AUBF_Midterms_Lec: Microscopic examination

Macroscopic screening: Color

Microscopic Correlations:

Microscopic Correlations: Blood

Macroscopic screening: Clarity

Microscopic Correlations:

Microscopic Correlations: Hematuria vs. hemoglobinuria/myoglobinuria

Macroscopic screening: Blood

Microscopic Correlations:

Microscopic Correlations: RBCs, and RBC Casts

Macroscopic screening: Protein

Microscopic Correlations:

Microscopic Correlations: Casts, Cells

Macroscopic screening: Nitrite

Microscopic Correlations:

Microscopic Correlations: Bacteria, WBCs

Macroscopic screening: Leukocyte esterase

Microscopic Correlations:

Microscopic Correlations: WBCs, WBS Cast, bacteria

Macroscopic screening: Glucose

Microscopic Correlations:

Microscopic Correlations: Yeast

Thomas Addis

• he developed a quantitative method of examining urine sediment

Hemocytometer

•12-hr specimen

•RBCs, WBCs, casts and epithelial cells

• Used to primarily monitor the course of diagnosed cases of renal disease

Preparation of the Urine Sediment, give criterias

• Freshly voided urine

• Midstream clean-catch specimen

• 10-15 mL (12 mL)

Centrifugation of urine

• Centrifuge for 5mins @ 400 RCF

mL of urine sediment that was left after decantation

• 0.5-1.0mL

Explain examination of urine sediment

• observe at 10 fields (both LPF and HPF)

Sternheimer-Malbin

Stain: WBC, Epithelial Cells, Casts

Sternheimer-Malbin Composition

• Crystal Violet

• Safranin O

Toluidine Blue

Stain:

• enhances nuclear detail

• differentiate WBC from RTE cells

2% Acetic Acid

Stain:

• Lyses RBC and enhances nuclei of WBCs

• Distinguish RBC from WBCs, yeast, oil droplets, and crystals

Lipid stains

• Oil red O

• Sudan III

Stain:

• TAGs

• Identify fat droplets and lipid-containing cells, and casts

Gram stain

Stain:

• Diff. gram pos and gram neg bacteri

• Identify bacterial casts

Hansel Stain

Stain:

• uses Meth Blue and Eosin Y to stain eosinophilic granules

Acute Interstitial Nephritis (AIN)

• This disease is determined thru the use of Hansel Stain

Prussian Blue stain

Stain:

• identify yellow-brown granules of hemosiderin in cells and casts

Brightfield microscopy

• common microsopic technique use droutine urinalssi

Phase-contrast microscopy

• this microscopic technique is best used for low refractive indices

  • Hyaline cast

  • mixed cellular casts

  • mucus threads

  • Trichomonas vaginalis

Polarized light microscopy

• Cholesterol

• microscopic technique to check form Maltese cross formation which indicates what metabolite

Type of microscopy used depends on (give 3)

• Spx type

• Refractive index of object

• Imagery of unstained living cells

Fluorescence microsocpy

Interference-contrast

RBC Appearance in hypertonic urine

Crenated apperance

RBC Appearance in hypotonic urine

Ghost cells

Appearance of RBC with glomerular membrane damage

Dysmorphic appearance

Sources of identification error for RBCs

• Yeast cells

• Oil droplets

• Air bubbles

RBCs

• These sediments appear with conditions such as glomerulonephritis and strenuous exercise

Correlations with RBCs

• Color

• Reagents strip blood reaction

WBCs: in hypotonic urne

Glitter cells

WBC: Sources of identification error

Renal tubular epithelial cells

WBC: Urinalysis correlation

• LE

• Nitrite

• Specific Gravity

• pH

Squamous Epithelial cells

• Largest cells that can be seen in microscopic exam

Squamous Epithelial cells

• These are the cells used for point of reference whenever we do microscopic examination

increased no. of SEC inidcates what?

• contam of urine specimen due to poor collection technique

RTE Cells

• Leukocyte esterase and nitrite (pyelonephritis)

• Color

• Clarity

• Protein Bilirubin (hepatitis)

• Blood

Transitional cells: Urinalysis correlations

• Clarity

• blood (malignancy)

Transitional cells: Clinical significane

increased nos. indicate

• UTI

• Renal carcinoma

• catheterization

RTE Cells: Appearance

• Rectangular, columnar, round, oval, or cuboidal with an eccentric nucleus possibly bilirubin stained or hemosiderin laden

RTE Cells: Clinical significane

• tubular necrosis

(to add more)

Oval Fat Bodies: Urinalysis correlations

• Clarity

• Blood

• Protein

• Free fat droplets/Fatty Casts

Oval Fat bodies: Clinical significance

• Increased in glomerular damage by:

  • nephrotic syndrome,

  • tubular necrosis,

  • diabetes mellitus, long bone trauma

Bacteria: Appearance

• Small spherical and rod-shaped structures

Bacteria: ID errors

• Amorphous phosphates

• Amorphous urates

How are Bacteria reported in urinalysis

• R,F,Mod,Many (HPF)

note: Presence of WBCs may be required

Bacteria: Urinalysis correlation

• pH

• Nitrite

• LE

• WBCs

Yeast: Apperance

Small, oval, refractile structures with buds and/or mycelia

Yeast: ID Error

• RBCs

Yeast: Correlation

Correlated with:

• Glucose

• LE

• WBCs

Parasites (trichomonas)

• Pear-shaped, motile, flagellated

Parasites: ID error

Errors:

• WBCs

• RTE cells

Parasites (Trichomonas): Correlation

Correlation

• LE

• WBCs

Enterobius vermicularis ova

This parasite is accidentally found due to fecal contamination

Spermatozoa: appearance

Tapered oval head with long, thin tail

not required

spermatozoa is (required/not required) to be reported based on lab protocols

Spermatozoa: correlation

Corrleation

• Protein

Mucus threads

• Single or clumped threads with a low refractive index

Mucus threads: ID Error

Error

• Hyaline Casts

How can we differentiate RTE cells and WBC?

Based on the presence of protein in Chem exam

• RTE Cells = (+) Proteins

• WBC = (-) Proteins

Where are urine casts formed

Lumen of Distal Convoluted tubule and collecting duct

Casts are usually observe in what objective? Manner of reporting?

• LPF detected around the cover slip edge

Tamm-Horsfall protein/Uromodulin

• these are proteins that are responsible in forming casts

• released by the cells in the Renal Tubules

•Substances that can become embedded in casts include (give 5)

• Cells

• Bacteria

• Granules

• Pigments

• Crystals

Hyaline Cast

_:Appearance

Colorless homogenous matrix

Hyaline Cast

_:ID Error

Mucus, fibers, hair, increased lighting

Hyaline Cast

_: Correlation

• Protein  -Blood (exercise)  • Color (exercise)

Hyaline Cast

_:Clinical significance

Glomerulonephritis  Pyelonephritis  Chronic renal disease Congestive heart failure (CHF) Stress and exercise

RBC Cast

_:Appearance

Orange-red color, cast matrix containing RBCs

RBC Cast

_:ID Error

RBC clumps

RBC Cast

_: Correlation

A cast: • RBCs  • Blood  • Protein

RBC Cast

_:Clinical significance

A cast: Glomerulonephritis  Strenuous exercise

WBC Cast

_:Appearance

Cast matrix containing WBCs

WBC Cast

_:ID Error

A cast: WBC clumps

WBC Cast

_: Correlation

A cast: WBCs Protein LE

WBC Cast

_:Clinical significance

A cast: Pyelonephritis Acute interstitial nephritis (eosinophils)

Bacterial Casts

_:Appearance

A cast:  Bacilli bound to protein matrix

Bacterial Casts

_:ID Error

A cast: Granular casts

Bacterial Casts

_: Correlation

A cast: WBC Cast LE Protein WBCs Nitrite Bacteria

Bacterial Casts

_:Clinical significance

A cast: Pyelonephritis

Epithelial Cell Cast

_:Appearance

RTE cells attached to protein matrix

Epithelial Cell Cast

_:ID Error

A cast:  WBC cast

Epithelial Cell Cast

_: Correlation

A cast:  Protein RTE cells

Epithelial Cell Cast

_:Clinical significance

A cast: Renal tubular damage

Fatty Cast

_:Appearance

Fat droplets and oval fat bodies attached to protein matrix

Fatty Cast

_:ID Error

A cast: Fecal debris

Fatty Cast

_: Correlation

A cast: • Protein  • Free fat droplets  • Oval fat bodies

Fatty Cast

_:Clinical significance

A cast: Nephrotic syndrome Toxic tubular necrosis Diabetes mellitus Crush injuries

Granular Cast

_:Appearance

Coarse and fine granules in a cast matrix

Granular Cast

_:ID Error

A cast: Clumps of small crystals Columnar RTE cells

Granular Cast

_: Correlation

A cast: Protein Cellular Casts RBCs and WBCs

Granular Cast

_:Clinical significance

A cast: Glomerulonephritis Pyelonephritis Stress and exercise

Waxy Cast

_:Appearance

Highly refractile cast with jagged ends and notches

Waxy Cast

_:ID Error

A cast:  Fibers and fecal material

Waxy Cast

_: Correlation

A cast: Protein Cellular Casts Granular casts RBCs and WBCs

Waxy Cast

_:Clinical significance

A cast: Stasis of urine flow  Chronic renal failure