Drs. H. Endang Samaun W, dr.

, SpPK-K
 A. DEFINITION OF ROUTINE CLIN. LAB
 Hb.
BLOOD.  LEUKO. CONT.
I. “ROUTINE”  DIFF. COUNT.
CLIN LAB B. EXAM.  B.S.R.
EXAM  MACR/MICR
URINE.
 PROT/RED.
FAECES.  MACR/MICR.

C. TO ESTIMATE A LIMITATION

  ERY,THROMBO,
RETIC
 PCV, MCV, MCHC
BLOOD.  P. SLIDE/PARAS
 BLOOD GROUP
CLIN  RL/BT/CT/CR
PATH
 UROB.  PREG T.
 BIL  SED
URINE.
II. “SIMPLE” A. EXAM  BENZ  ACID
CLIN LAB  ESBACH  SP. GR
EXAM
FAECES.
SPUTUM.
CEREBRO SPINAL FLUID
TRANSUDATE / EXUDATE
SECRETE : VAG. / UR. / INJ.
CEMEN ANALYSIS.

B. TO ESTIMATE A LIMITATION

III. MANAGEMENT OF ROUTINE & SIMPLE CLIN. LAB

URINE SPECIMENS
1. THE MORNING’S 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 AT 06.00
GETS UP THE OTHER DAY
URINATES
(AT 06.00)

DISCARDED URINE PASSED DURING

THE REST OF THE DAY
 (MORNING)
URINE
(NEW)

SHAKE

MACROSCOPIC

COLOUR
SMELL
CLOUDY
ACIDITY SEDIMENT
SPEC. GRAF
SUPERNATANT

MICROSCOPIC CHEMIC

ERYTHROCYTE ALBUMIN
LEUKOCYTE GLUCOSE
EPHITEL UROBILIN
CRYSTAL BILIRUBIN
ROUTINE CAST KETOBODY
SIMPLE BENZIDIN
1. MACROSCOPIC EXAMINATION OF URINE

A. COLOUR

 LIGHT YELLOW (TEA) NORMAL

 DARK YELLOW BILIRUBIN (?)

FOAM TEST

YELLOW (OBVIOUS)
SHAKE = F. T +
(HARDLY) FOAM > 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
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 H2S

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 CL2 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.)
 S.G. IS DEPEND ON THE TOTAL OF SOLUTE SUBSTANCES
 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)
1.000
1.020 CORECTION

1.040
 TEMP. : EVERY 30 C > 150 C : + 0.001
40 C > 170 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
CLINICAL VALUE OF URINE SPECIFIC GRAVITY.
GRAVITY
 TO HELP THE DIAGNOSIS GLUCOSURI OF COMATOUS PATIENT
 FISHBERG CONCENTRATION TEST FOR RENAL FUNCTION

METHODS
 PATIENT DON’T 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
2. MICROSCOPIC EXAMINATION OF URINE

NEW URINE < 6 HOURS

CENTRIFUGE AT 1500 RPM / 5 MINUTES

SEDIMENT COVER WITH

COVER GLASS
DROP

SLIDE

MICROSCOPE OBJECTIVE 40 X
EYEPIECE 10 X
CONDENSOR

EXAMINATION ! !

ERITHROCYTE / LOW POWER

LEUKOCYTE / HIGH POWER

ORGANIC
SEDIMENT
CAST / LOW POWER

EPHITEL CELL

CRYSTAL
ANORGANIC
SEDIMENT
ERITHROCYTE

MORPHOLOGIC :

A. NORMAL (NEW URINE) :

- ROUND,  + 7  (EQUALLY)
- YELLOWISH

B. CRENATED (HIGH SPECIFIC GRAVITY URINE)

- DARKER AT THE EDGES
- SPIKY EDGES
- THE FLUID OUT OF THE CELLS

CLINICAL VALUE :
NORMALLY, THERE ARE NO RED CELLS IN URINE

ERY +URINE : - BLEEDING (CANCER OF REN, PYELUM)

(HEMATURI) - TRAUMA (CALCULUS, CRYSTAL)
- INFLAMMATION (TBC, GNA)
FAR ADVANCED EXAMINATION

NOTES :

ERY +: : REPEATED BY MIDSTREAM URINE

OR CATHETER URINE

NORMAL : 0 - 1 / LPB
 LEUCOCYT (WHITE CELLS)

MORPHOLOGIC :

 CLEAR GRANULAR DISKS

 ROUND ;  + 11 (1.5 - 2 E)
 THE EDGES NOT CLEAR
E  CELLS SURFACES ARE GRANULAR
E
 IN NEW URINE AMOEBOID MOVEMENT
IN NEW ALKALIST CLUMPS
 IN ALWAYS FOUND 2 - 6 LEUCOCYT OR MORE / LOW POWER

- CATHETER URINE >6 / HIGH POWER = PATOLOGIC

- MEN

THE CLINICAL VALUE : N : 0 - 6 / LOW POWER

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
CAUDATUS
EPITHELIAL CELLS ROUND EPITHELIAL
EPITHEALIAL CELLS
(VAGINA) (TUBULUS)
(PELVIS R.)
(URETHRAE DIST.)

