Sylvia Rahmayati, dr.

, SpPK
+Definition

+The purpose of lab examination



+ Steps of lab examination:- Preanalytic


- Analytic


- Postanalytic
+Laboratory samples:- Urine
- Blood
- Faeces
- CSF
- Transudat&exudat
- Sperm
- Et all
CLIN
PATH
I. ROUTINE
CLIN LAB
EXAM
II. SIMPLE
CLIN LAB
EXAM
A. DEFINITION OF ROUTINE CLIN. LAB
B. EXAM.
C. TO ESTIMATE A LIMITATION
BLOOD.
URINE.
FAECES.
Hb.
LEUKO. CONT.
DIFF. COUNT.
B.S.R.
MACR/MICR
PROT/RED.
MACR/MICR.
III. MANAGEMENT OF ROUTINE & SIMPLE CLIN. LAB
A. EXAM
B. TO ESTIMATE A LIMITATION
BLOOD.
URINE.
FAECES.
S ERY,THROMBO,
RETIC
PCV, MCV, MCHC
P. SLIDE/PARAS
BLOOD GROUP
RL/BT/CT/CR
SPUTUM.
CEREBRO SPINAL FLUID
TRANSUDATE / EXUDATE
SECRETE : VAG. / UR. / INJ.
CEMEN ANALYSIS.
UROB.
BIL
BENZ
ESBACH
PREG T.
SED
ACID
SP. GR
URINE SPECIMENS

1. THE MORNINGS URINE
THE URINE IS CONCENTRATED
THE FIRST VOIDED
BEST FOR EXAM : SEDIMENT, S.G.,
PREGNANT TEST
2. AT THE TIME URINE
3. TWO HOURS POST PRANDIAL URINE
FOR EXAMINATION OF GLUKOSE
4. 24 HOURS URINE
PATIENT
GETS UP
URINATES
(AT 06.00)

DISCARDED
URINE PASSED DURING
THE REST OF THE DAY
AT 06.00
THE OTHER DAY
URINE
(MORNING)
(NEW)
SHAKE
MACROSCOPIC

COLOUR
SMELL
CLOUDY
ACIDITY
SPEC. GRAF
SEDIMENT
MICROSCOPIC

ERYTHROCYTE
LEUKOCYTE
EPHITEL
CRYSTAL
CAST
CHEMIC

ALBUMIN
GLUCOSE
UROBILIN
BILIRUBIN
KETOBODY
BENZIDIN
ROUTINE
SIMPLE
SUPERNATANT
LIGHT YELLOW (TEA) NORMAL
DARK YELLOW BILIRUBIN (?)


FOAM TEST
SHAKE
(HARDLY)
FOAM
YELLOW (OBVIOUS)
= F. T +
> BIL. +

DUBIOUS


FOUCHET
RED (BLOOD ?)

SED. EXAM ERYTHROCYT : (+) = HEMATURI
(-) = Hb. UR

BENZIDIN TEST
THE OTHER COLOUR
+FOOD / VEGETABLES GREEN
+DRUGS : ANTIPIRIN YELLOW
FENACETIN
+SUBST. FENOL, SALICYL DARK GREEN
A. COLOUR
1. MACROSCOPIC EXAMINATION OF URINE
B. TURBIDITY (NORMAL : CLEAR)

REDDISH BLEEDING SEDIMENT ?
(ERYTHROCYT)
SMOOTH (WHITE) BACTERIA (GRAM)
DENCE (WHITE) (ALKALIC / NEUTRAL URINE)
- PUS
- PHOSPHATE / CARBONATEE CRYSTALS

+ ACETIC ACID SOL (6%)

REDUCE / DISAPPEARED

SPERMATOZOA

VOLUME OF URINE NORMAL : 800 1600 ml/24 H
2
S
4 DAY 3X NIGHT
+POLYURIA D.M., D.I., CHR. NEPHRATIS, EDEMA,
RECONV. FROM CHR. DISEASES

+OLYGURIA ACUTE NEPHRITIS, ECLAMPSIA, ENTERITIS,
HEAVY PERSPIRATION,
DECOMP. CORDIS.

+ANURIA COLLAPS, Hg CL
2
INTOXICATION
C. ACIDITY (pH) (N. 4.7 - 7.5) AVER. 6.0

LITMUS PAPER

BLUE RED = ACID
BLUE = ALKALINE
RED VIOLET = NEUTRAL
MUST BE DONE ALWAYS :
- ALBUMIN TEST ACID URINE
- INTERPRETATION : MORE EASY

ADV. : 1. NEW URINE ALKALINE UTI = UREA SPL.M.O
2. GUIDE OF ACIDOSIS TH/ BY ALKALIN SUBST.
D. SMELL
NORMAL URINE SMELLING
ABNORMAL JENGKOL SMELLING

JENGKOL INTOXICATION

+ ALBUMINURIA
HEMATURIA
CRYSTALURIA

FRUITS KETONURIA
AMONIAK UREUM OF BACTERIA
E. EXAMINATION OF SPECIFIC GRAVITY (S.G.)
NORMAL : 1.010 - 1.025 (1.020)
LOW S.G. ( < 1.010 ) = KIDNEY OR ENDOCRINE DISORDER
HIGH S.G. ( > 1.025) = NEPHR.DEG. / FEVER GLYCOSURIA
METHOD & EQUIPMENT
= URINOMETER
= MEASURING CYLIDER (50 ml)
+ TEMP. : EVERY 3
0
C > 15
0
C : + 0.001
4
0
C > 17
0
C : + 0.001
+ GLUCOSE : EVERY 270 mg/DL : -0.001
1 % : -0.004
+ PROTEIN : EVERY 400 mg/DL : -0.001
1% : -0.003
IF THE AMOUNT OF URINE IS SMALL
USE : - FALLING DROP METHOD
- REFRACTOMETER
S.G. IS DEPEND ON THE TOTAL OF SOLUTE SUBSTANCES
1.000
1.020 CORECTION
1.040
CLINICAL VALUE OF URINE SPECIFIC GRAVITY.
TO HELP THE DIAGNOSIS GLUCOSURI OF COMATOUS PATIENT
FISHBERG CONCENTRATION TEST FOR RENAL FUNCTION
PATIENT DONT DRINK WATER / EAT WATERY FOOD
AT 18.00 BUT MAY URINATE AT THE TIME
IN THE MORNING TAKE MORNING URINE


EXAM. SPECIFIC GRAVITY

S.G. 1.020 GOOD

1.020 - INTERUPTED RENAL FUNCTION
= GNA
- NIGHT EDEMA
= DECOMP. CORDIS
ADVANTAGE :
EASY - DO NOT GIVE TROUBLE TO PATIENT
TO KNOW RENAL FUNCTION
GET URINE WITH S.G. HEIGHT FOR EXAMINATION
SUBSTANCES WHICH DIFFICULT TO BE DETECTED
IN HEIGHT SPECIFIC GRAVITY URINE
METHODS
2. MICROSCOPIC EXAMINATION OF URINE
NEW URINE < 6 HOURS
CENTRIFUGE AT 1500 RPM / 5 MINUTES
SEDIMENT
COVER WITH
COVER GLASS
SLIDE
MICROSCOPE OBJECTIVE 40 X
EYEPIECE 10 X
CONDENSOR
EXAMINATION ! !
CRYSTAL
ERITHROCYTE / LOW POWER
LEUKOCYTE / HIGH POWER
C A S T / LOW POWER
EPHITEL CELL
ORGANIC
SEDIMENT
ANORGANIC
SEDIMENT
ERITHROCYTE
MORPHOLOGIC :
A. NORMAL (NEW URINE) :
- ROUND, F + 7 m (EQUALLY)
- YELLOWISH

B. CRENATED (HIGH SPECIFIC GRAVITY URINE)
- DARKER AT THE EDGES
- SPIKY EDGES
- THE FLUID OUT OF THE CELLS

CLINICAL VALUE :
ERY + URINE : - BLEEDING (CANCER OF REN, PYELUM)
(HEMATURI) - TRAUMA (CALCULUS, CRYSTAL)
- INFLAMMATION (TBC, GNA)
FAR ADVANCED EXAMINATION
NORMALLY, THERE ARE NO RED CELLS IN URINE
NOTES :
ERY + : : REPEATED BY MIDSTREAM URINE
OR CATHETER URINE
NORMAL : 0 - 1 / LPB
LEUCOCYT (WHITE CELLS)
MORPHOLOGIC :
CLEAR GRANULAR DISKS
ROUND ; F + 11 m (1.5 - 2 E)
THE EDGES NOT CLEAR
CELLS SURFACES ARE GRANULAR
IN NEW URINE AMOEBOID MOVEMENT
IN NEW ALKALIST CLUMPS
IN ALWAYS FOUND 2 - 6 LEUCOCYT OR MORE / LOW POWER
- CATHETER URINE
- MEN
> 6 / HIGH POWER = PATOLOGIC
THE CLINICAL VALUE : N : 0 - 6 / LOW POWER
E
E
NOTES :
CLEAN VOIDED URINE (GEWASSEN URINE)
: - OPEN PREPUTIUM
- CLEAN URETHRAE
- TAKE MIDSTREAM URINE
: - WASH THE AREA AROUND URETHRAE
- OPEN LABIA
MIDSTREAM URINE
CATHETERIZATION
DANGEROUS, INFECTION
EPITHELIAL CELLS
MORPHOLOGIC :
SCUAMOUS
EPITHELIAL CELLS
(VAGINA)
(URETHRAE DIST.)
ROUND EPITHELIAL
(TUBULUS)
CAUDATUS
EPITHEALIAL CELLS
(PELVIS R.)
SMALL AMOUNT OF EPITHELIAL : USUALLY,
(ESPECIALLY ON WOMAN)
DIAGNOSTIC VALUE IS SMALL
CRYSTAL
IN ACID URINE : URIC ACID URATE
URATE CRYSTAL / AMMORPHUS URATE
Ca OXALATE
IN ALKALIST URINE : AMMORPHUS MAGN. PHOSPHAT
Ca PHOSPHAT / CARBONATE
AMMORPHUS PHOSPHAT
AMMONIUM URATE
URIC ACID IN FRESH URINE CALCULUS IN THE U.G.
OTHERS, THERE HAVE NO CLINICAL VALUE
- Ca OXALATE :
SIZE VARIABLE
CLEAR
MORPH. LIKE ENVELOPE
- PHOSPHAT :
TRIPLE
PHOSPHAT
CALCIUM
PHOSPHAT
AMMORPHUS
PHOSPHAT
CLEAR
- CARBONATE :
CALSIUM
CARBONATE
- URIC ACID :
- URATE :
NH
4
URATE
Na - URATE
AMMORPHUS
URATE
COLOUR
IS BROWN
APPEARENCE OF CRYSTAL
CRYSTAL
NORMAL CRYSTALINE DEPOSITE
1. CALCIUM OXALATE (ACID URINE)
A. SHAPE LIKE ENVELOPE
SIZE 10 - 20 m m
B. SHAPE LIKE PEANUTS
SIZE + 50 m m
COLOUR : CLEAR / TRANSPARANT
( COLOURLESS )
2. URIC ACID (ACID URINE)
SHAPE VARIES (SQUARE, DIAMOND SHAPE, CUBICAL/
ROSE SHAPE)
SIZE 30 - 150 m m
COLOUR YELLOW - BROWNISH RED
3. TRIPLE PHOSPHATES ( NEUTRAL / ALKALINE URINE )
SHAPE : A. RECTANGULAR
B. LIKE A TERM LEAF / STAR
SIZE 30 - 150 m m
COLOUR : COLOURLESS
4. URATES (ALKALINE / CLEAR)
SHAPE LIKE : 1. CACTUS
2. A BUNDEL OF NEEDLE
5. LESS COMMON CRYSTAL
A. CALCIUM PHOSPHATE (ALKALINE NEUTRAL)
B. CALCIUM CARBONATEE (ALKALINE NEUTRAL)
C. CALCIUM SULFAT (ACID URINE)
CAST
CAST OF SEDIMENT IS PRECIPITATE OF PROTEIN
IN TUBULI IN ACID URINE
CYLINDRICAL IN SHAPE AND LONG
PROTEIN
1. HYALINE CASTS :
TRASPARENT, SLIGHTLY RE-
FRAQTIL, THE END RONDED
OR TAPERED
2. GRANULAR CASTS (COARSE) :
RATHER SHORT CASTS FILLED
WITH LARGE GRANULES PALLET
PALE YELLOW IN COLOUR
(GRANULES COME FROM DE-
GENERATE EPHITELIAL CELLS FROM
THE TUBULES OF THE KIDNEY)
3. FINE GRANULAR CAST :
GRANULES ARE SMALLER
AND DO NOT FILL THE CAST
4. BLOOD CASTS (ERITHROCYTES CASTS) :
CASTS FILLED WITH RED BLOOD
CELLS BROWNISH IN COLOUR
5. PUS (LEUKOCYTES) CASTS :
CAST FILLED WITH LEUKOCYTES
6. EPHITELIAL CASTS :
CAST FILLED WITH PALE YELLOW
EPHITELIAL CELLS
BIOCHEMISTRY EXAMINATION OF URINE
1. PROTEIN EXAMINATION OF URINE
> PRINCIPLE : PROTEIN CLOUDY

> CONDITION OF URINE : - ACID AND CLEAR
> CHARACTER OF EXAMINATION : QUALITATIVE /
SENSITIVITY 5 - 10 mg /dL
CERTAIN
pH
A. EXTON TEST FOR PROTEIN URINE
REAGEN : SULFOSALISILIC ACID : 25 GR. 50
NA2SO
4
: 100 GR. 88
AQUADEST : 500 CC. 1000
URINE


