CLINICAL MICROSCOPY  Arteries and Veins – where wastes will be
 tests body fluids but mainly involved in the
study of urine (most common body fluid)
 Urine – most common specimen that is used in
determining diseases in man
URINARY SYSTEM
 consists of two bean-shaped kidneys, the
ureters, which carry urine into the bladder for
storage, and the urethra, which transports
urine outside the body
KIDNEYS
 maintain homeostasis by regulating:
1. fluid balance – maintain 60% water in our
body by preventing excessive release; or
release more if needed
2. acid-base balance – for metabolism
3. electrolyte balance – most common
component of urine are electrolytes
 primarily excrete waste products and serve to
maintain blood pressure and erythropoiesis
(production of RBCs)
 each kidney contains 1 to 1.5 million nephrons;
these are the functional units of the kidney
 important part of the kidneys because this is
where urine is being formed
 Nephron – composed of a glomerulus, which is
the filtering unit, and renal tubules, which are 30
to 40 mm in length
 Glomerulus enclosed by glomerular capsule
(or Bowman’s capsule) – where filtration
happens
 Renal Tubules – where the wastes and urine
will be collected
carried from different parts of the body going to
kidneys for filtration
URINE FORMATION
 The kidneys filter unwanted substances from
the blood and produce urine to excrete them.
These processes ensure that only waste and
excess water are removed from the body.
1. Glomerular Filtration
 each nephron has a glomerulus, the site of
blood filtration
 coming from afferent arteriole, blood
with wastes will be coming in and will be
filter in the glomerulus
 glomerulus is a network of capillaries
surrounded by a cuplike structure, the
glomerular capsule (or Bowman’s
capsule)
 As blood flows through the glomerulus,
blood pressure pushes water and solutes
from the capillaries into the capsule through
a filtration membrane
 Principle of filtration is size
 Filtrates – small particles (water) that
passed through
2. Tubular Reabsorption
 not all those that are filtered will be brought
for excretion, some of it is still important,
that’s why there is a need for reabsorption
 when the ultrafiltrate enters the proximal
convoluted tubules (PCT), cellular transport
mechanisms begin to reabsorb essential
substances and water
 ultrafiltrate – those that were filtered
from glomerulus
 Cellular mechanisms involved in tubular
reabsorption can be active or passive
transport because of the differences in
higher and lower concentrations.
 active transport – need energy; lower
to higher concentration
 passive transport – no need for
energy; higher to lower concentration
Substances reabsorbed by active transport
include:
 glucose, amino acids, and salts in the
PCT
 chloride in the ascending loop of Henle
 sodium in the distal convoluted tubules
Passive transport
 The ascending loop of Henle is
impermeable to water; therefore, passive
reabsorption of water takes place in all other
parts of the nephron.
 water could be reabsorb in any other
part of nephron except in ascending
loop of Henle
 Urea is passively reabsorbed in the PCT
and the ascending loop of Henle
 Sodium is passively reabsorbed in the
ascending loop of Henle
3. Secretion
 The filtrate absorbed in the glomerulus flows
through the renal tubule, where nutrients
and water are reabsorbed into capillaries.
  At the same time, waste ions and 2. Random Urine – collected any time of the day
hydrogen ions pass from the capillaries for routine urinalysis
into the renal tubule.  most common specimen collection for urine
 Capillaries – exchange of blood 3. Fasting or Post-prandial Urine – used for
(oxygenated red; deoxygenated red) glucose determination
 Secretion – release of substances from 4. Timed Urine – collection of all urine output
blood vessels going to tubules within 12 hours or 24 hours for clearance tests
 The secreted ions combine with the  Clearance Test – measure how long it will
remaining filtrate and become urine. take for your body to remove all chemicals
 The urine flows out of the nephron tubule that is being tested
into a collecting duct. It passes out of the
kidney through the renal pelvis, into the METHODS OF COLLECTION
ureter, and down to the bladder. 1. Clean midstream catch – urinate a small
amount into the toilet bowl, then stop the flow of
urine, then get urine sample.
COMPOSITION OF URINE  Why? because there are microorganism
 Water – largest component of urine that could be present specially in the
 Urea – accounts for half of the total dissolved urethra, that’s why the first urine that will be
solids in urine released will carry out the substances
 a metabolic waste product from the  Purpose – collect urine that it is in the
breakdown of protein and amino acids in the bladder
liver 2. Catheterization – done by inserting catheter
 50% of the formed elements (solid particles)  inserting tube that will go in bladder and it
is urea will be able to collect clean urine
 Creatinine and Uric Acid – used as marker for 3. Supra-pubic aspiration – insert a needle
urine above the pubic where the bladder is located
 a fluid can be identified as urine if it contains
a high concentration of urea and creatinine QUALITY CONTROL MEASURES
In order to maintain good quality:
 Chloride – major inorganic solid dissolved in
1. Must be analyzed within 1 hour from
urine, followed by sodium and potassium.
collection
 A small amount of protein, mainly albumin, is
 beyond 1 hour you will ready expect
excreted in urine and urobilinogen is typically
deterioration of crystals and increase in
present.
bacteria
 In urinary sediment, a few squamous,
2. Refrigerate at 2 – 8oC for not more than 8
transitional, and renal epithelial cells per
hours
highpower field (40X) as well as one to two red
 in cases wherein you cannot perform the
blood cells (RBCs) or one to five white blood
test within 1 hour
cells (WBCs) are considered normal findings.
3. Preservatives (boric acid) may be used
 Mucus and one to two hyaline casts per low-
 preservation of urine only happens (rarely)
power field are common.
in extreme cases
 Sperm cells are occasionally observed in a
urine specimen from a female, but not a male.
 Amorphous urate and phosphate crystals, ROUTINE URINALYSIS
calcium oxalate, triple phosphate, and uric 1. Physical Examination
acid crystals are common findings in urine The table below indicates the Normal physical
sediment. attributes of urine
NORMAL
pigments: urochrome,
color straw to amber
uroblin, uroeythrin
odor volatile acids aromatic
related to pH and
turbidity clear
specific gravity
750-2000
volume depends on diet
ml/24hrs
specific
over density of water 1.005-1.030
gravity
pH acidity or alkalinity 4.5-8.0