 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
APPEARENCE OF CRYSTAL

- Ca OXALATE :

 SIZE VARIABLE
 CLEAR
 MORPH. LIKE ENVELOPE

CLEAR
- PHOSPHAT :

AMMORPHUS
TRIPLE CALCIUM PHOSPHAT
PHOSPHAT PHOSPHAT

- CARBONATE : CALSIUM
CARBONATE

- URIC ACID :

COLOUR
IS BROWN

- URATE :
Na - URATE

NH4 URATE

AMMORPHUS
URATE
CRYSTAL

NORMAL CRYSTALINE DEPOSITE

1. CALCIUM OXALATE (ACID URINE)

A. SHAPE LIKE ENVELOPE
SIZE 10 - 20  m
B. SHAPE LIKE PEANUTS
SIZE + 50  m
COLOUR : CLEAR / TRANSPARANT
( COLOURLESS )

2. URIC ACID (ACID URINE)

SHAPE VARIES (SQUARE, DIAMOND SHAPE, CUBICAL/
ROSE SHAPE)
SIZE 30 - 150  m
COLOUR YELLOW - BROWNISH RED

3. TRIPLE PHOSPHATES ( NEUTRAL / ALKALINE URINE )

SHAPE : A. RECTANGULAR
B. LIKE A TERM LEAF / STAR
SIZE 30 - 150  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

CERTAIN
 PRINCIPLE : PROTEIN CLOUDY
pH
 CONDITION OF URINE : - ACID AND CLEAR
 CHARACTER OF EXAMINATION : QUALITATIVE /
SENSITIVITY 5 - 10 mg /dL

A. EXTON TEST FOR PROTEIN URINE

REAGEN : SULFOSALISILIC ACID : 25 GR. 50

URINE NA2SO4 : 100 GR. 88
AQUADEST : 500 CC. 1000

SENTRIFUS  CLEAR = PROTEIN -

(NO WHITE PRECIPITATE)
SHAKE  TURBID = EXTON +
REAGENS : 2.5 ml
URINE : 2.5 ml FOLLOWED BY :
1. BANG TEST
COMPARE WITH A TUBE OF UNTREATED URINE 2. ACETO PRECIPITABLE
AGAINTS A BLACK BACKROUND SUBSTANCE TEST

EXTON + : - PROTEIN
- PROTEOSEN
- BENCE JONES PROTEIN
- URIC ACID & OTHERS
 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

RESULT : SYM PROT

BOL (mg %)
CLEAR - 0
SLIGHT TURBID + + 10
BOILED
READ TURBID WITHOUT ++ 10 - 50
REAGEN 0.5 ml GRANULE
URINE 5 ml
TURBID WITH ++
++ 50 - 200
GRANULE

1000 C
TURBID WITH +++
+++ 200 - 500
10‘
FRAGMENT
WATER BATCH
CLOTTING ++++ > 500

IF THE AMOUNT OF ALBUMIN 3000 mg % CLOTTING

BOILED

- PROTEIN : -
EXTON
- PROTEOSEN
+ BANG
PROTEIN
+
BENCE JONES PROT.

TURBID WITHOUT BOILING

ACETOPRECIPITABLE
 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)

PUT THE RUBBER OR CORKY SHUTTER
PIPET REAGEN READ :
THE HEIGHT
SHAKE OF WHITE
PRECIPITATION

PIPET URINE MOVING 5

THE TUBE
(10 X)
ALBUMINOMETER
5 GR / L / 24 HOURS
PLACE :
- IN ROOM TEMP.
- FOR 24 HOURS
- AVOID TO LIGHT AND
UPRIGHT POSITION
 TEST FOR “ACETOPRECIPITABLE” SUBSTANCE

+ WATER FULL

30% ACETIC ACID

SOLUTION

gtt
1- 2 t
URINE 10 ml PUT gt
4
2-
t
Pu 0 gtt

Urine Urine Urine

2 ml 2 ml 2 ml

I II III

RESULT : SECOND TUBE : - CLEAR : ACETOPRECIPITABLE

- TURBID : MUCIN

CLEAR TURBID
 E. EXAMINATION OF “BENCE JONES PROTEIN”
 CHARACTERISTIC OF BENCE JONES EXAM. :
- PRECIPITATE : 440 - 500 C
- SOLUBLE AT TEMPERATURE > 950 C
 SCREENING TEST :
ADD HCL CONCENTRATE GENTLY,
SO THAT LIMITATION BETWEEN
HCL CONCENTRATE URINE AND HCL

- = B.J.P. -

PRECIPITATION
RING + = - GLOBULIN
URINE - PROTEOSA
5 ml
-B-S-P

 CONFIRMATION TEST :

1 DROP ACETIC ACID SOLUTE 2%

pH : 5.5

10 ml ADD Na Cl CONCENTRATE 2 ml
URINE

READ THE TEMPERATUR

100 C0 IN WHICH ITS PRECIPITATION
OCCUR
WATER BATCH
+ : - MULTIPLE MYELOMA
- CLL, CML
- AMYLOID DESEASE
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 MIX AND ADD
SODIUM CARBONATE 135 GR. 100 500 ml WATER
DISTILLED WATER 350 GR.
SIGN GLUCOSE
COPPER SULFATE 85 GR 17.3 gr/dl
WATER 50 ml 100
- METHOD :
BLUE - 14
0-0.1

TRACE GREEN + 28
WITH YELLOW

BOIL LEAD TO COOL/

ROOM TEMP.,
BENEDICT 5 ml
READ THE RESULT
YELLOW ++ 56
URINE 8 GTT

1000 C
WATER BATCH
BROWN +++ 83

5‘

ORANGE ++++ 111

TO BRICK RED
 ALIMENTARY
A. DIABETES
MELLITUS HYPERTHYROID

CUSHING
B. SECONDARY
GLUCOSE SYNDR.
GLUCOSURIA

ACROMEGALIA
C. GLUCOSURIA
A. SUGAR RENAL INTRACRANIAL
HYPERTENTION

A. LACTOSE
THE OTHER
SUGAR B. FRUCTOSE

C. PENTOSE
REDUCTION
+ D. GALACTOSE

SALICYL
DRUGS ANTIPYRIN
UROTROPIN
SANTONIN
B. THE OTHER
SUBSTANCES URIC ACID
THE OTHER
SUBSTANCES KREATININ

INDIKAN
 BILIRUBIN METABOLISM

ERI
RUPTURE
HAEMOGLOBINE

HAEM + GLOBIN + Fe

BILIVERDINE

BILIRUBIN  PREHEPATIC BILIRUBIN

 INDIRECT BILIRUBIN
- WATER IN SOLUBLE
- ALCOHOL SOLUBLE
CONJUGATION
WITH GLUCURO-
NICACID
 DIRECT BILIRUBIN
CONJUGATED - WATER SOLUBLE
REN BIL.