SENTRIFUS
SHAKE
REAGENS : 2.5 ml
URINE : 2.5 ml
EXTON + : - PROTEIN
- PROTEOSEN
- BENCE JONES PROTEIN
- URIC ACID & OTHERS
CLEAR = PROTEIN -
(NO WHITE PRECIPITATE)
TURBID = EXTON +

FOLLOWED BY :
1. BANG TEST
2. ACETO PRECIPITABLE
SUBSTANCE TEST
COMPARE WITH A TUBE OF UNTREATED URINE
AGAINTS A BLACK BACKROUND
B. BANG SEMI QUANTITATIVE TEST FOR PROTEIN URINE
1) REAGEN :
SODIUM ACETATE : 11.8 GR
ACETIC ACID GLASIALE : 5.65 CC
AQUADEST ad 100 CC
2) METHODS
IF THE AMOUNT OF ALBUMIN 3000 mg % CLOTTING
BOILED
EXTON
PROTEIN :
BANG
PROTEOSEN
PROTEIN
BENCE JONES PROT.
TURBID WITHOUT BOILING
ACETOPRECIPITABLE
RESULT :
SYM PROT
BOL (mg %)
CLEAR - 0
SLIGHT TURBID + + 10
TURBID WITHOUT + 10 - 50
GRANULE
TURBID WITH ++ 50 - 200
GRANULE
TURBID WITH +++ 200 - 500
FRAGMENT
CLOTTING ++++ > 500
REAGEN 0.5 ml
URINE 5 ml
100
0
C


WATER BATCH
BOILED
10
READ
+
++
+++
-
-
+
-
+
C. QUANTITATIVE PROTEIN URINE EXAMINATION
(ESBACH)
- TO CONFIRM QUANTITY OF PROTEIN IN URINE
- TO BE DONE FOR : 24 HOURS URINE

REAGENT : > PICRIC ACID : 1 GR
> CITRIC ACID : 2 GR
> AQUADEST : 100 CC
METHOD OF THE TEST (ESBACH)
PIPET URINE
PIPET REAGEN
PUT THE RUBBER OR CORKY SHUTTER
SHAKE
MOVING
THE TUBE
(10 X)
PLACE :
- IN ROOM TEMP.
- FOR 24 HOURS
- AVOID TO LIGHT AND
UPRIGHT POSITION
ALBUMINOMETER
READ :
THE HEIGHT
OF WHITE
PRECIPITATION
5
5 GR / L / 24 HOURS
TEST FOR ACETOPRECIPITABLE SUBSTANCE
+ WATER FULL
URINE 10 ml
30% ACETIC ACID
SOLUTION
0 gtt
Urine
2 ml
Urine
2 ml
Urine
2 ml
I II III
CLEAR TURBID
RESULT : SECOND TUBE : - CLEAR : ACETOPRECIPITABLE
- TURBID : MUCIN
E. EXAMINATION OF BENCE JONES PROTEIN
CHARACTERISTIC OF BENCE JONES EXAM. :
- PRECIPITATE : 44
0
- 50
0
C
- SOLUBLE AT TEMPERATURE > 95
0
C
SCREENING TEST :
URINE
5 ml
HCL CONCENTRATE
ADD HCL CONCENTRATE GENTLY,
SO THAT LIMITATION BETWEEN
URINE AND HCL
PRECIPITATION
RING
= B.J.P.
= - GLOBULIN
- PROTEOSA
- B - S - P
CONFIRMATION TEST :
10 ml
URINE
1 DROP ACETIC ACID SOLUTE 2%
pH : 5.5
ADD Na Cl CONCENTRATE 2 ml
READ THE TEMPERATUR
IN WHICH ITS PRECIPITATION
OCCUR
: - MULTIPLE MYELOMA
- CLL, CML
- AMYLOID DESEASE
- -
+
+
100
0
C


WATER BATCH
2. DETECTION OF GLUCOSE IN URINE
PRINCIPLE : GLUCOSE IS A REDUCING SUBSTANCE IT REDUCES THE BLUE
COPPER SULFATE OF BENEDICT SOLUTION TO RED COPPER
OXYDE WHICH IS INSOLUBLE.
A. BENEDICT METHOD
- REAGENT :
TRISODIUM CITRATE 85 GR 173
SODIUM CARBONATE 135 GR. 100
DISTILLED WATER 350 GR.

COPPER SULFATE 85 GR 17.3
WATER 50 ml 100
MIX AND ADD
500 ml WATER
- METHOD :
BENEDICT 5 ml
URINE 8 GTT
BOIL
100
0
C
WATER BATCH
5
LEAD TO COOL/
ROOM TEMP.,
READ THE RESULT
SIGN GLUCOSE
gr/dl


BLUE - 14
0-0.1



TRACE GREEN + 28
WITH YELLOW




YELLOW ++ 56





BROWN +++ 83





ORANGE ++++ 111
TO BRICK RED
A. DIABETES
MELLITUS
B. SECONDARY
GLUCOSURIA
C. GLUCOSURIA
RENAL
ALIMENTARY
HYPERTHYROID
CUSHING
SYNDR.
ACROMEGALIA
INTRACRANIAL
HYPERTENTION
A. LACTOSE
B. FRUCTOSE
C. PENTOSE
D. GALACTOSE
SALICYL
ANTIPYRIN
UROTROPIN
SANTONIN
URIC ACID
KREATININ
INDIKAN
REDUCTION
+
A. SUGAR
B. THE OTHER
SUBSTANCES
GLUCOSE
THE OTHER
SUGAR
DRUGS
THE OTHER
SUBSTANCES
BILIRUBIN METABOLISM
ERI
HAEMOGLOBINE
HAEM + GLOBIN + Fe
BILIVERDINE
BILIRUBIN
PREHEPATIC BILIRUBIN
INDIRECT BILIRUBIN
- WATER IN SOLUBLE
- ALCOHOL SOLUBLE
STERCOBILINOGEN
CONJUGATION
WITH GLUCURO-
NICACID
CONJUGATED
BIL.
UROBILINOGEN
UROBILIN
RUPTURE
FAECES
(STERCOBILIN)
DIRECT BILIRUBIN
- WATER SOLUBLE
REN
DETECTION OF UROBILINOGEN AND UROBILIN IN URINE
A. UROBILINOGEN (FRESH URINE + Na CARBONATE)
= WALLACE DIAMOND
PRINCIPLE : UROBILINOGEN + PARADIMETYL BENZALDEHYDE
COMPOUND WITH WINE RED COLOUR (CHERRY)
METHOD : EHRLICHS REAGENT
R/ : - P. DIMETYL AMINO : 2 gr
BENZALDEHYDE
- 37% HCl : 50 ml
- AQUADEST : 100 ml
REAGENT 10 ml
URINE 5 ml
SHAKE
+ DEEP RED COLOUR
- A FINE PINK COLOUR
NORMAL : 2 mg / 24 HOURS
B. DETECTION OF UROBILIN IN URINE
DISTURBANCE WITH EHRLICH
PRINCIPLE : UROBILINOGEN UROBILIN
+
Zn - ACETAT - ALC.
COMPOUND OF Zn. UROBILIN
(GREEN FLUORESCENS)
MATERIALS : 1. LUGOL IODINE SOLUTION
R/ IOD 0.5 gr + WATER 150 ml
IOD KALI 1 gr
2. SCHLESSINGER REAGENT
R/ Zn - ACETAT : 10 gr
ALKOHOL 96% : 100 ml
OXYD.
IOD
7.5 ml REAGENT
2 GTT LUGOL
5 ml URINE
SHAKE
A PRECIPITATE FORMS
STRONG GREEN FLUORESCEN = +
LIGHT GREEN FLUORESCEN = NORMAL
UROBILINURI :
- HEMOLISES = A.N HEMOLITICS
- HEPATITIS ETC.
METHOD :
DETECTION OF BILIRUBIN IN URINE
IF THE COLOUR OF URINE IS YELLOWISH BROWN FOAM TEST
1. FOAM IS YELLOW BILIRUBIN +
2. FOAM IS NOT YELLOW BILIRUBIN -
3. UNCERTAIN FOUCHET TEST
A. TEST USING FOUCHET REAGENT
MATERIALS :
1. FOUCHETS SOLUTION
R/ TCA 25 gr
10% FeCl
3
10 ml
AQUADEST 100 ml
2. 10% SOLUTION OF CACl
3

3. SOLUTION OF Na
2
CO
3
CONCENTRATE
PRINCIPLE : BILIRUBIN BILIVERDIN (GREEN)
OKS
FeCl
3

METHOD :
Na2CO
3
20 GTT
CaCl
2
10 GTT
U 5 ml
SHAKE
A PRECIPITATE FORMS
: COLOUR : GREEN BLUE
RESULTS : - = NO COLOUR CHANGE
+ = THE PRECIPITATE TURNS GREEN / BLUE
+
B. HARRISON/HAWKINSON MODIFICATION
TO DETECT BILIRUBIN IN URINE
RINSE
LARGE
FILTER PAPER
10% BaCl2 SOLUTION
ALLOWED TO DRY
AND MAKE LITTLE PIECE OF
FILTER PAPER BY CUTTING
I. URINE
II. FOUCHET SOLUTION
TEST :
1. PUT ONE DROP OF URINE
2. PUT ONE DROP OF FOUCHET SOL.
GREEN TO BLUE COLOUR
NOTES :
1. FOUCHET TEST :
- VERY SENSITIVE
- DIAGNOSIS CAN MADE EARLY
2. BILIRUBIN + BY :
A. OBSTRUCTION OF DUCTUS BILIARIS :
- CALCULUS
- INFLAMATION
- TUMOR, ETC
B. HEPATITIS
TEST
+ =
DETECTION OF KETONE SUBSTANCE IN URINE
KETONE SUBSTANCE : CONSIST OF :
1. ACETON : 2%
2. ASETOASETIC ACID : 20%
3. BETAHYDROXYBUTYRIC ACID : 78%
AS A RESULT OF FAT METABOLISM
IF ALLOWED TO STAND FOR A WHILE:
B.HYDROXYBUTYRIC A. ACETOACETIC A. ACETON
MUST BE USED FRESH URINE
INDICATION FOR KETONE SUBSTANCE DETECTION IS
REDUCTION TEST ( +++ / ++++)

URINE : REDUCTION +++ / ++++
KETON SUBSTANCE ( PRECOMA)
THERE IS NO SPECIFIC TEST FOR B.HYDROXYBUTYRIC A.
CLINICALLY :
- ACETON +
- ACETOACETIC ACID + DISTURBANCE OF METABOLISM
+
METHODS :
1. ROTHERA METHOD
2. GERHARDT METHOD
KETOSIS DIABETICUM
A. ROTHERA METHOD
PRINCIPLE :
- ACETON
- ACETOACETIC ACID
+
Na. NITROPRUSIDA
SUBSTANCE WITH
VIOLET COLOUR
MATERIAL : A. Na. NITROPRUSIDA 5 gr
AMMONIUM SULFATE 200 gr
B. AMMONIUM HYDROXIDE CONCENTRATE


METHOD:
MAKE COMPLETE
SOLUTION
URINE 5 ml
REAGENT 1 gr
NH4OH
CONCENTRATE
READ RESULT
WITHIN 5
VIOLET COLOUR RING
+ BRIGHT VIOLET
++ DARK VIOLET
+++ DARK AND WIDE
VIOLET
SENSITIVITY :
ACETON = 1 : 20.000 (+) : 1 - 5 mg%
ASETOAACETIC ACID = 1 : 400.000 (+) : 10 - 25 mg%
B. HYDROXYBUTYRIC A.= 0
ROTHERA : - VERY SENSITIVE
- + IN PERSON AT STARVATION OR FASTING
CONDITION

B. GERHARDT METHOD
FOR ASETOACETIC ACID ONLY
PRINCIPLE :
ASETOACETIC ACID + FeCl
3
WINE RED SUBST.
THIS TEST IS LESS SENSITIVE THAN ROTHERA METHOD
REAGENT : 10% FeCl
3
SOLUTION
METHOD :
2 DROPS
FeCl
3
SOLUTION
URINE 5 ml
SHAKE
+
(RED)
ASETOACETIC ACID

FALS POSITIVE :
- FENOL
- SALICYLAT
- ANTIPYRIN
- Na. CARBONATE
MUST BE DONE
ROTHERA
METHODS
OR
SECOND GERHARDT
METHODS
AQ. 5 ml
FeCl3
U : 5 ml
CHANGE TO - :
KETONE SUBSTANCE
STILL + :
THE OTHER SUBSTANCES
U : 5 ml
COOK
DETECTION OF OCCULT BLOOD IN URINE
IF : HEMATURIA SEDIMENT EXAM. ERI +
Hb. URIA SEDIMENT EXAM ERI -
BENZIDINE TEST
PRINCIPLE :
H
2
O
2
H
2
O + On BENZIDINE OXIDATION
METHOD :
1. BENZIDINE
SOLUTION
5ml ACETIC ACID
GLACIAL
1 gr BENZIDINE
2. URINE
COOK
TEST
URINE
MAKE COOL
READ WITHIN
5
3. H202 SOLUTION
THIS TEST IS VERY SENSITIVE
GIVE + RESULT BY OXYDASE
FROM LUEKOCYTE
- URINE MUST BE HEATED
- EQUIPMENT CLEAN
BLOOD PEROKSIDASE ACTIVITY
PREPARE :
URINE TEST STRIP
CHARACTERISTIC OF THE TEST :
RAPID, EASY, SPECIFIC AND CHEAP
MATERIALS :
TEST STRIP
SPECIFIC GRAVITY
NITRITE
pH
PROTEIN
GLUCOSE
KETOBODY
UROBILINOGEN
BILIRUBIN
BLOOD
PLASTIK ROD
NYLON COVER
TEST FIELD
(PAPER CONTAIN REAGENT)
FILTER PAPER
PROCEDUR OF THE TEST :