 COLOR: straw (colorless or tinge of yellow) to

SPECIMEN COLLECTION
1. Early morning urine – preferred specimen
because it is concentrated from overnight
retention in the bladder
 most accepted urine sample for lab testing
amber (pearl-like in color or dark yellow)
 ODOR: fecaloid in odor is indication of bacterial
infection
 TURBIDITY: if urine is turbid, there are particles
that are abnormally increase values
 VOLUME: 2L everyday
 SPECIFIC GRAVITY: if it goes beyond 1.030
the urine is turbid
 pH: (<) aciduria (>) alkaline
2. Microscopic Examination
 Centrifuged urine is decanted and the
sediments are placed on a clean glass slide for
microscopic examination.
 Decanting – simply turning test tube upside
down (because of the capillary action of test
tube, liquid pour out and sediments stay)

 We use both the low power objective as well as HISTOPATHOLOGY

the high power objective to quantitate
microscopic structures while others are graded  histo – study of tissue, pathology – study of
qualitatively. disease (history, pathogenesis, treatment)
 The work of the Medical Laboratory Scientist is
The following are the possible microscopic basically preparing the tissue sample for the
structures seen in pathologic urine. pathologist to view under the microscope.
 The quality of the tissue sample to be studied
Term relies on our skills in the preparation of the
Cellular tissue sample.
Elements
RBC hematuria
The following are the steps involved in tissue
WBC (pus cells) pyuria
Epithelial cells
processing:
cylinduria or 1. Fixation – submerging in 10% formalin
Casts  to keep the tissues together that prevents it
cylindroiduria
from shredding and falling apart
crystalluria (renal 2. Dehydration – submerging in three increasing
Crystals
stones) concentration of ethyl alcohol (40%-70%-95%)
Description  increase in concentration will remove water
Acidic Calcium oxalate envelope shape slowly, so it will not damage the tissue
Amorphous urates sand grain-like sample
Uric Acid polymorphic  removing the water component in the tissue
rectangular plates sample (water will cause changes in tissue)
Cholesterol with notched
corners
3. Clearing – aka: de-alcoholization; uses xylene
Cystine flat hexagon  removing the alcohol by submerging to
xylene (toxic, cancerous, volatile)
Alkaline Calcium carbonate dumbbell shape 4. Infiltration – filling up of tissue spaces with
Amorphous paraffin wax
sand grain-like
Phosphate  at 58-60 degrees temperature
Triple phosphate coffin lid 5. Embedding or Molding – putting the tissue
Calcium colorless thin into cassettes
phosphate prisms or threads  putting again paraffin wax to solidify
Ammonium  cassettes (1in x 1in) so it will give ease to
thorny apples
buirate
trimming
Others spermatozoa not reported
6. Trimming – removal of excess paraffin
yeast cells budding RBC-like 7. Sectioning – aka: microtomy; uses microtome
Trichomonas to cut blocks into ribbons at 5 um (micrometers)
parasites  tissue sample submerged in hot water bath
vaginalis
bacteria at 45 degrees Celsius and insert the glass
thread-like slide
mucus threads
structures  remove the paraffin by heating in a Bunsen
burner
8. Staining – uses H&E or hematoxylin and eosin
 hematoxylin (purple) – basic dye, stains the
acidic components of tissue
 eosin (orange/red) – acidic dye, stains the
basic components of tissue
9. Mounting – cover slip with mounting fluid
“Eukitt” about 1 or 2 drops depending on the
size of cover slip
 cover with cover slip to preserve and protect
 alternative to “Eukitt” is clear nail polish
10. Labeling – for identification of the slide
 slides are with frosted ends (unclear
portion) to place the label

AND NOW READY FOR ANATOMIC

PATHOLOGIST TO CHECK WHETER NORMAL
OR ABNORMAL.