UROBILINOGEN
STERCOBILINOGEN
UROBILIN

FAECES
(STERCOBILIN)
 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 : EHRLICH’S REAGENT

R/ : - P. DIMETYL AMINO : 2 gr
BENZALDEHYDE
- 37% HCl : 50 ml
- AQUADEST : 100 ml

SHAKE + DEEP RED COLOUR

REAGENT 10 ml
URINE 5 ml

- A FINE PINK COLOUR

NORMAL : 2 mg / 24 HOURS
 B. DETECTION OF UROBILIN IN URINE
DISTURBANCE WITH EHRLICH
OXYD.
PRINCIPLE : UROBILINOGEN IOD 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

METHOD :

R
E R PE
A
LT P
FI ER
LT
FI
BY LO
OK
7.5 ml REAGENT
SHAKE FO
2 GTT LUGOL R

5 ml URINE
OU UR
ND
GR OLO

A PRECIPITATE FORMS
CK C
B A ACK

STRONG GREEN FLUORESCEN = +

BL

LIGHT GREEN FLUORESCEN = NORMAL

UROBILINURI :
- HEMOLISES = A.N HEMOLITICS
- HEPATITIS ETC.
 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. FOUCHET’S SOLUTION
R/ TCA 25 gr
10% FeCl3 10 ml
AQUADEST 100 ml
2. 10% SOLUTION OF CACl3
3. SOLUTION OF Na2CO3 CONCENTRATE
OKS
PRINCIPLE : BILIRUBIN FeCl3 BILIVERDIN (GREEN)
METHOD :

R
T ER PE
L A
FI R P
E
LT
FI
Na2CO3 20 GTT SHAKE
CaCl2 10 GTT
U 5 ml

A PRECIPITATE FORMS
+ : COLOUR : GREEN BLUE

RESULTS : - = NO COLOUR CHANGE

+ = THE PRECIPITATE TURNS GREEN / BLUE
B. HARRISON/HAWKINSON MODIFICATION
TO DETECT BILIRUBIN IN URINE
LARGE
FILTER PAPER

RINSE

10% BaCl2 SOLUTION TEST

I. URINE II. FOUCHET SOLUTION

TEST :
1. PUT ONE DROP OF URINE
2. PUT ONE DROP OF FOUCHET SOL.

ALLOWED TO DRY
AND MAKE LITTLE PIECE OF += GREEN TO BLUE COLOUR
FILTER PAPER BY CUTTING

NOTES :
1. FOUCHET TEST :
- VERY SENSITIVE
- DIAGNOSIS CAN MADE EARLY
2. BILIRUBIN + BY :
A. OBSTRUCTION OF DUCTUS BILIARIS :
- CALCULUS
- INFLAMATION
- TUMOR, ETC
B. HEPATITIS
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 +++ / ++++ KETOSIS DIABETICUM

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
A. ROTHERA METHOD
PRINCIPLE :
SUBSTANCE WITH
- ACETON
- ACETOACETIC ACID
+ Na. NITROPRUSIDA VIOLET COLOUR

MATERIAL : A. Na. NITROPRUSIDA 5 gr

AMMONIUM SULFATE 200 gr
B. AMMONIUM HYDROXIDE CONCENTRATE
METHOD:
NH4OH
CONCENTRATE

VIOLET COLOUR RING

+ BRIGHT VIOLET
MAKE COMPLETE
SOLUTION READ RESULT ++ DARK VIOLET
WITHIN 5‘ +++ DARK AND WIDE
URINE 5 ml
REAGENT 1 gr 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 + FeCl3 WINE RED SUBST.
THIS TEST IS LESS SENSITIVE THAN ROTHERA METHOD
 REAGENT : 10% FeCl3 SOLUTION

METHOD :

2 DROPS
FeCl3 SOLUTION

ASETOACETIC ACID

FALS POSITIVE :
SHAKE
+ - FENOL
(RED) - SALICYLAT
URINE 5 ml - ANTIPYRIN
- Na. CARBONATE

MUST BE DONE

ROTHERA OR SECOND GERHARDT

METHODS METHODS
STILL+ :
THE OTHER SUBSTANCES

AQ. 5 ml
FeCl3

COOK

U : 5 ml
U : 5 ml

CHANGE TO - :
KETONE SUBSTANCE
 DETECTION OF OCCULT BLOOD IN URINE

IF : HEMATURIA SEDIMENT EXAM. ERI +

Hb. URIA SEDIMENT EXAM ERI -
BENZIDINE TEST
PRINCIPLE :
BLOOD PEROKSIDASE ACTIVITY

H2O2 H2O + On BENZIDINE OXIDATION

METHOD :

PREPARE :

1. BENZIDINE
SOLUTION
3. H202 SOLUTION

5ml ACETIC ACID TEST

GLACIAL

1 gr BENZIDINE
l
3 ml 1m

2. URINE
GREE
N +
l
BL
COOK m UE
2
+
(H
READ WITHIN AR
5‘ D )
URINE MAKE COOL

 THIS TEST IS VERY SENSITIVE - URINE MUST BE HEATED

 GIVE + RESULT BY OXYDASE - EQUIPMENT CLEAN
FROM LUEKOCYTE
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

COMPARE
READ : 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

METHOD :

1. DROP 1 GTT URINE

2. DROP 1 GTT ANTI-HCG

MIXED ( + 1‘)

