
6.Irritation of the Urethra: Irritation of the urethra due to sexual activity, catheter use, or other reasons
can result in hematuria.
7.Interstitial Cystitis: A chronic condition causing inflammation of the bladder wall may lead to blood in
the urine.
8.Blood Clotting Disorders: Disorders that affect blood clotting may result in bleeding in various parts of
the body, including the urinary tract

60. Microscopic examination of urine
Identify the following formed elements found in urine sediment, and discuss their clinical significance:
1. • Bacteria
2. • Clue cells
3. • Fat
4. • Fecal contaminants
5. • Fibers
6. • Hemosiderin
7. • Mucus threads
8. • Parasites
9. • Spermatozoa
10. • Starch
11. • Trichomonads
12. • Ye a s t

61. OBSERVATIONS
ORGANISED ELEMENTS UNORGANISED ELEMENTS
CELLS : RBCs ; WBCs ; Epithelial cells ; Oval fat bodies are Crystalline & amorphous substances
CASTS : CELLULAR ACELLULAR
- RBC cast -Hyaline cast
- WBC cast - Granular cast
-Epithelial cast -Waxy cast
-Fatty cast
ORGANISMS : Bacteria ; Fungi ; Parasite
SPECIMEN
Preferred specimen is –
- freshly voided, midstream , clean catch , first morning sample.

62. COLLECTION OF SAMPLE
• Early morning sample is the best specimen.
• It provides an acidic and concentrated sample which preserves the formed elements (RBCs, WBCs and
casts) which otherwise tend to lyse in a hypotonic or alkaline urine.
• The specimen should be examined fresh or within 1-2 hours of collection.
PREPARATION OF SEDIMENT
• Take 5-10 ml of urine in a centrifuge tube.
• Centrifuge for 5 minutes at 3000 rpm.
• Discard the supernatant.
• Resuspend the deposit in a few ml of urine left.
• Place a drop of this on a clean glass slide.
• Place a coverslip over it and examine it under the microscope.

63. SPECIMEN
Preferred specimen is –
- freshly voided, midstream , clean catch , first morning sample.

64. EXAMINATION OF SEDIMENT
Visualization Techniques
Sternheimer-Malbin
• A supravital stain that characteristically stains cellular structures and other formed elements
• Enables detailed viewing and differentiation of cells, cast inclusions, and low-refractile elements (e.g.,
hyaline casts, mucus)
0.5% toluidine blue
• A metachromatic stain that enhances the nuclear detail of cells
• Aids in differentiating WBCs and renal tubular epithelial cells
2% acetic acid
• Accentuates the nuclei of leukocytes and epithelial cells
• Lyses RBCs

65. Fat stains: Sudan III, il red O
• Stains triglyceride (neutral fat) droplets a characteristic orange (Sudan III) or red (oil red O) color
• Used to confirm the presence of fat in urine
Gram stain
• Identifies and classifies bacteria as Gram negative or Gram positive
• Aids in the identification of bacterial and fungal casts
Prussian blue reaction
• Identifies hemosiderin, which can be free floating, in epithelial cells, or in casts
Hansel stain
• Aids in the identification of eosinophils
Cytokeratin Immunostaining
•Immunostaining for cytokeratins can be performed to detect specific proteins in urine cytology, aiding in
the identification of malignant cells.

66. Periodic Acid-Schiff (PAS) Stain:
•PAS stain is used to highlight the presence of glycogen and other carbohydrates in cells, and it can be
applied in urine cytology to identify certain cellular components.
Methylene Blue Stain:
•Methylene blue is a basic dye that can be used to stain cells in urine, providing contrast and aiding in the
identification of cellular features.
Feulgen Stain:
•Feulgen stain is specific for DNA and is used to visualize the amount of DNA in cell nuclei. It can be
employed in urine cytology to assess nuclear characteristics.
Diff-Quik Stain:
•Diff-Quik is a rapid differential stain often used in clinical laboratories for urine cytology. It allows for quick
visualization of cellular details.
Romanowsky Stains (e.g., Wright's Stain, Giemsa Stain):
•These stains are versatile and commonly used in urine cytology to provide good differentiation of cellular
components.

67. Microscopic Techniques
Phase contrast microscopy
• Enhances the imaging of translucent or low-refractile formed elements
Interference contrast microscopy
• Enhances the imaging of formed elements by producing three-dimensional images
Polarizing microscopy
• Used to confirm the presence of cholesterol droplets by their characteristic Maltese cross pattern
• Aids in the identification of crystals

68. White blood cells (neutrophils)
stained with 0.5%
toluidine blue. Brightfield, ×400.
Oval fat body stained with Sudan I
II stain. Note the characteristic
orange-red coloration of neutral
fat (triglyceride) droplets. Brightfield, ×400
Eosinophil (arrow) in urine stained
with Hansel stain. Cytospin, ×400.
Bacteria. Gram stain of Gram-negative
rods and Gram-positive cocci. Brightfield,
×1000.
Urine cytology (Papanicolau stain)
from July 2010 showing large numbers
of neutrophil polymorphs together with
scattered urothelial cells

69. CELLS
ERYTHROCYTES
Under high power objective, they appear normally as :
- Pale biconcave disc shaped cells without nucleus or granules
- Size- 7.2 microns diameters.
- NORMAL (0-2 CELLS/HPF)
In hypertonic urine : Crenated RBCs with spikey surface.
In hypotonic urine : swollen RBCs.
CONFUSERS : a) Oil droplets
b) Yeast cells

70. CAUSES OF RBCS IN URINE
RENAL DISEASE LOWER URINARY TRACT EXTRARENAL DISEASE TOXIC
REACTIONS
GLOMERULONEPHRITIS INFECTION APPENDICITIS DRUGS
LUPUS NEPHRITIS CALCULUS SALPINGITIS (SULFONAMIDE,
INTERSTITIAL NEPHRITIS TUMOR DIVERTICULITIS SALICYLATE ETC)
STRICTURE MALARIA ANTICOAGULANT
HAEMORRHAGIC CYSTITIS SBE THERAPY
PAN
MALIGNANT HYPERTENSION
TUMORS OF RECTUM, COLON
DYSMORPHIC ERYTHROCYTES
-RBC’s with cellular protusions or fragmantation
- >80% Dysmorphic cellS – glomerular pathology SUSPECTED.