1. IMMERSE THE TEST STRIP
FOR APPROX. 1 SECOND
2. REMOVE EXCESS URINE FROM THE STRIP
BY WIPING THE EDGE OF URINE ON
THE CONTAINER (TUBE)
URINE
READ :
COMPARE
THE COLOUR CHART
UROTRON
PREGNANCY TEST
PRINCIPLE : - THE PREGNANT WOMEN URINE CONTAIN HCG
( HORMON CHORIONIC GONADOTROPIN)
- THIS TEST IS BASED ON ITS ABILITY TO SHOW
AN EXISTANCY HCG IN THE WOMEN URINE
BY IMMUNO CHEMICAL REACTION

HCG + ANTI-HCG ANTIGEN ANTIBODY REACTION

ANTIGEN = HCG
ANTIBODY = RABBIT ANTI SERA (ANTI-HCG)
INDICATOR SYSTEM =
ERYTHROCYTE COATED BY HCG HEMAGLUTINATION
LATEX COATED BY HCG LATEX AGLUTINATION
1 MINUTE
METHOD :

1. DROP 1 GTT URINE
2. DROP 1 GTT ANTI-HCG

MIXED ( + 1)

3. DROP 1 GTT LATEX

MIXED (MOVED SLIDE GENTLY)
READ : AGGL - PREG TEST +

AGGL + PREG TEST -
HEMATOLOGY
HEMATOLOGI IS THE STUDY OF BLOOD INCLUDES THE BLOOD
CELLS AND FLUID
VOLUME OF BLOOD : ADULT WITH 60 KG + 6 LITERS
BLOOD COMPOSITION :
BLOOD
BLOOD FLUID
(55%)
BLOOD CELLS
(45%)
ERYTHROCYTES
LUEKOCYTES
THROMBOCYTES
(PLATELETS)
WATER
(90%)
SUBSTANCES
(10%)
PROTEIN :
- ALBUMIN
- GLOBULIN
- FIBRINOGEN
CARBOHYDRATE
LIPID
VITAMIN
MINERAL
ENZYM
BLOOD COLLECTING PROCEDURES
I. CAPILLARY BLOOD
A. LOCATION :
ADULT : THE TIPS OF FINGER
BABY : HEEL OR BIG TOE
WITHOUT : - CYANOTIC
- HAEMORRHAGIC
- PALE

B. METHOD :
CLEAN THE SITE
1. WITH THE COTTON SWAB
+ 70% ETHANOL
2. WITH DRY COTTON SWAB
PRICK THE SITE FIRMLY AND
RAPIDLY WITH DISPOSIBLE
BLOOD LANCET (AUTO CLICK)
DEEP ENOUGH ( 2 mm DEPTH)
TO GET GOOD BLOOD DROPS
WIPE AWAY THE 1
st
DROP OF
BLOOD WITH DRY COTTON
WOOL
X
3
4
COLLECTION OF VENOUS BLOOD
PRINCIPLE : VENOUS BLOOD IS COLLECTED FROM A VEIN
IN THE ARM WITH A NEEDLE AND SYRINGE
MATERIALS :
1. FOR DISINFECTING SKIN :
- 70 % ETHANOL
- COTTON WOOL

2. FOR VENEPUNCTURE
- TOURNIQUET
- NEEDLE (STERIL)
VARY IN LENGTH / DIAMETER

3. FOR COLLECTION OF BLOOD
- SYRINGES ( OF 2, 5, 10 OR 20 ml)

4. BOTTLES OR TUBES
- EMPTY = CLEAN / DRY
- CONTAIN ANTI COAGULANT
(CRYSTAL / SOLUTION)
METHOD :
READ CAREFULLY THE PATIENTS REQUEST FOR
ESTIMATE HOW MUCH THE BLOOD IS NEEDED
BEFORE TAKING THE BLOOD
WASH YOUR HAND WITH SOAP AND WATER
POSITION OF PATIENTS
- IN LABORATORY
- IN BED
TORNIQUET
& NEEDLE
A.C A.C
THE CORRECT SITE TO TAKE VENOUS BLOOD
IS THE VEIN IN THE BEND OF THE ELBOW
POINT 1, 2, 3, AND 4
APPLY THE TOURNIQUET :
1. WRAP THE TOURNIQUET FIRMLY AROUND THE ARM
2. PULL ONE OF THE END ACROSS
3. LOOP IT UNDER THE MAIN PART OF TOURNIQUET
- JUST TIGHT ENOUGH SLOW DOWN THE BLOOD FLOW
- DISTEND THE VEIN
IT MUST THE BLOOD FLOW IN
NOT BE SO TIGHT ARTERIES DIMINISHED
4. IN ORDER TO SWELL THE VEIN :
ASK THE PATIENT TO CLOSE AND OPEN HIS HAND SEVERAL
TIMES
TO GET THE VENOUS BLOOD
1
6
2 7
3 8
4 9
5 10
NOTES :
1. PALPATION TO CERTAIN THE VEIN
2. DESINFECTION OF THE SKIN WITH 70% ETHANOL
3. HOLDING THE NEEDLE / SYRINGE
4. VENE PUNCTURE
- POSITION OF THE NEEDLE WITH THE BEVEL UPPER MOST
- MAKE THE VENE PUNCTURE ENTERING THE VEIN WITHOUT
HESITATION
5. IMPORTANT :
- NEVER APPROACH THE VEIN FROM THE SIDE
- NEVER INTRODUCE A NEEDLE WITH THE BEVEL DOWN WORDS
6. YOU WILL FEEL THE NEEDLE GOING THROUGH :
A. THE SKIN = WHICH IS RESISTANT
B. THE WALL OF THE VEIN = LESS RESISTANT
7. PUSH THE NEEDLE ALONG THE LINE OF THE VEIN TO
A DEPTH OF 1 - 1.5 cm.
8. WITH YOUR LEFT HAND, PULL BACK THE PISTON OF THE
SYRINGE SLOWLY, BLOOD SHOULD APPEAR IN
THE SYRINGE
CONTINUE TO DROW BLOOD TO GET THE AMOUNT OF
BLOOD NEEDED
9. REMOVE THE TOURNIQUET
10. APPLY A DRY SWAB OVER THE HIDDEN POINT OF THE
NEEDLE WITH DRAW THE NEEDLE IN ONE RAPID MOVEMENT
FROM UNDER THE SWAB
ASK THE PATIENT TO PRESS FIRMLY ON COTTON WOOL
SWAB FOR 3 MINUTE, KEEPING HIS ARMS OUT STRETCHED

TO TRANSFER BLOOD FROM THE
SYRINGE TO THE TUBE OR BOTTLE :
REMOVE THE NEEDLE
FLOW THE BLOOD THROUGH THE
WALL OF BOTTLE OR TUBE
TUBE MUST BE :
DRY, CLEAN, FREE FROM DETERGENT SILICON
CLOTTING OF BLOOD
WHEN BLOOD IS COLLECTED IN A GLASS TUBE
A
WITHOUT
ANTICOAGULANT
SOLIDIFIES WITHIN 5 - 20
B
WITH
ANTICOAGULANT
CLOTTING IS PREVENTED
FORMING REMAIN FLUID
THE DIFFERENT :
FIBRINOGEN
-
+
SERUM
BLOOD CLOT
PLASMA
WBC
RBC
THE BLOOD SHOULD BE :

ENOUGH
UNHEMOLYSIS
TO AVOID THE HEMOLYSIS OF BLOOD,THERE ARE SOMETHING
TO DO, LIKE :
- USE CLEAN AND DRY MATERIALS
(SYRINGE, NEEDLE, AND BOTTLE/TUBE)
- DRAW WITH GENTLY, NOT TOO HARD
HEMOLYSIS
SERUM
PLASMA
+ - - +
ANTICOAGULANTS
1. E.D.T.A (ETHYLINE DIAMINE TETRA ACETATE)
POTASIUM OR SODIUM SALT
TO CHANGE Ca
++
NON ION Ca
THERE IS NO INFLUENCE TO SIZE, FORM OF BLOOD CELLS
PREVENT THROMBOCYT AGGREGATION IT IS USE TO
COUNT THROMBOCYT
AVIALABLE SUBSTANCE:
- CRYSTAL = 1 mg EDTA / 1 ml BLOOD
= DIFFICULT TO DISSOLVED
- SOLUTION = 10% SOLUTION
IT IS BETTER TO USE SOON
4
0
C FOR 24 HOURS PCV
THROMBOCYTE
SMEAR BLOOD / EDTA 2 HOURS
SHAKE BLOOD / EDTA 1 - 2
2. HEPARIN
AS ANTI THROMBIN
THERE IS NO INFLUENCE TO BLOOD CELLS
(ESP. LEUKOCYTE/ERYTHROCYTE)
EXPENSIVE, DO NOT USE FOR ROUTINE EXAMINATION
USE : 1 mg HEPARIN / 10 ml BLOOD
3. SODIUM CITRAT 3,8%
AS ISOTONIC SOLUTION
FOR HEMORRHAGIC EXAMINATION
ESPECIALLY : BSR : WESTERGRENS METHOD
EXAMINATION IN HEMATOLOGY
1. HAEMOGLOBIN ESTIMATION
2. LEUKOCYTE COUNT
3. ERYTHROCYTE COUNT
4. DIFFERENTIAL COUNTING OF LEUKOCYTES
5. BLOOD SEDIMENTATION RATE
6. THROMBOCYTES COUNT
7. RETICULOCYTES COUNT
8. P.C.V. (PACKED CELLS VOLUME)
9. THIN BLOOD FILMS
10. BLOOD PARASITES
11. BLOOD GROUPING
12. RUMPLE LEEDS TEST
13. BLEEDING TIME
14. COAGULATION TIME
15. CLOT RETRACTION

HAEMOGLOBIN ESTIMATION
METHODS :
1. TALLQVIST
2. SAHLI
3. PHOTOMETRIC
COLOURIMETRIC
1. TALLQVIST METHOD
MATERIALS : - BLOOD LANCET (DISSPOSIBLE)
- TALLQVIST BOOK
PRINCIPLE :
TO COMPARE THE COLOUR OF BLOOD AGAINTS
THE COLOUR OF SCALE
METHOD :
+ CLEAN AND DISINFECT THE FINGERTIP
+ PRICK WITH HEMO LANCET
+ WIPE THE 1st DROP OF BLOOD WITH DRY COTTON
+ PUT A DROP OF BLOOD ON A PIECE OF FILTER PAPER
FROM THE BOOK
+ ALLOW THE DRY
+ COMPARE THE COLOUR OF BLOOD READ Hb : %
ADVANTAGE : EASY / PRACTICE
DISADVANTAGE :
- RESULT IS NOT EXACTLY
THE COLOUR STANDAR MAY BE CHANGE
BECAUSE ITS MADE FROM PAPER
- IT SHOULD BE CORRECTED BY
THE MORE EXACTLY METHOD
100% 15.8 gr %
2. THE METHOD OF SAHLI
PRINCIPLE : Hb + HCl HEMATINE ACID (COLOUR BROWN)
a
b
THE BLOOD IS DILUTED IN AN ACID SOLUTION
(THE COLOUR OF COMPOUND IS COMPARED BY
COLOUR STANDARD WITH VISUALIZED
MATERIALS :
1. Hb METER SAHLI /HELLIGE CONSIST OF :
Hb PIPET
Hb TUBE
Hb STANDARD
SMALL GLASS ROD
2. HCl 0,1 N
3. AQUADEST
4. DROPPING PIPET
5. HEMOLANCET + COTTON & ETHANOL 70%
METHOD:
1
4
5
6
3
2
1. FILL HCL 0.1 N IN TO THE Hb TUBE UNTIL MARK 2
2. PRICK THE FINGER TIP TO GET SOME BLOOD
3. FILL THE Hb PIPET BY BLOOD UNTIL MARK 20 ( 20 m L)
WIPE THE OUTSIDE OF THE PIPET
4. BLOW THE BLOOD FROM THE PIPET INTO Hb TUBE
AND SOAK IN HCl 0.1 N
RINSE THE PIPET BY DRAWING IN AND BLOWING OUT
THE ACID SOLUTION 3 TIMES
5. MIX BLOOD AND ACID THE MIXURE GIVES A BROWNISH
ALLOW TO STAND FOR 5 MINUTE
6. PLACE THE Hb TUBE INTO THE HAEMOGLOBINOMETER,
STAND FACING THE WINDOW
DILUTE BY ADDING WATER,
STIRING BY SMALL GLASS ROD REMOVE THE ROD
COMPARE THE COLOUR OF THE 2 TUBES
STOP IF THE COLOUR MATCH READ THE MARK REACHED
Hb : g/dl
NOTES :
3. PHOTOMETRIC METHOD
CYANMETHEMOGLOBIN
PRINCIPLE : BLOOD DILUTED
K. CYANIDA (KCN)
K. FERRICYANIDA
DRABKIN DILUTING FLUID:
R/ NaHCO3 : 1.0 gr
KCN : 0.05 gr
K3Fe(CN)6 : 0.20 gr
AQUA : 1000 ml
(EXAM IN SPECTRO)
CYAN-MET Hb
(STABIL )
(Hi CN)