3. DROP 1 GTT LATEX

1 MINUTE
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 BLOOD CELLS

(55%) (45%)
 ERYTHROCYTES
 LUEKOCYTES
 THROMBOCYTES
WATER SUBSTANCES (PLATELETS)
(90%) (10%)
 PROTEIN :
- ALBUMIN
- GLOBULIN
- FIBRINOGEN
 CARBOHYDRATE
 LIPID
 VITAMIN
 MINERAL
 ENZYM
 BLOOD COLLECTING PROCEDURES

I. CAPILLARY BLOOD

A. LOCATION : 3
4
ADULT : THE TIPS OF FINGER
BABY : HEEL OR BIG TOE X
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 1st DROP OF
BLOOD WITH DRY COTTON
WOOL
 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 TORNIQUET
& NEEDLE
- 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) A.C A.C

METHOD :
 READ CAREFULLY THE PATIENT’S 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
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

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 B
WITHOUT WITH
ANTICOAGULANT ANTICOAGULANT

SOLIDIFIES WITHIN 5 - 20‘ CLOTTING IS PREVENTED

FORMING REMAIN FLUID

PLASMA

SERUM WBC

BLOOD CLOT RBC

THE DIFFERENT :
- FIBRINOGEN +
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
 40 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 LEED’S TEST

13. BLEEDING TIME

14. COAGULATION TIME

15. CLOT RETRACTION

 HAEMOGLOBIN ESTIMATION

METHODS : 1. TALLQVIST COLOURIMETRIC

2. SAHLI
3. PHOTOMETRIC

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 IT’S MADE FROM PAPER
- IT SHOULD BE CORRECTED BY
THE MORE EXACTLY METHOD

100% 15.8 gr %
 2. THE METHOD OF SAHLI
a b
PRINCIPLE : Hb + HCl HEMATINE ACID (COLOUR BROWN)
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: 2
3

1
5 6
4
 NOTES :

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 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
 3. PHOTOMETRIC METHOD

CYANMETHEMOGLOBIN

K. CYANIDA (KCN) CYAN-MET Hb

PRINCIPLE : BLOOD DILUTED K. FERRICYANIDA (STABIL )
(Hi CN)
DRABKIN DILUTING FLUID:
R/ NaHCO3 : 1.0 gr
(EXAM IN SPECTRO)
KCN : 0.05 gr
K3Fe(CN)6 : 0.20 gr
AQUA : 1000 ml
MATERIAL :
 SPECTROPHOTOMETER
 PIPETS & TUBES
 DRABKIN DILUTING FLUID
 CYAN MET Hb STANDARD

METHOD :

INTO A TUBE, PIPET : MIX THE CONTENT

5 ml DRABKIN DILUTING FLUID OF THE TUBE,
0.02 ml BLOOD LEAVE FOR 5 ‘

READ PHOTOMETRIC (520 nm)

CALCULATION :
ABSORBANCE OF SAMPLE
X Hb STANDARD ( 15 g/dL)
ABSORBANCE OF STANDARD
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
 METHODS OF :

LEUKOCYTE AND ERYTHROCYTE

COUNT

MATERIAL :

CONSIST OF
 FOR COUNTING :
LEUKOCYTE ERYTHROCYTE

1. THOMA’S PIPET


2. COUNTING CHAMBER

.. - HAYEM
3. DILUTING FLUID TURK - FORMOL CITRATE

4. HEMOLANCET
COTTON + 70% ETHANOL


COUNTING CHAMBER = IMPROVED
.. NEUBAUER
BURKER , ETC.
A B C

COLOUR BAND
&
NEWTON’S 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 NEWTON’S 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

0.05 mm
1 mm 1 mm

X = LEUKOCYTE COUNT SQUARES

1 1 1
X X mm3
5 5 10
= 25 SQUARES
A = ERYTHROCYTE COUNT SQUARES

1 1 1
X X mm3
20 20 10
= 80 SQUARES
1 1 1 = 1 mm3 = N
VOL = 80 X X X
20 20 10 50
E = N . 50 . 200 = N X 10.000/ mm 3
1 2 3
B = 1 1 1
VOL (3 X 0.05 X 0.1 mm3) = 0.015 mm3 X X mm3
5 20 10
3 SQR = 3 X 0.015 = 0.045 mm3 N
S

1 200 = 20 SQUARES
E=N X 200 = N X
0.045 0.045 1 1 1 = 1 mm3 = N
VOL = 20 X X X
= N . 4.444 5 20 10 50
E = N . 50 . 200 = N X 10.000/ mm 3
IMPROVED NEUBAUER

LEUKOCYTES COUNT :
4 CORNER SQUARES ( L ) = 1 X 1 X 0.1 mm3
ERYTHROCYTES COUNT :
1 1
5 SQUARES ( E ) X X 0.1 mm3
5 5
1 1 1
80 SQUARES : X X mm3
20 20 10
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 THOMA’S 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 IMPROVED NEUBAUER
CHAMBER COUNTING CHAMBER LARGE CORNER SQUARES
1
4 1X1X mm3 = N. CELLS
10
4 mm3 = N. CELLS
10
1 mm3 = N X 4 X 20
10
200
=NX CELLS
4

1 1 1
25 SQR : X X mm3 N. CELLS
S

5 5 10
NORMAL :
1
mm3 N. CELLS ADULT : 4.000 - 10.000 mm3
S

10
INFANT NEW BORN : 10.000 - 12.000 mm3
1 mm3 N X 10 X 20
S

INFANT 1-6 MOUTH : 4.000 - 15.000 mm3

N X 200 CELLS CHILDREN 4 - 7 YEAR : 4.000 - 11.000 mm3
S

8 - 12 YEAR : 4.000 - 10.000 mm3

> N = LEUKOCYTOSIS BACTERIAL INFECTION
EXCEPT : S. TYPHOSA
< N = LEUKOPENIA INFECTION : S. TYPHOSA
MALARIA
- 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 LEUKOCYTE = 16 X 109 /L
50 X 16 = 5.3 X 109 /L
FORMULATION N =
100 + 50
LEUKOCYTE = 16 - 5.3 = 10.7 X 109 /L
 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