71. Forms of Red Blood Cells in Urine
Normal cell
Burr cell
Ghost cell
Cell with thin membrane and
without hemoglobin
canthocyte Target cell
Stomatocyte
Schistocyte
Dysmorphic (schizocyte,
crenated/echinocyte) and
normal forms of red blood cells.

72. LEUCOCYTES
- Neutrophil is the predominant type of leukocyte
that appears in urine.
- Under high power objective,leucocytes are
seen as :
round cells with granules inside with mutilobated
nuclei (10-15 ) microns in size.
- Normal : ( 1-2 ) pus cells /HPF.
- Presence of pus cells more than 30cells/HPF or in
clumps signifies acute infection
CONFUSER : Small epithelial cells.

73. White Blood Cells: Microscopic Features and Correlations
Microscopic features
Neutrophils
• Spherical cells, 12–16 μm in diameter
• Granular cytoplasm
• Lobed nuclei
• Glitter cells—dilute urine (low SG)
Lymphocytes
• Spherical cells, 6–9 μm in diameter
• Mononuclear
Monocytes and macrophages
• Spherical cells, 20–25 μm in diameter
• Granular cytoplasm
• Mononuclear
• Cytoplasm often vacuolated with ingested debris

74. Eosinophils T
- Not normally seen in urine, and the finding > 1%
eosinophils among leukocyte is considered signifcant .
- Seen in-tubulointerstitial disease
allergic interstitial nephritis
Lymphocytes and Mononuclear Leukocytes.
- Small lymphocytes and histiocytes are normally present in urine
- When mononuclear cells constitute >30% or more of a differential
count, chronic inflammation is indicated.

75. Look-alike elements
• Renal tubular epithelial cells (collecting duct cells)
• Dead trichomonads
• Crenated RBCs
Correlation with physical and chemical examinations
• Leukocyte esterase reaction—can be negative despite increased WBCs owing to excess
hydration or when the WBCs are lymphocytes
• Negative nitrite reaction: suggestive of inflammation or nonbacterial infection
• Positive nitrite reaction: suggests bacterial infection
Two renal collecting duct cells. Their polygonal shape and nuclear detail
distinguish them from leukocytes. Brightfield, ×400

76. A clump of white blood cells. One red
blood cell and budding yeast are also
present. Brightfield, Sedi-Stain, ×400
Degenerating white blood cells with the
formation of blebs. Phase contrast, ×400
Several white blood cells with characteristic
cytoplasmic granules and lobed nuclei
surrounding a squamous epithelial cell.
Budding yeast cells are also present.
Brightfield, Sedi-Stain,×400.
Cytospin of urine sediment. Several white
blood cells with cytoplasmic vacuoles and
blebs in urine. Note that intracellular (arrow)
and extracellular bacteria are present. Wright
stained, ×1000.

77. Formation of myelin filaments in
disintegrating white blood cells. Phase
contrast, ×400
Lymphocyte (arrow) in a cytospin of
urine sediment; Wright stain. Brightfield,
×400.
Macrophages and several other white blood cells. (A) Brightfield,
×400. (B) Brightfield, Sedi-Stain, ×400.

78. EPITHELIAL CELLS
SQUAMOUS EPITHELIAL CELLS TRANSITIONAL CELLS RENAL TUBULAR CELLS LIPIDS
&
PIGMENTS IN
TUBULAR EPITHELIAL
CELLS
Squamous epithelial cells :
- They line the distal 1/3RD urethra & vagina.
- They are seen as large , rectangular ,
flattened cells with abundant cytoplasm,
small , central, round nucleus.

79. Transitional epithelial cells :
-They line the urinary tract from the renal pelvis to the lower third of the urethra.
- They are round or pear shaped, with a round, centrally located nucleus
-When stained, transitional cells have dark blue nuclei with pale blue
cytoplasm .
- transitional cells have a characteristic endo-ecto cytoplasmic rim. .

80. Renal Tubular Epithelial Cells
-These are the most signifcant types of epithelial cells found in urine because the finding
of an increased number indicates tubular damage.
Renal epithelial cells from PCT &DCT: Renal epithelial cells from CD:
singly and are large (14–60 µm), oblong cuboidal or polygonal shape
with coarse granular eosinophilic cytoplasm. basophilic endo-ecto cytoplasmic rim
Nuclei are small with dense chromatin. Large and eccentric nucleus
seen in- acute tubular necrosis seen in-renal transplant rejection, drug
or heavy metal toxicities. acute tubular necrosis

81. Oval fat bodies :
These are degenerated renal tubular epithelial cells filled with highly refractile
lipid (cholesterol) droplets.
They are seen in nephrotic syndrome .
Pigment in Renal Tubular Epithelial Cells
1)Haemoglobin/myoglobin pigments
2)Melanin pigment
3)Bilirubin pigment