MATERIAL :
SPECTROPHOTOMETER
PIPETS & TUBES
DRABKIN DILUTING FLUID
CYAN MET Hb STANDARD
METHOD :

INTO A TUBE, PIPET :
5 ml DRABKIN DILUTING FLUID
0.02 ml BLOOD
MIX THE CONTENT
OF THE TUBE,
LEAVE FOR 5
READ PHOTOMETRIC (l : 520 nm)
ABSORBANCE OF SAMPLE
ABSORBANCE OF STANDARD
X Hb STANDARD ( 15 g/dL)
NORMAL Hb : ADULT : : 13.5 - 18.0 gr/dL
: 11.5 - 16.9 gr/dL

CHILDREN :
10 - 12 YEARS : 11.5 - 14.8 gr/dL
1 YEARS : 11.0 - 13.8 gr/dL
3 MTH : 9.5 - 12.5 gr/dL
BABY : 13.6 - 19.6 gr/dL
CALCULATION :
MATERIAL :
CONSIST OF
METHODS OF :

LEUKOCYTE AND ERYTHROCYTE
COUNT
FOR COUNTING :
LEUKOCYTE ERYTHROCYTE
1. THOMAS PIPET
2. COUNTING CHAMBER
3. DILUTING FLUID
4. HEMOLANCET
COTTON + 70% ETHANOL
TURK
..
- HAYEM
- FORMOL CITRATE
1
1
COUNTING CHAMBER = IMPROVED NEUBAUER
BURKER , ETC.
A B C
COLOUR BAND
&
NEWTONS RING
CHAMBER DEPTH = 0.1 mm
..
PREPARATION OF COUNTING CHAMBER :
PUT COVER GLASS OVER THE COUNTING CHAMBER
PRESS THE COVER GLASS FIRMLY SO THAT IT IS PROPERLY
ATTACHED, AND COLOURED BANDS CALLED NEWTONS RING
APPEAR BETWEEN THE TWO GLASS SURFACES AND IT MEANS
THAT CHAMBER DEPTH IS 0.1 mm
IT IS CALLED AS : PREPARED COUNTING CHAMBER
COUNTING CHAMBER OF BURKER
..
1 mm 1 mm
0.05 mm
1 2 3
X
= LEUKOCYTE COUNT SQUARES
1
5
1
5
1
10
X X mm
3

= 25 SQUARES
A
= ERYTHROCYTE COUNT SQUARES
1
20
1
20
1
10
X X mm
3

= 80 SQUARES
B
=
1
5
1
20
1
10
X X mm
3

= 20 SQUARES
VOL =
1
20
1
20
1
10
X X
mm
3
= N
80 X
1
50
=
S E = N . 50 . 200 = N X 10.000/ mm
3
VOL =
1
5
1
20
1
10
X X
mm
3
= N
20 X
1
50
=
S E = N . 50 . 200 = N X 10.000/ mm
3
VOL (3 X 0.05 X 0.1 mm3) = 0.015 mm3
3 SQR = 3 X 0.015 = 0.045 mm3 N
S E = N
1
0.045
X 200 = N X
200
0.045
= N . 4.444
IMPROVED NEUBAUER
LEUKOCYTES COUNT :
4 CORNER SQUARES ( L ) = 1 X 1 X 0.1 mm
3

ERYTHROCYTES COUNT :
1
5
1
5
X X 0.1 mm
3
5 SQUARES ( E )
1
20
1
20
1
10
X X mm
3
80 SQUARES :
LEUKOCYTE COUNT
1. WIPE THE TIPS OF THE FINGER WITH COTTON AND
70% ETHANOL
2. PRICK THE FINGER WITH HEMOLANCET, WIPE AWAY
THE FIRST BLOOD
3. DRAW THE BLOOD UP USING THOMAS PIPET (LEUKOCYTE)
EXACTLY TO 0.5 MARK
4. DRAW THE TURK SOLUTION UP TO 11 MARK WITH
ROTATION SLOWLY
BLOOD DILUTID IS 1/20
5. MIX PIPET FOR APPROXIMATELY 3, BY PLACETHE PIPET
BETWEEN THUMB AND MIDDLE FINGER
MOVING YOUR HAND ONLY, DIRECTION IS RIGHT ANGEL TO
THE LONGITUDINAL AXIS OF THE PIPET
6. DISCARD THE FIRST 4 DROPS OF THE MIXTURE
7. FILL THE PREPARED COUNTING CHAMBER LEAVE + 3
ON THE BENCH
8. PLACE THE COUNTING CHAMBER ON THE STAGE OF
THE MICROSCOPE AND COUNT THE WHITE CELLS IN THE
APROPIATE SQUARE
DISCARD
TURK
SOLUTION
..
COUNT BY MICROSCOPE
CONDENSOR : DOWN WARD
OBJECTIVE : 10 X
INCLUDE IN THE COUNT THE CELL
SEEN ON THE LINES OF TWO SIDES
OF EACH SQUARES COUNTED AS
SHOWN IN FIGURE
CALCULATION :
BURKER COUNTING
CHAMBER
..
1
5
1
5
1
10
X X mm
3
25 SQR : N. CELLS
1
10
mm
3
N. CELLS S

1 mm
3

S

N X 10 X 20
S

N X 200 CELLS
NORMAL :
ADULT : 4.000 - 10.000 mm
3

INFANT NEW BORN : 10.000 - 12.000 mm
3
INFANT 1-6 MOUTH : 4.000 - 15.000 mm
3

CHILDREN 4 - 7 YEAR : 4.000 - 11.000 mm
3

8 - 12 YEAR : 4.000 - 10.000 mm
3
> N = LEUKOCYTOSIS BACTERIAL INFECTION
EXCEPT : S. TYPHOSA
< N = LEUKOPENIA INFECTION : S. TYPHOSA
MALARIA
IMPROVED NEUBAUER
COUNTING CHAMBER
LARGE CORNER SQUARES
1
10
mm
3
= N. CELLS 4 1 X 1 X
4
10
mm
3
= N. CELLS
1 mm
3
= N X X 20
4
10
= N X CELLS
200
4
- WHEN NORMOBLAST ARE PRESENT IN LARGE NUMBER CORRECTION
- COUNT THE NUMBER OF NORMOBLAST IN THE STAINED BLOOD FILM
FOR EVERY 100 WHITE CELLS
IF NORMOBLAST = 50
50
100 + 50
X 16 = 5.3 X 10
9
/L
LEUKOCYTE = 16 X 10
9
/L
LEUKOCYTE = 16 - 5.3 = 10.7 X 10
9
/L
FORMULATION N =
RED BLOOD CELLS COUNT (ERYTHROCYTE)
1. WIPE THE TIPS OF FINGER WITH COTTON AND 70% ETHANOL
2. PRICK THE FINGER WITH HEMOLANCET, WIPE AWAY THE 1
st
DROP
3. DRAW THE BLOOD UP USING A RED CELLS COUNT PIPETEXACTLY
TOTHE 0.5 MARK, WIPE OFF THE OUTSIDE OF THE PIPET CAREFULLY
4. DRAW THE HAYEM SOLUTION UP THE 101 MARK,
WITH ROTATION, BLOOD DILUTION 1/200
5. MIX FOR APPROXIMATELY 3 PLACE THE PIPET BETWEEN
THUMB AND MIDDLE FINGER, MOVING YOUR HAND ONLY WITH
DIRECTION VERTICAL TO THE LONGITUDINAL AXIS OF THE PIPET.
6. DISCARD THE FIRST FOUR DROPS OF THE MIXTURE
7. FILL THE PREPARED COUNTING CHAMBER, LEAVE 3 ON THE BENCH
8. PLACE THE CHAMBER ON THE STAGE OF THE MICROSPCOPE AND
COUNT RED CELLS IN THE APPROPIATE SQUARE
1
2
3
4
5
HAYEM
SOLUTION
6
7
COUNT BY MICROSCOPE
CONDENSOR : DOWN WARD
OBJECTIVE : 10 X /40X
CALCULATION
BURKER COUNTING
CHAMBER
..
IMPROVED NEUBAUER
COUNTING CHAMBER
COUNT IN THE RED CELLS IN
A. 80 SQUARES
1
20
1
20
1
10
X X mm
3

= N. CELLS
VOL = 80 X
1
20
1
20
1
10
X X = mm
3

1
50
1
50
mm
3
BLOOD
1
200
= N. CELLS
1 mm
3
= 50 X 200 X N. CELLS
(WHOLE BLOOD) = 10.000 X N. CELLS
B. 20 SQUARES
1
5
1
20
1
10
X X mm
3

=
1
50
mm
3
N. CELLS
1 mm
3
= 50 X 200 X N
= 10.000 X N
COUNT IN THE RED CELLS IN
5 SQUARES
1
5
1
5
1
10
X X
mm
3

= N.CELLS
VOL = 5 X
1
5
1
5
1
10
X X = mm
3

1
50
1 mm
3
= 50 X 200 X N. CELLS
(WHOLE BLOOD) = 10.000 X N. CELLS
NORMAL :
MEN 4.5 - 5.5 million/mm
3

WOMEN 4.0 - 6.0 million/mm
3

CHILDREN 4 YEARS 4.2 - 5.2 million/mm
3

INFANTS 1-6 MOUNTHS 3.8 - 5.2 million/mm
3

NEWBORN INFANT 5.0 - 6.0 million/mm
3
MICROCELL COUNTER
PRINCIPLE

- BLOOD CELLS SUSPENDED IN SALINE (0.95%) SOLUTION
STREAM THROUGH A CANAL CHINK ( 60 - 100 m m)

- EVERY CELL HAS ELECTRICAL RESISTANCE WHICH IS PROPOR-
TIONAL TO THE SIZE OF CELL

- THE ALTERATION OF THE ELECTRICAL RESISTANCE OCCUR,
FORM AN IMPULS FOR THE AUTOMATIC COUNTING APPARATUS
PREPARATION OF THIN BLOOD FILM
THIN BLOOD FILM (BLOOD SMEAR) IS USED TO DO :
1. DIFFERENTIAL COUNTING OF LEUKOCYTE
2.STUDY OF RED CELLS, WITH CELL AND PLATELET MORPHOLOGY
3.DETECTION OF BLOOD PARASITES
4. EXAMINE OF ANOTHER BLOOD DISORDER
MATERIALS :

1. SLIDES (2 PIECES), CLEAN, DRY AND FREE FROM FAT
2. 96% METHANOL
3. STANDARD GIEMSA SOLUTION
4. AQUADEST
METHOD
SMEAR OF BLOOD
TAIL
(THIN)
HEAD
THICK
1. PRICK THE FINGER TO GET BLOOD
2. COLLECT A DROP OF BLOOD AT ONE END OF FIRST SLIDE
3. PLACE THE EDGE OF 2
nd
SLIDE (SPREADER) JUST IN FRONT
OF THE DROP OF BLOOD
4. DRAW THE SPREADER BACKWARD UNTIL IT TOUCHES THE DROP
OF BLOOD
5. LET THE BLOOD RUN ALONG THE EDGE OF THE SPREADER
6. PUSH SPREADER TO THE OTHER END OF THE SLIDE, WITH
A SMOOTH MOVEMENT AND THE ANGLE OF ABOUT 30 - 45
0

7. WHEN THE BLOOD DROP IS ENOUGH, ALL THE BLOOD SHOULD
BE USED UP BEFORE YOU REACH THE END OF SLIDE
1
2
3
4
5
EVALUATION OF THE SMEAR
THE FILM IS SATISFACTORY
NOT GOOD
BLOOD DROP IS SMALL
BLOOD DROP IS TOO MUCH
SLIDE DO NOT FREE OF FAT
DRY IT


GEIMSA STAIN
FIXATION WITH 96% METHANOL 2 - 3
COVER THE SLIDE WITH DILUTED GIEMSA
STAIN (IN 1 : 10) 20
WASH THE STAIN OFF WITH BUFFER WATER
TIP THE WATER STAND THE SLIDE IN THE
DRAINING RACK TO DRY
DIFFERENTIAL CELLS (LEUKOCYTE) COUNT
TO DETERMINE THE RELATIVE NUMBER OF
EACH TYPE OF LEUKOCYTE,SPREADING AREA
OF LEUKOCYTE :
- PHERIPHERAL EDGE: THE LARGER LEUKOCYTE
= GRANULOCYTES & MONOCYTES
- CENTRAL OF THE SMEAR = LYMPHOCYTE METHOD :
TO PERFORM THE DIFFERENTIALCOUNTING
OF LEUKOCYTE MUST BE DONE AS :
MOVE THE SLIDE AS SHOWN IN FIGURE,AND
COUNT EACH LEUKOCYTE SEEN AND RECORD ON :
THE DIFFERENTIAL CELLS COUNTER OR ON A PIECE
OF PAPER UNTIL 100 CELLS
IF ANY NUCLEOTED RBC ARE SEEN ENUMERATED
THEM ON A SEPARATE COUNTER AND NOT TO BE
INCLUDED IN THE 100 CELLS DEFFERENTIAL COUNT
1 2 3 4 5 6 7 8 9 10 COUNT NORM
BASOPHILS - - - - - - - - - - - 0 - 1
EOSINOPHILS I - - - - - - - - II 3 1 - 3
STAB. II II - - - - - - - - 4 2 - 6
SEGMENT IIII I IIII IIII II IIII I IIII II IIII IIII I IIII IIII I IIII 58 50 - 70
LYMPHOCITE I II II IIII III IIII IIII IIII IIII III 33 20 - 40
MONOCYTE - I I - - - - - - - 2 2 - 8
COUNT 10 10 10 10 10 10 10 10 10 10 100
ABNORMAL FINDING :