3
6

5
HAYEM
SOLUTION
COUNT BY MICROSCOPE
CONDENSOR : DOWN WARD
OBJECTIVE : 10 X /40X

CALCULATION

..
BURKER COUNTING
CHAMBER
IMPROVED NEUBAUER
COUNTING CHAMBER

COUNT IN THE RED CELLS IN COUNT IN THE RED CELLS IN

1 1 1 mm3
1 1 1 5 SQUARES X X
A. 80 SQUARES X X mm3 5 5 10
20 20 10
= N. CELLS = N.CELLS
1 1 1 1 1 1 1 1
VOL = 80 X X X = mm3 VOL = 5 X X X = mm3
20 20 10 50 5 5 10 50
1 1 1 mm3 = 50 X 200 X N. CELLS
mm3 BLOOD
50 200 = N. CELLS (WHOLE BLOOD) = 10.000 X N. CELLS
1 mm3 = 50 X 200 X N. CELLS
(WHOLE BLOOD) = 10.000 X N. CELLS

1 1 1
B. 20 SQUARES X X mm3
5 20 10

= 1 mm3 N. CELLS
S

50
1 mm3 = 50 X 200 X N
= 10.000 X N
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/mm3
INFANTS 1-6 MOUNTHS 3.8 - 5.2 million/mm3
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)

- 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 5

2 4
DRO
1 P
3

SMEAR OF BLOOD

TAIL HEAD
(THIN) 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 2nd 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
 EVALUATION OF THE SMEAR
NOT GOOD

THE FILM IS SATISFACTORY BLOOD DROP IS SMALL

DRY IT BLOOD DROP IS TOO MUCH

GEIMSA STAIN
 FIXATION WITH 96% METHANOL 2 - 3’
 COVER THE SLIDE WITH DILUTED GIEMSA
SLIDE DO NOT FREE OF FAT
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
METHOD : - CENTRAL OF THE SMEAR = LYMPHOCYTE
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
- 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
- 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
- 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
- SHAPE : ROUND
- NUCLEUS : LARGE, OCCUPYING MOST
OF THE CELL, CHORMA-
TINE DENSE, DARK PURPLE

B. LARGE LYMPHOCYTE CELL

- SIZE : 10 - 15  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
- 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)

ALLOWED TO
BLOOD PLASMA
STAND FOR
+
ANTICOAGULANT ERITHROCYTE
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

METHODS : 1. WINTROBE METHOD

2. WESTERGREN METHOD

1. WINTROBE METHOD

STAND FOR
1 HOUR

BLOOD : Na.CIT.
( 4 : 1 )

MATERIALS :
- SYRINGE AND NEEDLE
- TUBE/ BOTTLE AND ANTICOAGULANT
SODIUM CITRATE
- PASTEUR PIPET
- THE WINTROBE TUBE

PASTEUR PIPET
 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
 SHAKE

1.6 mL BLOOD
0.4 ml 3.8% SOD. CITRATE

NOTES : THE HEIGHT OF THE COLUMN OF PLASMA IN mm

GRADUATION STARTING FROM THE 0 MARK AT
THE TOP OF THE TUBE

NORMAL : MEN : 0 - 15 mm / HOURS

WOMEN : 0 - 20 mm / HOURS
CHILDREN : 0 - 10 mm / HOURS

PATHOLOGIC : - ACTIVE TBC

- RHEUMATIC FEVER
- MALIGNANT DESEASE
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 1 1
X X mm3
20 20 10
1
WHEN
50 mm = N. CELLS
3

THE COUNT OF PLATELETS IN 1 mm3 = N X 50 X 100

= N X 5000 CELLS
 6B. INDIRECT METHOD (FONIO)

I.

GIEMSA STAIN

MATERIALS :
1. 14 % SOLUT. MgSO4 COUNT : ERITHROCYTE 1000 CELLS
THROMBOCYT : N. CELLS
2. BLOOD LANCET
3. SLIDES
4. GIEMSA SOLUTION

METHOD :
1. DROP SOLUTION 14 % MgSO4 ON THE FINGER TIP
2. PRICK AND MIX WITH BLOOD
3. MAKE BLOOD SMEAR AND STAIN BY GIEMSA STAIN
II. MAKE THE ERITHROCYTE COUNT
RESULT = Y. CELLS / mm3

N x Y
THE COUNT OF THROMBOCYT =
1000
/mm3

NORMAL RANGE = 150.000 - 400.000 / mm3

INCREASE = THROMBOCYTOSIS : - POLYCYTHEMIAVERA

- IDIOPATHIC THROMBOCYTHEMIA
- CHRONIC MYELOGENIUS LEUKEMIA
- FOLLOWING SPLENECTOMY

DECREASE = THROMBOCYTOPENIA : -THROMBOCYTOPENIA PURPURA

- APLASTIC ANEMIA
- ACUTE LEUKEMIA
- GAUCHER’S 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

COTTON

MIX
STAND FOR 15’
BLOOD 3 DROPS AND 370 C
OR EDTA VENOUS BCB SOL. 3 DROPS
BLOOD
MAKE PREPARATION

MOIST DRY

EXAMINE WITH MICROSCOPE

DIAFRAGMA IN EYE PIECE
FROM PAPER WITH APERTURE
ERY : PALE BLUE
COUNT :
ERYTHROCYTE = 1000 CELLS RET : SIZE > E
RET : N. CELLS GRANULES / RET
IT MEANS THE COUNT OF FINE/VIOLET
THROMBOCYT IS (N %O )

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 :

