AUBF: MICROSCOPIC EXAMINATION

10- 15mL.

Standard amount of urine

12 ml

volume is frequently used

5 minutes at 400(RCF)

centriguation of urine

0.5mL-1mL

volume consider in sendiment preparation

Concentration factor

relates to the probability of detecting elements present in low quantities and is used when quantitating the number of elements present per milliliter

rather than poured of

urine should be aspirated of

Vigorous agitation

should be avoided, as it may disrupt some cellular elements.

20µL (0.02ml)

Conventional glass-slide method

: 22x22mm

glass cover slip

10 fields.

Examine both LPF and HPF minimum of

LPF

objectives use in Cast and epithelial cells

HPF

obejctives use in WBC, RBC, yeast cell

sternheimer malbin

delinates structire and contrasting colors of nucleus and cytoplasm

toluidine blue

enhances nucleur details

2% acetic acid

lyses the RBC and enhnaces of ebc

lipids: sudan 3 and oil red o

strains triglycerided and neutral fats

gram stain

uses in differentiating gram neg to pos

hansel stains

methrlene blue and eosin y stains the eosinphilic granules

prussian blue

stains structure containing iron

Cytodiagnostic Urine Testing

It is done using a cytocentrifuge followed by staining with Papanicolaou stain

Cytodiagnostic Urine Testing

detecting and monitoring renal disease

→ Detection of malignancies of the lower urinary tract

Cytodiagnostic Urine Testing

Provide information about renal tubular changes associated w/ transplant rejection. → Diagnose viral, fungal, and parasitic infections

Cytodiagnostic Urine Testing

Analysis of cellular inclusions, pathologic casts and inflammatory conditions

Specimen: FIRST MORNING SPECIMEN

bright field

routinely uses

phase microscope

visualization of elemenets with low refractive indices: trichomonas

polarizing microscope

identification of cholesterol on oval fats

dark field microscope

identifaction of treponema

flourescence microscopy

naturally flourescent microgoorganism: labeled antigens and antibody

inteference microscope

three dimensional microscopy image

Red Blood Cells

Smooth, non-nucleated, biconcave disks

Concentrated: shrink diluted urine:

Red Blood Cells

adding of acetic acid to apportion of the sediment can be helpful in identification

DYSMORPHIC

RBC's that vary in size, cellular protrusion, or are fragmented are termed as

Red Blood Cells

associated with:

glomerular membrane.

Vascular injury w/in the genitourinary tract.

Strenuous exercise

Menstrual contamination

White Blood Cells

Larger than RBC

Neutrophils

→ Predominant WBC in the urine.

Glitter cells

arge neutrophils that produces a sparkling appearance

Eosinophil

drug induce interstitial nephritis.

Eosinophil

Small amount seen in UTI and renal transplant rejection

Eosinophil

Hansel stain and Wright stain can be used for differentiation.

Lymphocytes

Smallest WBC, resembles as RBC

Lymphocytes

Seen in early stage of renal transplant rejection

Monocytes, Macrophages and Histiocytes

Appear vacuolated or contain inclusions

Oval Fat Bodies

Examine using fat stains under polarized microscopy.

Oval Fat Bodies

Lipid containing RTE cells

Oval Fat Bodies

most clinically significant epithelial cells.

Oval Fat Bodies

Increase amount is indicative of necrosis of the renal tubule with the possibility of affecting the overall renal function

Bubble cells

RTE cells containing large nonlipid filled vacuoles

Bacteria

Not normally present in the urine. → Present in the form of cocci or bacilli.

Presence of bacteria can be indicative of either upper or lower UT

Enterobacteriaceae (Gram(-)rods).

Most frequently bacteria

Staphylococcus and Enterococcus

are also capable of causing UTI

Yeast

Appear as small, refractive oval structures that may or may not contain a bud.

Candida albicans are seen in diabetic, immunocompromised patients and women with vaginal moniliasis.

true yeast infection

should be accompanied by the presence of WBC

Parasites T

Most frequently encountered parasite in the urine

Trichomonas vaginalis

Sexually transmitted pathogen associated w/ vaginal inflammation.

Trichomonas vaginalis

a pear-shaped flagellate with an undulating membrane.

Trichomonas vaginalis

Darting movement

Schistosoma haematobium

Bladder parasite → Seldom seen in the US.

Enterobius vermicularis

Most common urine contaminant.

Spermatozoa

Appear as Oval, slightly tapered heads and long, flagella like tails.

Spermatozoa

Clinically significant in cases of male infertility or retrograde ejaculation

Spermatozoa

Positive in chemical test for protein when increased amount

Mucus

Protein material produced by the glands and epithelial cells of the lower genitourinary tract and RTE cells.

Mucus

Major constituent is Tamm-Horsfall Protein.

Mucus

Appears as thread-like structure

Mucus

Frequently seen in female

Casts

only urinary sediment that are unique to the kidney

Cast

Their shape is the representative of the tubular lumen.

Cast

It has a parallel side, rounded ends and may contain other elements present in filtrate.

Cast matrix

dissolve easily in dilute alkaline urine

Tamm-Horsfall protein.

Major constituent of casts is

Step by step formation of Tamm Horsfall protein matrix.

Aggregation o Formation of loose fibrillar network o Additional network to form a solid structure. o Attachment of urinary constituent o Detachment from the epithelial cell o Excretion of cast

cylinduria

The presence of urinary cast is termed

Hyaline casts

Most frequently seen casts

Hyaline casts

Colorless → Stained w/ Sternheimer-Malbin

Hyaline casts

Parallel sides and rounded ends. → Cylindroid forms

Hyaline casts

Wrinkled or convoluted shapes indicates aging of the cast matrix

RBC casts

much more specific showing bleeding w/in the nephron

RBC casts

Primarily associated w/ glomerulonephritis → Seen also in individuals participating in strenuous

RBC casts

Detected under LPO by their orange-red color

RBC casts

They are more fragile, irregular shape and may exists as fragments.

RBC casts

In the presence of massive hemoglobinuria or myoglobinuria, homogenous orange-red or red brown casts may be observed

WBC casts

Presence signifies infection or inflammation w/in the nephron.

Bacterial casts

Commonly seen in pyelonephritis.

Bacterial casts

Resembles as granular casts

Bacterial casts

Considered when increase wbc, bacteria and wbc casts are seen

Bacterial casts

Confirmatory: Gram stain