NORMAL ABNORMAL

1. BASOPHILS 0 - 1 % BASOPHILIA -
2. EOSINOPHIL 1 - 3% EOSINOPHILIA ALLERGIC CONDITION
PARACITIC INVESTATION
SCARLET FEVER
LEUKEMIA
3. STAB CELL 2 - 6% SHIFT TO THE LEFT INFECTION
4. SEGMENT 50 - 70% NEUTROPHILIA APPENDICITIS ACUTE
BACTERIAL INFECTION
LEUKEMIA
5. LYMPHOCYTE 20 - 40% LYMPHOCYTOSIS VIRAL INFECTION
LEUKEMIA
6. MONOCYTE 2 - 8 % MONOCYTOSIS BRUCELLOSIS
TUBERCULOSIS
LEUKEMIA
1. BASOPHIL CELL
- SIZE : 11 - 13 m m
- SHAPE ROUND (DAMAGED)
- NUCLEUS : DIFFICULT TO SEE
BECAUSE COVERED BY GRANULES
- CYTOPLASMS : ABUNDANT
- GRANULES VERY LARGE ROUND
DEEP PURPLE, NUMEROUS
2. EOSINOPHIL CELL
- SIZE : 12 - 15 m m
- SHAPE ROUND
- NUCLEUS : USUALLY 2 LOBES
- CYTOPLASMS : LARGE
- GRANULES : LARGE, ROUND, ORANGE
RED, NUMEROUS, CLOSELY
PACK
3. NEUTROPHIL CELL
A. STAB CELL
= IMMATURE CELL (BAND FORM)
- SIZE 12 - 15 m m
- SHAPE = ROUND, WELL DEFINED
- NUCLEUS = BAND SHAPE
- CYTOPLASMS = ABUNDANT
PINKISH
- GRANULES = MAUVE, VERY SMALL
NUMEROUS BUT
SEPARATE

3. B. SEGMENT NEUTROPHIL CELL
SIMILAR TO PREVIOUS LEUKOCYTE
EXCEPT THAT
THE NUCLEUS : SEVERAL (2-5), LOBES,
LINKED BY STRANDS OF
CHROMATINE
THE CHROMATINE APPEAR AS UNIFORM
DEEP PURPLE MASS

4. LYMPHOCYTE CELL
A. SMALL LYMPHOCYTE CELL
- SIZE : 7 - 10 m m
- SHAPE : ROUND
- NUCLEUS : LARGE, OCCUPYING MOST
OF THE CELL, CHORMA-
TINE DENSE, DARK PURPLE

B. LARGE LYMPHOCYTE CELL

- SIZE : 10 - 15 m m
- SHAPE : ROUND AND IRREGULAR
- NUCLEUS : OVAL OR ROUND
MAY LIE ON ONE SIDE
OF THE CELL
- CYTOPLASMS : ABUNDANT
PALE BLUE
5. MONOCYTE CELL
- SIZE : 15 - 25 m m
- SHAPE : IRREGULAR
- NUCLEUS : VARIBLE, OFTEN
KIDNEY SHAPED/
LOBED (2-3 LOBI)
CHROMATINE ARRANGED
IN STRANDS, PALE MAUVE
- CYTOPLASMS : ABUNDANT
- GRANULES : FINE, DUST LIKE
USUALLY REDDISH
- VACUOLES : PRESENT IN THE CYTOPLASM
ERITHROCYTE SEDIMENTATION RATE
(ESR)
PRINCIPLE : ANTICOAGULATED WHOLE BLOOD IS ALLOWED TO
STAND FOR A PERIOD OF TIME,
THE ERITHROCYTE WILL SETTLE OUT FROMTHE PLASMA
THE RATE AT WHICH THE ERITHROCYTE FALL IS
KNOWN AS ESR
BLOOD
+
ANTICOAGULANT
ALLOWED TO
STAND FOR
PLASMA
ERITHROCYTE
METHODS : 1. WINTROBE METHOD
2. WESTERGREN METHOD
1. WINTROBE METHOD
BLOOD : Na.CIT.
( 4 : 1 )
MATERIALS :
- SYRINGE AND NEEDLE
- TUBE/ BOTTLE AND ANTICOAGULANT
SODIUM CITRATE
- PASTEUR PIPET
- THE WINTROBE TUBE
PASTEUR PIPET
STAND FOR
1 HOUR
METHOD :
1. PLACE IN A TUBE OR BOTTLE 0.4 ml OF TRISODIUM CITRATE
SOLUTION
2. COLLECT VENOUS BLOOD :
- APPLY TOURNIQUET AS LOOSE AS POSSIBLE
- PUNCTURE THE VEIN AT ONCE RELEASE THE TOURNIQUET
COLLECT INTO THE SYRINGE 2 ml BLOOD AND PUT INTO
TUBE OR BOTTLE WITH ANTICOAGULANT
1.6 mL BLOOD, SHAKE GENTLY
3. FILL THE BLOOD INTO THE WINTROBE TUBE USING A
PASTEUR PIPET UP TO 0 MARK
4. PLACE THE TUBE UP RIGHT AND ALLOWED TO STAND FOR
1 HOUR
5. READ THE HEIGHT OF THE COLUMN OF PLASMA IN mm
GRADUATION STARTING FROM THE 0 MARK
NORMAL :
ESR : MEN : 0 - 10 mm / HOURS
WOMEN : 0 - 20 mm / HOURS
2. WESTERGREN METHOD :
MATERIALS : 2.0 mL SYRINGE + NEEDLE
3.8 % SODIUM CITRATE
WESTERGEN ESR TUBES
WESTERGEN STAND
ANTICOAGULANT (3.8%)
METHOD :
1. PLACE IN A BOTTLE 0.4 mL OF SODIUM CITRATE SOLUTION
2. COLLECT VENOUS BLOOD
3. ADD 1.6 mL BLOOD TO THE BOTTLE CONTAINING ANTICOAGULANT
4. SHAKE GENTLY
5. DRAW THE CITRATE BLOOD INTO THE WESTERGREN TUBE UP
TO THE 0 MARK
6. PLACE THE TUBE IN THE WESTERGREN STAND UP RIGHT
7. WAIT FOR ONE HOUR
NORMAL : MEN : 0 - 15 mm / HOURS
WOMEN : 0 - 20 mm / HOURS
CHILDREN : 0 - 10 mm / HOURS
PATHOLOGIC : - ACTIVE TBC
- RHEUMATIC FEVER
- MALIGNANT DESEASE
NOTES : THE HEIGHT OF THE COLUMN OF PLASMA IN mm
GRADUATION STARTING FROM THE 0 MARK AT
THE TOP OF THE TUBE

1.6 mL BLOOD
0.4 ml 3.8% SOD. CITRATE
SHAKE
6. THROMBOCYT COUNT
6A. DIRECT METHOD
MATERIALS :
- PIPET THOMA FOR ERYTHROCYTE
- 1 % AMM. OXALAT SOLUTION
- HAEMOCYTOMETER (IMPROVED NAUBAUER)
METHOD :
- TAKE THE BLOOD WITH RED PIPET TO THE 1.0 AND
AMM. OXALAT 101, DILUTION 1/100, MIX CAREFULLY
- DISCARD FIRST FOUR DROP, PLACE A DROP OF MIXTURE
ON THE HEMOCYTOMETER AND PLACE IT THE TO STAND FOR
15 MINUTE INSIDE TO PETRI PLATE WITH CONTAIN MOIST
FILTER PAPER
- COUNT THE PLATELET WITH PHASE MICROSCOPE
IN 80 SQUARES :
1
20
X
1
20
1
10
X mm
3

WHEN
1
50
mm
3
= N. CELLS
THE COUNT OF PLATELETS IN 1 mm
3
= N X 50 X 100
= N X 5000 CELLS
6B. INDIRECT METHOD (FONIO)
I.
GIEMSA STAIN
MATERIALS :
1. 14 % SOLUT. MgSO
4

2. BLOOD LANCET
3. SLIDES
4. GIEMSA SOLUTION
METHOD :
1. DROP SOLUTION 14 % MgSO
4
ON THE FINGER TIP
2. PRICK AND MIX WITH BLOOD
3. MAKE BLOOD SMEAR AND STAIN BY GIEMSA STAIN
COUNT : ERITHROCYTE 1000 CELLS
THROMBOCYT : N. CELLS
II. MAKE THE ERITHROCYTE COUNT
RESULT = Y. CELLS / mm
3

THE COUNT OF THROMBOCYT =
N x Y
1000
/mm
3

NORMAL RANGE = 150.000 - 400.000 / mm
3

INCREASE = THROMBOCYTOSIS : - POLYCYTHEMIAVERA
- IDIOPATHIC THROMBOCYTHEMIA
- CHRONIC MYELOGENIUS LEUKEMIA
- FOLLOWING SPLENECTOMY

DECREASE = THROMBOCYTOPENIA : -THROMBOCYTOPENIA PURPURA
- APLASTIC ANEMIA
- ACUTE LEUKEMIA
- GAUCHERS DISEASE
- PERNICIUS ANEMIA
- FOLLOWING CHEMO/RADIATION
THERAPY
7. RETICULOCYT COUNT
RETICULOCYT IS A YOUNG ERITHROCYTE WHICH CONTAIN :
- A BASOPHYLIC SUBSTANCES NAMED SUBSTANSIA GRANULO-
FILAMENTOSA
MATERIALS : - 2 PIECES OBJECT GLASS
- SMALLTUBE
- DROP PIPET
- 1 % SOLUTION BRILLIANT CRESYL BLUE
(OR NEW METHYLEN BLUE SOLUTION)
METHOD :
PRICK FINGER
TO GET BLOOD
OR EDTA VENOUS
BLOOD
MIX
BLOOD 3 DROPS
BCB SOL. 3 DROPS
COTTON
STAND FOR 15
AND 37
0
C
MAKE PREPARATION
MOIST
DRY
EXAMINE WITH MICROSCOPE
DIAFRAGMA IN EYE PIECE
FROM PAPER WITH APERTURE
COUNT :
ERYTHROCYTE = 1000 CELLS
RET : N. CELLS
IT MEANS THE COUNT OF
THROMBOCYT IS (N %O )
ERY : PALE BLUE
RET : SIZE > E
GRANULES / RET
FINE/VIOLET
NORMAL : ADULT : 0.2 - 2.0 %
(2 - 20 %O)
BABY : 0.2 - 6.0 %
(2 - 60 %O)
8. PCV (PACKED CELL VOLUME) / HEMATOCRIT
TO ESTIMATE VOLUME OF ERYTHROCYTE AGAINTS BLOOD
METHODS : 1. MACROHEMATOCRIT
2. MICROHEMATOCRIT
1. MACROHEMATOCRIT
MATERIAL : 1. SYRINGE + NEEDLE
2. WINTROBE TUBE
3. PASTEUR PIPET
4. SMALL BOTTLE + ANTICOAGULANT,
EDTA, HEPARIN
METHOD :
CENTRIFUGE
3000 RPM
15 - 30
PLASMA
LIPID
BUFFYCOAT
1 mm 10.000 PLAT/mL
ERYTHROCYTE
45
PCV = 45 %
HEMATOCRIT IN % =
100 X HEIGHT OF RED CELL (mm)
HEIGHT OF THE WHOLE BLOOD (mm)
HEMATOCRIT : HIGH ALTITUDE >
AT BIRTH : 50 - 62%
1 YEAR : 31 - 39%
ADULT WOMEN : 36 - 46%
ADULT MEN : 42 - 52%
BLOOD EDTA
2. MICROHEMATOCRIT
SPECIMEN :
CAPILLARY BLOOD
WHOLE BLOOD WITH EDTA
MATERIAL :
- CAPILLARY HEMATOCRIT TUBES
- HEMOLANCET + COTTON 70% ETHANOL
- CREATOSALE (SOFT WAX)
- CENTRIFUS MiH
- READING DEVICE
CREATOSEAL
2 mm
2/3 - 3/4

1. PRICK THE TIP OF FINGER
WITH HEMOLANCET
2. FLOW THE BLOOD INTO
THE CAPILLARY TUBE
FILL 3/4 OF TUBE
3. PLUG THE END OF TUBE
WITH CREATOSEAL
4. CENTRIFUGE :
- 10.000 - 20.000 RPM
- 5
1
2
3
READING
DEVICE
TOP OF PLASMA
TOP OF RBC
THE BOTTOM OF RBC
READ : USING READING DEVICE
NORMAL : MEN : 40 - 54%
WOMEN : 37 - 47%
METHOD :
P.C.V IS NEEDED TO ESTIMATE ABSOLUTE VALUE OF BLOOD :
M.C.V. (MEAN CORPUSCULAR VOLUME)
M.C.H.C. ( MEAN CORPUSCULAR HB CONCENTRATION)
PCV
S. ERI
X 10 m
3
(76 - 96 m
3
) =
fl = FENTOLITER
Hb
PCV
X 100% (32 - 36%

) =
PERCENT
M.C.H. ( MEAN CORPUSCULAR HB)
Hb
S. ERI
X 10 m m g

(27 - 32 m m g

) =
pg = PIKOGRAM
THIN BLOOD FILM ARE USED :