PLASMA
LIPID

15’ - 30‘ BUFFYCOAT

BLOOD EDTA
45 1 mmS 10.000 PLAT/mL
CENTRIFUGE
3000 RPM
ERYTHROCYTE

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%
 2. MICROHEMATOCRIT

SPECIMEN :
CAPILLARY BLOOD
WHOLE BLOOD WITH EDTA

MATERIAL :
- CAPILLARY HEMATOCRIT TUBES
- HEMOLANCET + COTTON 70% ETHANOL
- CREATOSALE (SOFT WAX)
- CENTRIFUS MiH
- READING DEVICE
METHOD :
1
2

1. PRICK THE TIP OF FINGER

2/3 - 3/4
WITH HEMOLANCET
2. FLOW THE BLOOD INTO
m THE CAPILLARY TUBE
1m FILL 3/4 OF TUBE
3  m
7c 3. PLUG THE END OF TUBE
WITH CREATOSEAL
4. CENTRIFUGE :
CREATOSEAL - 10.000 - 20.000 RPM
2 mm
- 5‘
READ : USING READING DEVICE
TOP OF PLASMA
TOP OF RBC

READING
DEVICE

THE BOTTOM OF RBC

NORMAL : MEN : 40 - 54%
WOMEN : 37 - 47%
P.C.V IS NEEDED TO ESTIMATE ABSOLUTE VALUE OF BLOOD :

 M.C.V. (MEAN CORPUSCULAR VOLUME)

PCV
= X 10 3 (76 - 96 3 )
ERI fl = FENTOLITER

 M.C.H.C. ( MEAN CORPUSCULAR HB CONCENTRATION)

Hb
= X 100% (32 - 36% )
PCV PERCENT

 M.C.H. ( MEAN CORPUSCULAR HB)

Hb
= X 10 g (27 - 32 g )
ERI pg = PIKOGRAM

THIN BLOOD FILM ARE USED :

1. DIFFERENTIAL COUNTING
2. STUDIES OF MORPHOLOGIC BLOOD CELLS
3. IDENTIFYING PARASITES
 EVALUATION OF MORPHOLOGIC BLOOD CELLS

ERYTHROCYTE
- NORMAL : SIZE UNIFORM 2. ABNORMALITIES OF SHAPE
 6 - 9  m (7  m) (POIKILOCYTOSIS)
SHAPE ROUND BICONCAVE
A. TARGET CELLS

FROM ABOVE FROM SIDE

B. CRENATED CELLS
NORMOCHROM NORMOCYTIC

1. ABNORMALITIES OF SIZE : C. SPHEROCYT

ANISOCYTOSIS
A. MICROCYTIC
4-6 m
D. ACANTHOCYT

B. MACROCYTIC E. BURR CELL

10 - 12  m

D. MEGALOCYTIC F. CRESCENT BODY

( 12- 25  m)

C. SHCIZOCYT G. OVALOCYT /
(FRAGMENT) ELLIPHTOCYT

H. SICKLE CELL

I. TEAR DROP APP.

J. LEPTOCYT

K. BIZZARE
3. ABNORMALITY OF COLOUR :

A. HYPOCHROM

B. HYPERCHROM

C. POLYCHROMATION

D. STOMATOCYTE

4. INCLUSION BODY

A. BASOPHYLIC STIPLING

B. MALARIAL STIPLING

C. SIDEROSIT

D. CABOT’S 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. DOHLE’S (INCL.) BODIES

INFECTION POSITION BURN

7. AUER RODS

8. BARR BODY

9. PELGER-HUET ANOMALY
( < 2 LOBES)
ARTEFACT :

CRENATED ROULEAUX SMUDGE

CELL FORMATION CELL
 BLOOD PARASITES

MALARIAL PARASITE

THIN BLOOD FILM THICK BLOOD FILM

DRY DRY

FIXATION WITH DROP WITH WATER : HAEMOLYSIS

96% ETHANOL

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
1/2 (S + D)
10’ . ON THE FORHAND ABOUT4 cm BELOW
THE ELBOW, MAKE A CIRCLE WITH DIA-
4 cm METER 5 cm.
EXAMINE AND COUNT THE PETERCHIAE
ESPECIALLY IN THE CIRCLE
RL + = IF THE CIRCLE CONTAIN MORE
THAN 10 PETERCHIAE

BLEEDING TIME TEST

A. DUKE METHOD
B. IVY METHOD
40 mmHg

EVERY
30 SECOND
x

2 1

MATERIAL :
LOCALIZATION :
1. HEMOLANCET AND
- FOREHAND
EVERY
COTTON + ETHANOL 70%
- PLACE BLOOD PRESSURE CUFF
30 SECOND 2. STOP WATCH
PRESS UP TO 40 mm Hg
3. FILTER PAPER
METHOD : - MAKE 3 mm DEEP SKIN PUNCTURE
1. PUNCTURE THE EAR LOBE DEEPLY START :
2. START STOP WATCH - BLOT THE BLOOD FROM PUNC-
3. EVERY 30 SECOND, COLLECT THE DROP TURE SITE ON A PIECE OF CIR-
OF BLOOD ALONG THE STRIP OF CULAR FILTER PAPER EVERY 30
BLOTTING PAPER SECOND WHEN BLEEDING CASES,
4. WHEN NO MORE BLOOD APPEAR, STOP THE WATCH AND RELEASE
STOP THE STOP WATCH THE BLOOD PRESSURE CUFF
5. RESULT CAN MADE : - RECORD THE BLEEDING TIME
A. THE TIME OF WATCH
B. COUNT THE NUMBER OFF DROP &
MULTIPLY BY 30 NORMAL : 1 - 7 MINUTE
NORMAL : 1 - 3 MINUTE
 COAGULATION TIME OF WHOLE BLOOD
(LEE & WHITE)

3 ml START THE STOPWATCH

AS SOON AS THE BLOOD
ENTERS - AFTER 3‘, REMOVE THE 1st TUBE
THE SYRINGE
 : 7 - 8 mm 1 ml AND TILT FOR 450
1 ml
- REPEAT EVERY 30 SECOND
UNTIL THE TUBE CAN BE
GLASS TEST
COMPLETELY INVERTED
TUBE
- C.T. DETERMINED BY CLOTTING
TIME OF TUBE NO. 2