1. DIFFERENTIAL COUNTING
2. STUDIES OF MORPHOLOGIC BLOOD CELLS
3. IDENTIFYING PARASITES
- NORMAL : SIZE UNIFORM
SHAPE ROUND BICONCAVE
FROM ABOVE FROM SIDE
F 6 - 9 m m (7 m m)
EVALUATION OF MORPHOLOGIC BLOOD CELLS
1. ABNORMALITIES OF SIZE :
ANISOCYTOSIS
A. MICROCYTIC
4 - 6 m m
B. MACROCYTIC
10 - 12 m m
C. SHCIZOCYT
(FRAGMENT)
D. MEGALOCYTIC
( 12- 25 m m)
2. ABNORMALITIES OF SHAPE
(POIKILOCYTOSIS)
A. TARGET CELLS
B. CRENATED CELLS
C. SPHEROCYT
D. ACANTHOCYT
E. BURR CELL
F. CRESCENT BODY
G. OVALOCYT /
ELLIPHTOCYT
H. SICKLE CELL
I. TEAR DROP APP.
J. LEPTOCYT
K. BIZZARE
ERYTHROCYTE
NORMOCHROM NORMOCYTIC
3. ABNORMALITY OF COLOUR :
A. HYPOCHROM
B. HYPERCHROM
C. POLYCHROMATION
D. STOMATOCYTE
4. INCLUSION BODY
A. BASOPHYLIC STIPLING
B. MALARIAL STIPLING
C. SIDEROSIT
D. CABOTS RING
E. HOWEL-JOLLY BODY
ABNORMAL MORPHOLOGIC OF LEUKOCYTE
1. HYPERSEGMENTATION
( 5 OR MORE SEGMENT)
2. DEGENERATED NEUTROPHILS
WITH PIKNOTIC NUCLEUS
3. KARYOREXIS
4. TOXIC GRANULES
(INFECTION POISONING BURN)
5. VACUOLIZATION
6. DOHLES (INCL.) BODIES
INFECTION POSITION BURN
7. AUER RODS
8. BARR BODY
9. PELGER-HUET ANOMALY
( < 2 LOBES)




CRENATED ROULEAUX SMUDGE
CELL FORMATION CELL
ARTEFACT :
BLOOD PARASITES
MALARIAL PARASITE
THIN BLOOD FILM
THICK BLOOD FILM
DRY
FIXATION WITH
96% ETHANOL
DRY
DROP WITH WATER : HAEMOLYSIS
STAIN BY GIEMZA FOR 30
RINSE WITH AQUADEST / BUFFER
DRY
MICROSCOPE EXAMINATION
LOOK FOR : PL ; VIVAX, MALARIAE, FALCIPARUM & OVALE
VIVAX
MALARIAE
FALCIPARUM
OVALE
TR TR TR
SCHIZONT
GAMET
CAPILLARY FRAGILITY TEST
(RUMPLE LEEDE TEST)
WITH THE SPHYGMOMANOMETER,
MEASURE THE SYSTOLIC AND DIASTOLIC
BLOOD PRESURE, THEN PRESS THE HAND
WITH PRESSURE OF 1/2 (SYS + DIAST) FOR
10 . ON THE FORHAND ABOUT4 cm BELOW
THE ELBOW, MAKE A CIRCLE WITH DIA-
METER 5 cm.
EXAMINE AND COUNT THE PETERCHIAE
ESPECIALLY IN THE CIRCLE
RL + = IF THE CIRCLE CONTAIN MORE
THAN 10 PETERCHIAE
1/2 (S + D)
4 cm
BLEEDING TIME TEST
A. DUKE METHOD
MATERIAL :
1. HEMOLANCET AND
COTTON + ETHANOL 70%
2. STOP WATCH
3. FILTER PAPER

METHOD :
1. PUNCTURE THE EAR LOBE DEEPLY
2. START STOP WATCH
3. EVERY 30 SECOND, COLLECT THE DROP
OF BLOOD ALONG THE STRIP OF
BLOTTING PAPER
4. WHEN NO MORE BLOOD APPEAR,
STOP THE STOP WATCH
5. RESULT CAN MADE :
A. THE TIME OF WATCH
B. COUNT THE NUMBER OFF DROP &
MULTIPLY BY 30
NORMAL : 1 - 3 MINUTE
LOCALIZATION :
- FOREHAND
- PLACE BLOOD PRESSURE CUFF
PRESS UP TO 40 mm Hg
- MAKE 3 mm DEEP SKIN PUNCTURE
START :
- BLOT THE BLOOD FROM PUNC-
TURE SITE ON A PIECE OF CIR-
CULAR FILTER PAPER EVERY 30
SECOND WHEN BLEEDING CASES,
STOP THE WATCH AND RELEASE
THE BLOOD PRESSURE CUFF
- RECORD THE BLEEDING TIME
NORMAL : 1 - 7 MINUTE
x
40 mmHg
EVERY
30 SECOND
EVERY
30 SECOND
B. IVY METHOD
1
2
COAGULATION TIME OF WHOLE BLOOD
(LEE & WHITE)
3 ml
1 ml
1 ml
f : 7 - 8 mm
- AFTER 3, REMOVE THE 1
st
TUBE
AND TILT FOR 45
0

- REPEAT EVERY 30 SECOND
UNTIL THE TUBE CAN BE
COMPLETELY INVERTED
- C.T. DETERMINED BY CLOTTING
TIME OF TUBE NO. 2
GLASS TEST
TUBE
1 2
WATER BATCH 37
0
C
NORMAL : 5 - 11 MINUTE
CLOT RETRACTION
> 5 ml
WATER BATCH 37
0

1 HOURS
CLOT
TEST :
- THROMBOCYTE FUNCTION NUMBER
- FIBRINOGEN CONCENTRATION
- PACKED CELL VOLUME
C.R. = % ACE SERUM
FROM BLOOD VOLUME
IF THIS SERUM = 3 ml
BLOOD = 5 ml
C.R. = 3/5 X 100 = 60%
NORMAL : 40% - 60%
N = CR OCCURED AT 2 - 24 HOURS
POOR = CR OCCURED AFTER 4 HOURS WITHIN 24 HOURS
NIL = NO RETRACTION OCCURS AFTER 24 HOURS
START THE STOPWATCH
AS SOON AS THE BLOOD
ENTERS
THE SYRINGE
BLOOD GROUPING
SLIDE TEST :
ANTI A ANTI B ANTI A+B
MIX & SHAKE
AGGLUTINATION :
+ - + A
- + + B
+ + + AB
- - - O
TILE TEST :
+
ERY A
ERY B
ERY X
SERUM X ANTI A
- +
+ - +
- + +
ANTI B
BLOOD GROUP
+
ERY A
ERY B
ERY X
SERUM X ANTI A
+ +
+ - +
- + +
ANTI B
+ + +
+ + +
+ + +
+ + +
+ + +
+ + +
L L
L
+ + +
+ + +
+ + +
1. FAECES COLLECTION :

A. COLLECT FAECES IN A DISPOSIBLE CONTAINER
B. DO NOT MIX WITH URINE
C. EXAMINE A FAECES WITHIN 30 - 40
IF NOT, PLACE IT IN REFRIGATOR
D. PATIENT RECOMENDED NOT TO EAT SUBSTANCE
BARIUM, BISMUTH IN 5 DAYS
E. WHEN THERE IS OBSTIPATION CATHARTIC NaCl/Na
2
SO
4



NOTES :

USE FAECES RANDOM, RARELY 24 HOURS
IMPORTANT EXAMINATION OF FAECES :
- EGGS OF WORM
- PARASITES
- OCCULT BLOOD
SELECT THE PORTION OF FAECES WHICH GIVE LARGE
POSSIBILITY TO FIND ABNORMALITIES

EXAMINATION
OF FAECES
MACROSCOPIC
1. FORM
2. COLOUR
3. SMELL
4. CONSISTENCY
5. MUCUS
6. BLOOD
7. PUS
8. REST. OF FOOD
9. PARASITE
MICROSCOPIC
1. REST OF FOOD
2. ERITHROCYTE/
LEUKOCYTE
3. AMOEBAE
4. THE EGGS OF WORMS
- CARBOHYDRATE
- PROTEIN
- FAT
- VEGETABLES
CHEMIST
1. BLOOD
2. STERCOBILIN
EXAMINATION OF FAECES
A. MACROSCOPIC EXAMINATION OF THE FAECES
(FRESH FAECES)
1. FORM : A NARROW / RIBBON LIKE FAECES
2. THE COLOUR OF FAECES :
A. NORMAL : BROWNISH / PALE BROWN
B. ABNORMAL :
1) BLACK : MELAENA = BLEEDING FROM THE UPPER GUT
2) CLAY COLOUR : ACHOLIS = DIMINUTION / ABSCENT
OF STERCOBILIN
3) RED = BLOOD, ORIGINATING FROM THE LOWER GUT
= FOOD / BEETS
4) THE OTHER : VEGETABLES DIET
- GREEN
- YELLOW

3. SMELL OF FAECES :

- NORMAL : SMELL WITH INDOL,SKATOL, BUTIRIC ACID
- PUTRESCENT : PUTREFACTION OF PROTEIN BY BACTERIA
- ACID : CARBOHYDRATE FERMENTATION = DIARE
- ROTTEN : FAT FATTY ACID
- RANCID : BACILLARY DYSENTRI
4. CONSISTENCY OF FAECES :

A. NORMAL : SOFT
B. ABNORMAL :
- HARD : CONSTIPATION
- LIQUID : PAPPY FORM (DIARRHE)
WATERY (CHOLERA)
- + (CO
2
) : FERMENTATION OF CARBOHYDRATE
INFLAMMATION / STIMULATION ON INTESTINE WALLS
TRANSLUCENT GELATINOUS MUCUS :
- SPASTIC CONSTIPATION
- MUCOUS COLITIS
BLOODY MUCUS : INFLAMMATION
MUCUS + BLOOD + PUS : ULCERATIVE COLITIS, BACILLIARY,
DYSENTRY
OUT OF THE FAECES : FROM LARGE INTESTINE
MIX MUCUS : FROM SMALL INTESTINE
5. MUCUS
6. BLOOD :
- FRESH BLOOD, FAECES LIQUID DYSENTERIAE
FRESH BLOOD, FAECES NORMAL ANAL FISSURES
HAEMORRHOID
- BLACK : MELAENA
7. PUS :
- LOCALIZED ABSCESSES BROKEN PUS IN FAECES
8. REST OF FOOD :
- VEGETABLES
- GRAINS LEGUMINOCEAE, ETC.
9. PARASITE :
- ASCARIS LUMBRICOIDES (ROUND WORM)
- ENTEROBIUS VERMICULARIS
B. MICROSCOPIC EXAMINATION OF THE FAECES
1. REST OF FOOD :
A. CARBOHYDRATE : LUGOL SIDE
B. PROTEIN : ACETIC ACID SLIDE
C. FAT : SUDAN III SLIDE
D. VEGETABLES : ACETIC ACID SLIDE
2. AMOEBA / OTHER PROTOZOA : EOSIN SLIDE
3. ERYTHROCYTE / LEUKOCYTE : EOSIN SLIDE
4. THE EGGS OF WORM : - DRY SLIDE METHOD
- CONCENTRATION TEST
PREPARATION FOR THE TEST
MAKE :
1. SUSPENSION
OF FAECES
2. REAGENT FOR TEST
(EOSIN, ETC)
MIX
PUT THE COVER GLASS ON IT
EXAMINE BY MICROSCOPE
MICROSCOPIC EXAMINATION OF FAECES
1. REST OF FOOD
A. CARBOHYDRATE : LUGOL
AMYLUM / CH + IODIUM BLUE
CARBOHYDRATE
DEXTRIN
REAGENS
B. 30 % ACETIC ACID RED MUSCLE
VEGETABLE
CONECTIVE
TISSUE
C. FAT SUDAN III
RED BALL STRUCTURE
2. AMOEBA + EOSIN AS NEGATIVE STAINING
V
K
L
E
(BACKGROUND : RED COLOUR)
AMOEBA
ERY / LEUKO
COLOURLESS
3. ERYTHROCYTE / LEUKOCYTE : EOSIN (1 - 2%)
MORPHOLOGY APPEAREANCE
SAME LIKE IN THE SEDIMENT OF URINE
4. THE EGGS OF WORM
A. DRY PREPARATION

MATERIALS :
- OBJECT GLASS
- PARAFIN LIQUIDIUM
- WOOD STICK
STREAK THE FAECES
ALLOW TO DRY ALL
OF THE FAECES
COVER WITH
PARAFIN LIQUIDIUM
THROUGH OUT THE FAECES
MICROSCOPIC
EXAMINATION
ADVANTAGE OF USING PARAFIN LIQUIDUM :
1. THE SLIDE IS CLEAN AND NOT DIRTY
2. THE SLIDE IS GOOD SMELLING
3. THE PANORAMA IN THE MICROSCOPE IS MORE CLEAR
MAKE ATTENTION TO THE EGGS OF WORM OF :
ASCARIS
LUMBROCOIDES TRICHOCEPHALUS
ANKYLOSTOMA
DUO DENALE
B. CONCENTRATION TEST
ESPECIALLY FOR DETECTION THE EGG OF ANKYLOSTOMA
PRINCIPLE :

SPECIFIC GRAVITY OF THE EGG IS LESS THAN THE SPECIFIC
GRAVITY OF NaCl SOLUTION SO THAT THE EGGS ARE FLOATING
ON THE SOLUTION AND ATTACHED TO THE COVER GLASS
1. MAKE SUSPENSION OF 5 gr FAECES IN CONCENTRATE NaCl
SOLUTION IN GLASS TUBE
2. ADD SOLUTION UNTILL THE GLASS TUBE IS FULLFILLED
WITH SUSPENSION
3. PUT A COVER GLASS ON THE TOP OF GLASS TUBE
4. ALLOW TO STAND FOR 1/2 - 1 HOUR
5. LAY A COVER GLASS ON AN OBJECT GLASS IN FACE DOWN
POSITION
6. EXAMINE BY MICROSCOPE
METHOD :
MICROSCOPE
C. CHEMICAL EXAMINATION
1. OCCULT BLOOD EXAMINATION
SMALLER INCREASES IN BLOOD CONTENT MAY NOT ALTER
APPEAREANCE OF THE STOOL
REAGENTS :

1. BENZIDINE
2. ACETIC ACID GLASIALE
3. H
2
O
2
3%
FAECES SUSPENSION
METHOD :
ACETIC ACID 3 ml
1 gr
BENZIDIN
FILTER
FILTRAT
H
2
O
2
3%
3 ml
: GREEN TO BLUE
+
READ WITHIN 5
BENZIDIN TEST :
- VERY SENSITIVE = 10 - 1000 TIMES MORE SENSITIVE THAN
GUAIAC TEST
- THE PATIENT SHOULD HAVE DONE THE BENZIDINE DIET
- OR A MEAT FREE DIET FOR 3 - 5 DAYS
1.A. BENZIDINE TEST
1.B. GUAIAC TEST
GUAIAC IS 20 TIMES LESS SENSITIVE THAN BENZIDINE
GIVE + RESULT WHEN 24 HOURS FAECES CONTAIN A HALF
ml OF BLOOD
GUAIAC HARS IS DIFFICULT TO GET, NEVER BE DONE
REAGENTS :
1. 1 : 60 SOLUTION OF GUM GUAIAC IN 96% ETHYL ALCOHOL
2. GLACIALE ACETIC ACID
3. 3% H
2
O
2
PROCEDURES :
MIX
WELL
MIX
WELL
MIX
WELL
OBSERVE FOR 2 MINUTE
RECORD THE MAXIMAL BLUE COLOUR
2 ml WATER
0.5 gr FAECES
0.5ml
Gl.A A
2 ml GUAIAC
SOLUTION
MIX
STARTING
TIMER
2 ml H
2
O
2
3%
TRACE 1 + 2 ++ 3 +++ 4 ++++
2. STERCOBILIN
5 ml HCl
CONCENTRATE
FAECES 1 - 3 gr
STAND FOR
SEVERAL HOURS
(ONE NIGHT)
BOIL
READ SOON
UROBILIN
PALE RED
BILIRUBIN
BLUE COLOUR
SPUTUM
BE CAREFUL, BECAUSE THIS SPECIMENT (SPUTUM) IS INFECTIOUS
THE METHOD OF HANDLING AND DISCHARGE MUST BE STERILIZE
OR BURNED
= COUGH SECRETE FROM BRONCHEAL TREE SINCE THE SPUTUM
IN EVITABLY ARE CONTAMINATED WITH SALIVA, ETC
THE COLLECTION OF SPUTUM MUST BE CARRIED OUT WITH
SUPERVISED BY PROFESIONAL PERSON
SPUTUM CAN BE COLLECTED AT THE TIME THAT NEEDED OR
THE FIRST MORNING SPUTUM IS BETTER ESPECIALLY FOR
MICROBIOLOGICAL EXAMINATION.
OCCASIONALLY : - 12 HOURS SPUTUM
- 24 HOURS SPUTUM
COLLECT IN A STERIL, DISPOSIBLE, INPERMIABLE WIDE MOUTH
CONTAINER :
- PETRI PLATES
- BOTTLE
- SPUTUM CARTOON
EQUIPMENT :

- WORKING TABLES
- MICROSCOPE
- AND OTHERS
SHOULD HAVE BEEN CLEANED
WITH DISINFECTANT LIKE
10% LYSOL SOLUTION
MACROSCOPIC EXAMINATION
1. VOLUME : VARIES WITH THE NATURE AND EXTEND OF THE
PATHOLOGIC PROCESSES
- NORMAL : - / + SMALL VOLUME
- LARGE VOLUME : = EDEMA PULMONARY
(100-500 ml/24 hours) = ABSCESS PULMONARY
= BRONCHIECTASIS
= CAVITARY PULMONARY TUBERCULOSIS
= LIVER ABSCESS PENETRATE TO THE LUNG
2. SMELL : FRESH SPUTUM ODOUR
- ROTTEN SMELL = GANGREN PULMONAL & ABSCESSES
= NECROTIC TUMOR
= EMPYEMA : BRONCHI
- THE ODOUR OF FAECES : SUBPHRENICAL ABSCESS PENETRATE
TO THE LUNG

3. COLOUR :
- GRAY : PUS & EPHITELS
- RED : FRESH BLOOD
- BROWNISH RED : OLD - BLOOD = PNEUMONIC LOBE
OR GANGREN
- BLACK : DUST
4. CONSISTENTION : VARIES WITH KIND AND STAGE OF DISEASE

- SEREOUS : PULMONARY EDEMA
- MUCOUS : BRONCHITIS, ASTHMA, PN.LOBARIS
- PURULENT : PULMONARY ABSCESS, BR. ECTASIS
- MIX : @ SERO - PURULENT
@ MUCO - PURULENT
@ SERO - HEMORAGIC
24 HOURS SPUTUM WHICH PERMITTED TO SETTLE
MOST VISCID
LESS VISCID
VISCID
PORTION
SECOND
FIRST
THIRD
FOAMIC LAYER
CLOUDLY FLUID LAYER
CONSIST OF PUS, TISSUE,
BACTERIA, ETC.
BRONCHIETASI
GANGRENE
PULMONAL ABSCESS
VISCID SPUTUM : KLEBSIELLA PULMONARY
RUSTY SPUTUM : PNEUMOCOCAL PNEUMONIA
HEMAPTYSIS : CARDIAC FAILURE
PULMONARY INFARCTION
PROGRESSIVE INFECTION
NEOPLASM
5. SPECIFIC ELEMENT
EXAMINE WITH HAND LOUPE
A. CHEESE GRANULE = NECROTIC TISSUE :
- KOCH PULMONUM
- GANGRENE
- PULMONAL ABSCESS
- ACTINOMYCOSIS
B. SPIRALE OF CURCHMANN : ASTHMA BRONCHIALE
C. BRONCHIAL CAST : BR. TIS FIBRINOSA
PNEUMONIA
MICROSCOPIC EXAMINATION
NATIVE SLIDES
STAINING OF :
- GIEMSA : BLOOD, CYTOLOGY
- GRAM : BACTERIOLOGY
- ZIEHL NEELSEN, ACID FAST BACILLI
TRANSUDATE & EXUDATE
BODY CAVITY
CEREOUS
FLUID
PLASMA
ULTRAFILTRATION
VOLUME IS INFLUENCED BY :
- OSMOTIC AND HYDROSTATIC PRESSURE OF BLOOD
- PLASMIC CHEMICAL ELEMENT CONCENTRATION
- CAPILLARY PERMIABILITY
IN THE PATHOLOGIC CONDITION : THE VOLUME CAN BE INCREASED
THE DIFFERENT BETWEEN :
TRANSUDATE EKSUDATE
INFLAMATION NO YES
STERILITY STERILL NO STERIL, BACTERIA +
CLEARNESS CLEAR TURBID, PURULENT BLOOD +
SPEC. GRAVITY < 1.018 > 1.018
PROTEIN < 2.5 g % > 2.5 g %
CELL COUNT SMALL AMOUNT ( < 500) LARGE AMOUNT (> 500)
COAGULATE - (MINUS) + (PLUS)
GLUCOSE JUST SAME WITH BLOOD LESS THAN BLOOD GLUCOSE
RIVALTA TEST - (MINUS) + (PLUS)

THE KIND OF TRANSUDATE & EXUDATE
- THE FLUID OF : - PERITONEAL CAVITY
- PLEURAL CAVITY
- PERICARDIAL CAVITY
- SYNOVIAL CAVITY
- HYDROCELE
- ETC.
THE METHOD OF COLLECT SAMPLE :

PUNCTION :
UNKNOWN
TRANSUDATE (INFLAMATION NEG. -)
OR
EXUDATE (INFLAMATION POS. +)
ASEPTIC PUNCTION + ADD BY ANTICOAGULANT ( 20% SODIUM CITRATE)
ROUTINE & CHEMIST EXAMINATION BACTERIOLOGIC EXAMINATION
IN LABORATORY
MACROSCOPIC EXAMINATION
1. COLOUR : - YELLOWISH
- GREENISH YELLOW
- CREAM COLOURED
- RED, ETC.
2. SMELL
3. CLEARNESS : - SEROUS
- SERO-FIBRINOSA
- SERO-SANGUINOSA
- PURULENT
4. COAGULATE : SMOOTH, SHREADS, CLOT, ETC
5. SPECIVIC GRAVITY : ESTIMATE WITH URINOMETER
MICROSCOPIC EXAMINATION
1. BLOOD CELLS COUNTING
FLUID + ANTICOAGULANT
MAKE DILUTION WITH SALINE (0.9 % NaCl)
2. DIFFERENT COUNT : LIMPHOCYTE / PMN
SEDIMENT GIEMZA SLIDE COUNT (100 - 300 CELLS)
3. PAPANICULAOU STAIN ABNORMALLY CELLS
4. GRAM / ZIEHL NEELSEN STAIN BACTERIOLOGIC
CHEMICAL EXAMINATION OF TRANSUDATE AND EXUDAT
= LIMITED FOR PROTEIN AND GLUCOSE
A. QUALITATIVE TEST FOR PROTEIN CONCENTRATION

METHOD :
1 cm
FLUID
(EXUDATE / TRANSUDATE)
AQUADEST
+
1 GTT GLACIALE ACETIC ACID
RECORD THE REACTION
TURBIDITY OF SOLUTION
RESULT :

= CLEAR (NOT TURBID)


= RATHER TURBID (FINE)
(WEAK)

= REAL TURBID, PRECIPITATE
(STRONG) DENSE FOG,
-
+
+
- THE TEST MUST BE REPEATED
- BASIC : SERO.MUCIN +
B. QUANTITATIVE TEST FOR PROTEIN CONCENTRATION
1. DEFINITE THE SPECIAL GRAVITY OF LIQUID
WHEN S.G. < 1.010 DILUTE 5 - 1 0 x
S.G. > 1.010 DILUTE 20 x
2. MAKE ESBACH TEST :
FORMULATION : (S.G. - 1.007) X 343 = gr / 100 ml PROTEIN
S.G. 1.010 1 gr / 100 ml PROTEIN
S.G. 1.015 2.5 gr / 100 ml PROTEIN
S.G. 1.020 4.5 gr / 100 ml PROTEIN
S.G. 1.025 6 gr / 100 ml PROTEIN
RIVALTA TEST
CEREBRO SPINAL FLUID (CSF)
= PLASMA ULTRAFILTRATION
FORM BY ACTIVE SECRETION IN THE FLEXUS CHOLORIDEUS
AND DIFFUSION FROM PLASMA
CSF USUALLY IS OBTAINED BY PUNCTURE OF CAVUM
SUBARACH NOIDALE AT :
- LUMBAL REGION
- SUBOCCIPITAL : CISTERNA MAGNA REGION
- VENTRICEL REGION
LUMBAL PUNCTURE
L3/L4
DANGER ! :
- INTRACRANIAL PRESSURE
- INFECTION
- PAIN

DISCARDED
EXAMINATION
MICROSCOPIC
MACROSCOPIC
CHEMISTRY
BACTERIOLOGY
IMMUNOLOGY/
SEROLOGY
TO PREVENT
SPONTANT
COAGULANT
20% Na Citrat
0.01 ml/ml LC
STERIL
TUBE
A. MACROSCOPIC EXAMINATION :

1. COLOUR :
- NORMAL : CLEAR AND COLOURLESS
- ABNORMAL :
a. RED = TRAUMA OF PUNCTURE
= SUBARACHNOIDAL BLEEDING
b. BROWNISH COLOUR = OLD BLEEDING HAEMOLYSIS
c. YELLOWISH COLOUR = OLD BLEEDING
ORANGE-YELLOWISH = ICTERUS HIGH BILIRUBIN CONTENT, HIGH
PROTEIN CONTENT/HIGH LIPIDE
d. GRAYISH COLOUR = HIGH LEUKOCYTE COUNT

2. TURBIDITY :
CAUSED BY :
a.. ERYTHROCYTE CONTENT
b. INFLAMMATION CELLS (LEUKOCYTE, EPHITEL CELL, BACTERIA)

NOTES : HIGH LEUKOCYTE NOT ALWAYS MAKE THE LIQUID IS TURBID,
LIKE COUNT :
- MENINGITIS SEROSA (TBC. LUES, VIRUS)
- TABES DORSALES
- POLIO MYELITIS
RESULT : CLEAR, RATHER TURBID, TURBID, MORE TURBID

3. SEDIMENT : N = NEGATIVE AFTER SENTRIFUGATION

4. CLOTTING : N = NEGATIVE
AN = POSITIVE : FINE, THREADS, FILAMENT
WHEN FIBRINOGEN POSITIVE = HIGH ALBUMIN,
GLOBULIN HIGH
- IN THE CASE OF TUBERCOLUSIS : FINE WEB OR PELLICLE AFTER STAND
FOR 12 HOURS
- IN THE CASE OF MENINGITIS PURULENTA : THAT COAGULATION IS
COARSE AND MASSIVE
- EN MASSE COAGULATION MAY BE ABLE TO FIND IN :
FROINS SYNDROME : - XANTHOCHROM
- PROTEIN HIGH
- PLEIOSITOSIS LYMPHOCYTE