NORMAL : 5 - 11 MINUTE
1 2

WATER BATCH 370 C

CLOT RETRACTION

 5 ml TEST :
- THROMBOCYTE FUNCTION NUMBER
- FIBRINOGEN CONCENTRATION
- PACKED CELL VOLUME
CLOT

C.R. = % ACE SERUM

FROM BLOOD VOLUME
1 HOURS
IF THIS SERUM = 3 ml
BLOOD = 5 ml

WATER BATCH 370

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
 BLOOD GROUPING

SLIDE TEST :

ANTI A ANTI B ANTI A+B

MIX & SHAKE

AGGLUTINATION : BLOOD GROUP

+ - + A
- + + B
+ + + AB

- - - O

TILE TEST :

+ + - ERY A

+ - + ERY B

- + + ERY X

SERUM X ANTI A ANTI B

 + + + 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 2SO4

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
 1. FORM
2. COLOUR
3. SMELL
4. CONSISTENCY
MACROSCOPIC 5. MUCUS
6. BLOOD
7. PUS
8. REST. OF FOOD
9. PARASITE

- CARBOHYDRATE
- PROTEIN
1. REST OF FOOD - FAT
2. ERITHROCYTE/ - VEGETABLES
EXAMINATION MICROSCOPIC LEUKOCYTE
OF FAECES 3. AMOEBAE
4. THE EGGS OF WORMS

1. BLOOD
CHEMIST 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)
- + (CO2) : FERMENTATION OF CARBOHYDRATE

5. MUCUS
 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

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)

PUT THE COVER GLASS ON IT

MIX

EXAMINE BY MICROSCOPE
 MICROSCOPIC EXAMINATION OF FAECES

1. REST OF FOOD
REAGENS CARBOHYDRATE

A. CARBOHYDRATE : LUGOL
AMYLUM / CH + IODIUM BLUE

DEXTRIN

B. 30 % ACETIC ACID RED MUSCLE

VEGETABLE

CONECTIVE
TISSUE

C. FAT SUDAN III

RED BALL STRUCTURE

2. AMOEBA + EOSIN AS NEGATIVE STAINING V

(BACKGROUND : RED COLOUR) K

AMOEBA COLOURLESS
E L
ERY / LEUKO

3. ERYTHROCYTE / LEUKOCYTE : EOSIN (1 - 2%)

MORPHOLOGY APPEAREANCE
SAME LIKE IN THE SEDIMENT OF URINE
 4. THE EGGS OF WORM

A. DRY PREPARATION
STREAK THE FAECES

MATERIALS :
- OBJECT GLASS
- PARAFIN LIQUIDIUM
- WOOD STICK

ALLOW TO DRY ALL

OF THE FAECES

COVER WITH MICROSCOPIC

PARAFIN LIQUIDIUM EXAMINATION
THROUGH OUT THE FAECES

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 ANKYLOSTOMA
LUMBROCOIDES TRICHOCEPHALUS 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

METHOD :
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

MICROSCOPE
 C. CHEMICAL EXAMINATION

1. OCCULT BLOOD EXAMINATION

SMALLER INCREASES IN BLOOD CONTENT MAY NOT ALTER
APPEAREANCE OF THE STOOL
1.A. BENZIDINE TEST
REAGENTS :

1. BENZIDINE
2. ACETIC ACID GLASIALE
3. H2O2 3%

METHOD :
FILTER

FILTRAT
FAECES SUSPENSION
ml
2
1 gr
BENZIDIN

3 ml

ACETIC ACID 3 ml

+ : GREEN TO BLUE
READ WITHIN 5 ‘

H2O2 3%

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.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% H2O2

PROCEDURES :

MIX MIX
MIX WELL WELL
WELL 2 ml H2O2 3%

0.5ml 2 ml GUAIAC
2 ml WATER Gl.A A SOLUTION
0.5 gr FAECES

MIX
OBSERVE FOR 2 MINUTE
STARTING
RECORD THE MAXIMAL BLUE COLOUR
TIMER

TRACE 1 + 2 ++ 3 +++ 4 ++++

 2. STERCOBILIN

UROBILIN
PALE RED

STAND FOR
SEVERAL HOURS
(ONE NIGHT)
5 ml HCl
CONCENTRATE
BOIL
READ SOON
FAECES 1 - 3 gr BILIRUBIN
BLUE COLOUR
 SPUTUM

= 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

BE CAREFUL, BECAUSE THIS SPECIMENT (SPUTUM) IS INFECTIOUS

THE METHOD OF HANDLING AND DISCHARGE MUST BE STERILIZE
OR BURNED

EQUIPMENT :

- WORKING TABLES SHOULD HAVE BEEN CLEANED

- MICROSCOPE WITH DISINFECTANT LIKE
- AND OTHERS 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

PORTION
SECOND LESS VISCID FOAMIC LAYER

FIRST VISCID CLOUDLY FLUID LAYER

THIRD MOST VISCID 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

CEREOUS PLASMA
BODY CAVITY
FLUID 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 :

TRANSUDATE (INFLAMATION NEG. -)

UNKNOWN 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 : RIVALTA TEST

FLUID
(EXUDATE / TRANSUDATE)

RESULT :
1 cm
- = CLEAR (NOT TURBID)

RECORD THE REACTION

TURBIDITY OF SOLUTION
+
(WEAK)
= RATHER TURBID (FINE)

AQUADEST
+
+
(STRONG)
= REAL TURBID, PRECIPITATE
DENSE FOG,
1 GTT GLACIALE ACETIC ACID

- 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 SS S

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
 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
DANGER ! :
- INTRACRANIAL PRESSURE
- INFECTION
LUMBAL PUNCTURE - PAIN
L3/L4