5. PRESSURE : N = 70 - 200 mm H
2
O
B. MICROSCOPIC EXAMINATION OF CSF
ERYTHROCYTE AND LEUKOCYTE COUNT SHOULD BE PERFORMED
WITHIN 30 AFTER THE SPECIMENT IS OBTAINED
1. CSF LEUKOCYTE COUNT

MATERIAL :
1. HEMOCYTOMETER FUCHS-ROSENTHAL= AREA 4 x 4 = 16 mm
2

DEPTH = 0.2 mm
2. CONCENTRATE TURK SOLUTION
METHODS : - DRAW TURK = 1.0
- DRAW CSF = 11.0
DILUTION
9
10
- MIX GENTLY, DISCARDED THE FIRST
3 DROPS
- FILL THE HAEMOCYTOMETER
- COUNT THE LEUKOCYTE WHICH ITS
LIES IN WHOLE HEMOCYTOMETER
FIELD, IF THE CALCULATED ARE N CELLS
LEUKOCYTE COUNT :
N
16
10
2
10
9
50 N
144
N
3
X X = =
NORMAL : 0 - 5 / mm
3
CONSIST OF LYMPHOCYTES
( < 5TH : 0 - 20 / mm
3
)
ABNORMAL : > 10 / mm
3

M. PURULENTA : 500 - 20.000 mm
3

M. SEROSA : SPI 200 / mm
3

ERYTHROCYTE : NORMALLY ABSCENT
WHEN USED AN IMPROVED NAUBAUER
CALCULATION :
VOL = 9 X = mm
3
= N. CELLS
1
10
9
10
N X X = = N
10
9
10
9
100
81
5
4
CELL COUNT :
CSF = 0.05 ml
TURK = 0.95 ml
DILUTION = 1/20
FUCH ROSENTHAL HEMOCYTOMETER
2. COUNT THE CELLS IN 5 mm2 (USING SQUARE 1,4,7,13,16)
WHEN = N. CELL
TOTAL CELLS = 20 X N
THE OTHER COUNTING METHOD :
1. MIX
2. DIFFERENTIAL COUNTING
CFS CENTRIFUGE 1500 - 2000 RPM / 10
SEDIMENT STAINED BY GIEMSA /WRIGHT
DIFFERENT COUNT : PMN / LYMPHOCYTE
HIGH LYMPHOCYTE = MILD - CHRONIC INFLAMATION
= TBC, LUES, VIRUS
HIGH PMN = - ACUTE INFLAMATION
- BACTERIAL MENINGITIS
= @ MENINGITIS PURULENTA
@ BRAIN ABSCESS
@ EXTRADURAL ABSCESS
3. CHEMICAL EXAMINATION OF CFS
A. PROTEIN
NORMAL : 15 - 45 mg / dl
PREMATURE INFANT : 400 mg / dl
1 YEAR INFANT : 30 - 100 mg /dl
ELDERLY : 30 - 60 mg / dl
A.1. QUALITATIVE TEST FOR PROTEIN
FOAM TEST PANDYS TEST
NONNES TEST
(NONNE APELT)
ROSS JONES
SHAKE
HARDLY
N : 1 - 2 FOAM
DISAPPEARED
AN : 5 REMAIN

R/
PHENOL LIQUEFACTUM 10ml
AQUADEST 90 ml
RESULT :
+ : WHITE CLOUD
- : NO WHITE CLOUD OR
SLIGHT CLOUDNESS

GLOBULIN
CFS
REAGENT :
1 ml REAGENT
1 DROP CFS
CFS aa
1/2 - 1 ml REAGENT
R/ AMMON. SULFAT
CONCENTRATE IN WATER
STAND FOR 3
LOOK RING
+ : APPEAR WITHIN 3
SHAKE, DISAPPEAR
++ : SHAKE, OPALESCENT
+++ : SHAKE, CLOUDLY
++++ : SHAKE, THICK CLOUDY
ALBUMIN
MEAN :
HIGH PROTEIN CONTENT
A.2. TEST FOR QUANTITATIVE PROTEIN
PHOTOCOLORIMETRIC
TURBIDIMETRIC
ELECTROFORESA : PROTEIN FRACTION
N : PROTEIN : L.P. : 15 - 40 mg /dL
CYS. MAG. : 10 - 25 mg /dL
VENTR. : 5 - 15 mg /dL
CONSIST OF ALBUMIN
AN : HIGH GLOBULIN CONTENT, FIBRINOGEN

HIGH PROTEIN CONTENT : M. PURULENTA
M. TBC
BLOCK
SUBARACH BLEEDING
B. GLUCOSE
N : 50 - 80 mg / dL ( 1/2 BLOOD GLUCOSE)

M. PURULENTA
M. TBC
GLUCOSE DECREASEED
C. CHLORIDE ( NaCl )

N : 720 - 750 mg /dL (120 - 130 mg/dL)
BLOOD 550 - 620 mg /dL

M. PURULENTA DECREASED : ( < 680 mg / dL)
M. TBC MORE DECREASED : ( < 600 mg / dL)
DISCHARGE FROM : VAGINA, URETHRA, PROSTATE
: INJURY
> MACROSCOPIC EXAMINATION
> MICROSCOPIC EXAMINATION
SPECIMENT. SMEAR GRAM / BLUE METYLINE
DISCHARGE OF (PUS) :
W URETHRA
W VAGINA
W CERVIX










W PROSTATIC SECRET COLLECTED BY MASSAGE
- GO
- CANDIDA ALBICANS KOH
- TRICHOMONAS VAGINALIS


1 GTT SPECIMENT + 95% SALINE


MICROSCOPE
DETECT FLAGELLA
(MOTILITY)

SMEAR PREPARATION
GRAM STAINING
CULTURE
CEMEN ANALYSIS
THE CEMEN CONSIST OF : - SPERMATOZOA
- SECRETION OF GLANDULA DUCTUS :
@ D. DEFERENS. / D. GENITALIA
@ GLANDULA V. SEMINALIS
@ GLANDULA PROSTATE
@ GLANDULA COWPERI (GL. BULBOURETHR)
CHEMISTRY : SPERM : NUCLEO PROTEIN
THE FLUID OF CEMEN :
- PHOSPHATASE ACID
- CITRIC ACID
- PHOSPHORILCHOLIN
- CHOLIN
- FIBRINOLISIN
- PROSTAGLANDIN
- SPERMIN
- FRUCTOSE
FRUCTOSE :
- THE SOURCES OF ENERGY FOR SPERM
- ESSENSIALE FOR METABOLISM & MOTILITY OF SPERM
- ITS FORMATION IS INFLUENCED BY TESTOSTERON
- IN CONNECTION TIGHTLY WITH INFERTILITY
- BALANCE RECIPROCAL WITH THE COUNT OF SPERM
INDICATION FOR CEMEN EXAMINATION:

= MALE INFERTILITY
EXAMINATION : - MACROSCOPIC
- MICROSCOPIC
- CHEMISTRY

COLLECTION OF SPECIMEN :

PREPARATION SEXUALLY ABSTINENTIA 3 - 5 DAYS
COLLECTION OF CEMEN : MASTURBATION


ALL THE SPECIMEN OF CEMEN MUST BE COLLECTED IN A CLEAN
AND LARGE MOUTH CONTAINER
MACROSCOPIC EXAMINATION :

COLOUR : WHITE OR GREY WHITE
SMELL : MUSTY OR ACRUD ODOUR
VOLUME : 2 - 5 ml
AFTER 10 - 20 MINUTE LIQUIFY TO FORM A
TRANSLUCENT, TURBID, VISCOUS FLUID
VISCOCITY : THE SPECIMEN OF NORMAL VISCOCITY CAN BE
POURED DROP BY DROP.
INCREASED VISCOCITY IS OF SIGNIFICANT IF
SPERM MOTILITY IS THERE BY COMPROMIZED AND
HAS BEEN SHOWN TO BE ASSOCIATED WITH
POOR INVATION OF THE SERVICAL MUCUS AND
MAY BE DEFECT IN FERTIL COUPLE
MICROSCOPIC EXAMINATION :
1. SPERM COUNTS
MATERIAL : - HEMOCYTOMETER
- DILUTION SOLUTION
R/ SODIUM BICARBONATE 5 gr
FORMALIN 1 ml
AQUADEST 100 ml

METHODS :
- DRAW THE CEMEN TO THE MARK = 0.5
- DRAW THE SOLUTION TO THE MARK = 11.0
DILUTION IS 1 : 20
- CHARGE TO THE HEMOCYTOMETER CHAMBER
- STAND FOR 2 MINUTE
- COUNT SPERM IN 4 LARGE SQUARE ( 4 mm
2
) = N. CELLS
- RESULT = SPERM / ml = N X 50.000
NORMAL : 70.000.000 - 300.000.000 / ml
2. MOTILITY OF SPERM
SMALL DROP
37
0
C
CEMEN
OBSERVE 200 SPERM
- SCAN SEVERAL FIELD 40X
TOTAL + 200 SPERM
- FOCUS THE ENTIRE DEPTH
NON MOTILE SPERM
- RECORD % OF MOTILITY SPERM
N : 70 - 80 % ACTIVE ( WITHIN 1 HOUR)
IMMATURE SPERMATOZON
(SPERMATID)
3. MORPHOLOGY
- PREPARE THE SMEAR
- STAIN WITH : GIEMSA, FUCHSIN. ETC
- + 200 SP EXAMINE 200X

HEAD
NECK
TAIL
3 - 6 m m
50 - 70 m m
ABNORMAL :
PIN HEAD
GIANT HEAD
ACUTE TAPERING FORM
AMORPHOUS FORM
DOUBLE HEAD
DOUBLE TAIL
CONSTRICTED HEAD
NOTES : TAIL ON ALL FORMS ARE DISPROPORTIONAL SHORT
AGGREGATION
HEAD TO HEAD
HEAD TO TAIL
TAIL TO TAIL
NECK TO NECK
HEAD TO NECK
NECK TO TAIL
MANAGEMENT OF ROUTINE AND SIMPLE
CLINICAL LABORATORY
CLINICAL PATHOLOGY : LABORATORY
ROUTINE & SIMPLE
CLINICAL LABORATORY

- URINALYSIS
- HAEMATOLOGY
- FAECES
- CSF/BODY FLUIDS
CLINICAL PATHOLOGY
DEPARTMENT OF :
1. CLINICAL HAEMATOLOGY
2. CLINICAL BIOCHEMISTRY
3. CLINICAL ROUTINE EXAM
4. CLINICAL MICROBIOLOGY
5. CLINICAL IMMUNOLOGY
6. BLOOD BANK
7. EMERGENCY
MANAGEMENT
SUCCES
TO
REACH
AMBISION
AND
TARGET
PERFORMANCE :
EFFECTION
&
EFFICIENT
CLIN. PATH LAB.
PHYSICOLOGY
BIOLOGY
BASIC
MEDICAL
SCIENCE
MEDICAL
PRACTISE
DESIGN OF MANAGEMENT
FROM ROBBIN
FINANCIAL

PHYSICAL

HUMAN
ORGANIZING

PLANNING

DIRECTING

CONTROLLING

STAFFING
SATISFACTORY
PERFORMANCE

PRODUCT

SELF SERVING
BEHAVIORS
POLITICS
POWERS
CONFLICT
RESOLUTION
I
N
P
U
T
POLITICS
POWERS
CONFLICT
RESOLUTION
ORGANIZATION ( CLIN. LAB. ROUTINE/SIMPLE )

STRUCTURE = RELATIONSHIP OF FRAMEWORK
PROCESSES = INTERACTION
ELEMEN :
- WORKING PLACES
- STAFF = JOB DISCHARGE
- JOB = WHICH MUST BE SOLVED
- MANAGER = THEY HAVE TO MANAGE THE JOB FROM
THE OTHERS
- EXECUTIVE = ALL OF THEM ( IN MODERN ORGANIZATION)
WHO WHICH BY VIRTUE OF POSITION AND KNOWLEDGE,
RESPONSIBLE TO CONTRIBUTE A JOB WHICH SUPPORT
ORGANIZATION CAPACITY FOR WORK AND TO GET RESULT
O
U
T
P
U
T
KEY PERSON : CONSIST OF
1. LABORATORY DIRECTOR
2. MANAGER
3. SUPERVISOR
4. TECHNOLOGIST

WHOSE HAVE :
1. MOTIVATION
2. VISION
3. CAPABLE OF DECISION MAKING
4. HEALTH = WELL-BALANCE : PHYSICAL, EMOTIONAL,
SPIRIT AGAINTS TENSION, FRUSTRATION, STRAIN
5. HUMILITY = NECESSARY FOR HELP FROM THE OTHER PEOPLE
EFFICIENCY OPERATION OF CLINICAL LABORATORY AND
DELIVERY OF MEDICAL LAB. SERVICES TO THE PATIENTS
OR PHYSICIANS
NECESSARY COMPLEX COOPERATION FROM :
- MEDICAL, SCIENTIFIC AND TECHNICAL DEPARTMENT
- THE SOURCES IN FORM OF FACILITY PERSONAL,
LABORATORY EQUIPMENT, PROCESSING DATA, AND
EQUIPMENTS
- SKILL IN MANAGEMENT, ORGANIZATION AND
COMMUNICATION
REGULATION, COMPETITION, MALPRACTISE, ETC
BASIC SCIENCE
RESEARCH
EDUCATION
MEDICAL PRACTICE
DIAGNOSIS
THERAPY
PROGNOSIS
QUALITY ASSURANCE
REAGENTS
EQUIPMENTS
COMPUTER SCIENCE
MANAGEMENT TECHNIQUES
INDUSTRY
BRIDGING