STERIL
TUBE

TO PREVENT
SPONTANT
COAGULANT

DISCARDED EXAMINATION BACTERIOLOGY

20% Na Citrat
0.01 ml/ml LC
MICROSCOPIC IMMUNOLOGY/
MACROSCOPIC SEROLOGY
CHEMISTRY
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 :
FROIN’S SYNDROME : - XANTHOCHROM
- PROTEIN HIGH
- PLEIOSITOSIS LYMPHOCYTE

5. PRESSURE : N = 70 - 200 mm H2O

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 mm2
DEPTH = 0.2 mm
2. CONCENTRATE TURK SOLUTION

METHODS : - DRAW TURK = 1.0 9

DILUTION
- DRAW CSF = 11.0 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
N 10 10 50 N N
LEUKOCYTE COUNT : X X = =
16 2 9 144 3
NORMAL : 0 - 5 / mm3 CONSIST OF LYMPHOCYTES
( < 5TH : 0 - 20 / mm3)
ABNORMAL : > 10 / mm3
M. PURULENTA : 500 - 20.000 mm3
M. SEROSA : SPI 200 / mm3

WHEN USED AN IMPROVED NAUBAUER

CALCULATION : 1 9
VOL = 9 X = mm3 = N. CELLS
10 10
10 10 100 5
CELL COUNT : N X X = = N
9 9 81 4

ERYTHROCYTE : NORMALLY ABSCENT

 THE OTHER COUNTING METHOD :
1. MIX CSF = 0.05 ml
DILUTION = 1/20
TURK = 0.95 ml
2. COUNT THE CELLS IN 5 mm2 (USING SQUARE 1,4,7,13,16)
1,4,7,13,16
WHEN = N. CELL
TOTAL CELLS = 20 X N

FUCH ROSENTHAL HEMOCYTOMETER

 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

NONNE’S TEST
FOAM TEST PANDY’S TEST (NONNE APELT)
ROSS JONES

1 DROP CFS

CFS aa
REAGENT :
CFS 1 ml REAGENT

1/2 - 1 ml REAGENT
R/
R/ AMMON. SULFAT
PHENOL LIQUEFACTUM 10ml
SHAKE AQUADEST 90 ml
CONCENTRATE IN WATER
HARDLY
RESULT :
N : 1 - 2’ FOAM + : WHITE CLOUD STAND FOR 3’
DISAPPEARED
AN : 5’ REMAIN - : NO WHITE CLOUD OR
SLIGHT CLOUDNESS LOOK RING

+ : APPEAR WITHIN 3’
MEAN : SHAKE, DISAPPEAR
HIGH PROTEIN CONTENT
GLOBULIN
++ : SHAKE, OPALESCENT
+++ : SHAKE, CLOUDLY
++++ : SHAKE, THICK CLOUDY

ALBUMIN
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
GLUCOSE DECREASEED
M. TBC

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

SMEAR PREPARATION
DISCHARGE OF (PUS) :
GRAM STAINING
 URETHRA
- GO CULTURE
 VAGINA
 CERVIX - CANDIDA ALBICANS KOH
- TRICHOMONAS VAGINALIS

1 GTT SPECIMENT + 95% SALINE

MICROSCOPE
DETECT FLAGELLA
(MOTILITY)

 PROSTATIC SECRET COLLECTED BY MASSAGE

 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:
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 mm2) = N. CELLS
- RESULT = SPERM / ml = N X 50.000

NORMAL : 70.000.000 - 300.000.000 / ml

2. MOTILITY OF SPERM

OBSERVE 200 SPERM

- SCAN SEVERAL FIELD 40X SMALL DROP
TOTAL + 200 SPERM
- FOCUS THE ENTIRE DEPTH
NON MOTILE SPERM CEMEN
370 C
- RECORD % OF MOTILITY SPERM

N : 70 - 80 % ACTIVE ( WITHIN 1 HOUR)

 3. MORPHOLOGY

- PREPARE THE SMEAR

- STAIN WITH : GIEMSA, FUCHSIN. ETC
- + 200 SP EXAMINE 200X

TAIL HEAD
3-6m
50 - 70  m
NECK
ABNORMAL :

IMMATURE SPERMATOZON
(SPERMATID)

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

CLINICAL PATHOLOGY
ROUTINE & SIMPLE
DEPARTMENT OF :
CLINICAL LABORATORY
1. CLINICAL HAEMATOLOGY
- URINALYSIS 2. CLINICAL BIOCHEMISTRY
- HAEMATOLOGY 3. CLINICAL “ROUTINE” EXAM
- FAECES 4. CLINICAL MICROBIOLOGY
- CSF/BODY FLUIDS 5. CLINICAL IMMUNOLOGY
6. BLOOD BANK
7. EMERGENCY

PHYSICOLOGY

MANAGEMENT
BIOLOGY CLIN. PATH LAB.
SUCCES PERFORMANCE :
TO EFFECTION
REACH &
AMBISION EFFICIENT BASIC
AND MEDICAL
TARGET SCIENCE

MEDICAL
PRACTISE
 DESIGN OF MANAGEMENT
FROM ROBBIN
CONFLICT
POLITICS POWERS RESOLUTION

ORGANIZING
SATISFACTORY
O
I FINANCIAL PLANNING PERFORMANCE
U
N
T
P PHYSICAL DIRECTING PRODUCT
P
U
U
T HUMAN CONTROLLING SELF SERVING
T
BEHAVIORS
STAFFING

CONFLICT
POLITICS POWERS 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
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

BRIDGING

MEDICAL PRACTICE
BASIC SCIENCE DIAGNOSIS
RESEARCH THERAPY
EDUCATION PROGNOSIS
QUALITY ASSURANCE

REAGENTS
EQUIPMENTS
COMPUTER SCIENCE
MANAGEMENT TECHNIQUES
INDUSTRY