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2546

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5
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25




1.
2.

3.

3.1 (infectious agent)
3.2

3.3
centrifuge
4.
4.1

4.2
4.3
4.4

4.5

4.6

4.7

4.8
4.9 /
4
7773 ()

1.
1.1
1.2


1.3

1.4
2.
2.1
2.2 3
2.3 dropper 1
3.
3.1
3.2


1.
1.1

1.2
(absolute alcohol ether 3 7)

1.3
2. centrifuge
2.1 95% alcohol

2.2
3. Hematocrit
3.1 hematocrit tube
3.2 95% alcohol
3.3
4. Hematocrit
4.1 95 % alcohol
4.2
5. Refractometer
5.1 cover plate prism

5.2

1. (infectious waste)




2.



3.
3.1
3.2
3.3

- Gram stain:

1.
2.
3.
4.

centrifuge
Hematocrit
Hematocrit
5. Refractometer

1.
2.
3.
4.
5.
6.
7.
8.
9.
10.
11.

()
()

(Virkon 1:1000)

12.
13.
14.

15.
16.
17.
18.
19.
20.
21.
22.
23.
24.
25.
26.
27.
28.
29.

Clamp
Hematocrit capillary tube
Medicine dropper
Pasteurs pipette
Rack

Gentian violet
Gram iodine
95 % alcohol
1% safranin
- Wright stain:
Wright stain reagent
- Acid fast
Kinyoun reagent Acid alcohol
modified stain:
Methylene blue 2% sulfuric acid
- India ink stain:
India ink
- White cell count: 10% acetic acid
- Urine examination: Roberts reagent Benedict reagent
- Others:
Normal saline
Oil immersion
Pandy solution Sterile water

Blood
- Septicemia

- FUO
- Subacute
endocarditis

Aseptic technique
2 .
- hemoculture 2-3
5 15 - 1 .

- hemoculture 3
5 24 .
1-2 .

Body fluid
- Aseptic technique 15

*
14.00 . 48o C

Respiratory tract
- Nasopharynx,
Pharynx

- swab NSS
transport
media
- Bronchoalveolar -
lavage
- Sputum
- 2-3

- Endotracheal
- suction NSS 2-3
aspirate
suction
Genital tract
- swab NSS
transport
media

2 .

- Stool

- Clean voided
midstream
- Indwelling
catheter
- Straight
catheter

2 .

- swab anal sphincter

2 .
2.5 . transport media
-

-


- aseptic technique syringe
catheter 5
.
- aseptic technique
15 .

- swab NSS

transport media
-
aseptic technique
- anaerobe
()

- aseptic technique
IV catheter sterile
NSS

( formalin )

2 .
4oC

Urine

Pus

Gastrointestinal tract
- Rectal swab

-
-
IV catheter

2 .

15


Hematocrit
Plasma
Buffy Coat = White blood cells
Red blood cells

> SCALE

100
90
80
70
60
50
40
30
20
10
0

1. alcohol

capillary tube
3 4 tube
2. EDTA
capillary tube capillary
tube 3 4 tube
3.
4. hematocrit 10,000 13,000
5 ( Balance tube )
5. hematocrit
6. capillary tube

White Blood Cell (WBC) Count

1

4
1 .

1 .
WBC count chamber
4 chamber
2

1. EDTA
2. WBC count pipette 0.5
(10% acetic acid) 11
3. pipette 3
4. pipette 2-3 chamber
5. 10
diaphragm
6.

7. chamber

1 = 1 x 1 x 0.1 .. = 0.1 ..
4 = 0.1 x 4 ..
N = N / (4 x 0.1) ..
dilution = 20
= (20 x N) / (4 x 0.1) = N x 50 ..
WBC = 4,50011,000 .. (4.511 x 109 / L)

nucleated red cell (NRC) blood smear

leukocyte (WBC) count

 Blood Smear

Neutrophil

Eosinophil

Large lymphocyte Small lymphocyte

Basophil

1.

2. blood smear
3. Wright blood smear
4. 4
5.
6. 4
7. blood smear
8. 10
100
9.

Monocyte

10

11

12

13

14

15

16

17

18

19

20

21

22

23

24

25

26

27

28

29

30

Plasmodium falciparum 1: Normal red cell; 2-18: Trophozoites (2-10 are

ring-stage trophozoites); 19-26: Schizonts (26 is a ruptured schizont);
27, 28: Mature macrogametocytes (female); 29, 30: Mature
microgametocytes (male)

Thin film Fields stain

1.
2.
3.
4.
5.
6.
7.

blood smear
fix methanol 3
Fields stain solution B 1-5

Fields stain solution A 1-5

Thick film Wrights stain

1. 2-3
2. smear ~ 2 .

3.
hemoglobin
4. Wrights stain 1 2
3
5.
6.

10

11

12

13

14

15

16

17

18

19

20

21

22

23

24

25

26

27

28

29

30

Plasmodium vivax 1: Normal red cell; 2-6: Young trophozoites (ring

stage parasites); 7-18: Trophozoites; 19-27: Schizonts;
28, 29: Macrogametocytes (female); 30: Microgametocyte (male)

1. Thin blood film

-
200-250 oil fields
- oil immersion 100
oil fields 8-10
- species

- 1,000 RBC species
2. Thick blood film
- WBC 3-5 oil field

- oil immersion 50 oil

fields 10
-

- 200 WBC species

plus system
1 +
1-10 100 oil fields
2 +
11-100 100 oil fields
3 +
1-10 oil field
4 +
> 10 oil field

7.

(Urine specific gravity)

1.355

1.050

1.350

1.040
1.030

1.345

1.020

1.340

1.010
1.335
1.333

1.000
U.G.
ml

20 C

ND

refractometer
urine specific gravity
1. NaCl
2. Urea
3. Creatinine
4. Glucose
5. Radiocontrast agent (Omnipague)
6. Albumin

Molecular Weight (Da)

58.5
60.0
113.12
180.0
821.14
68,000.

1. cover plate prism refractometer

1
2. cover plate 1
prism cover plate eyepiece
1.000
3. cover plate prism
4. cover plate 1 prism cover plate
5.
6. cover plate prism

1. Urine specific gravity (density)

solute
(molecular weight) urine osmolality particle
number
2. urine specific gravity

10

Urine Protein

neg

trace

False positive

1+

2+

3+

4+

False negative

Contrast media
Highly buffered alkaline urine
Antibiotics in high concentration,
Dilute urine
e.g. penicillin and cephalosporin
derivatives
Uncentrifuged turbid urines false positive false
negative - urine supernatant

1. 2-3 .
2 45 o pipette Roberts
reagent
Roberts reagent
3
4

Negative
Trace
1+
2+
3+
4+

11

Urine Sugar
Benedict 5 .
8 0.5 .

alcohol 5

neg

False positive

trace

1+

2+

3+

4+

False negative

Presence of hydrogen peroxide,

Ascorbic acid
bacterial peroxidases (e.g. cystitis), salicylates
hypochlorite chlorine
tetracycline
false positive reactions
Formaldehyde

Negative
Trace
1+
2+
3+

4+

12

Urine Sediment

dysmorphic RBC

heme cast

oval fat body

waxy cast

RBC cast

hyaline cast

granular cast

fatty cast

dirty brown cast

broad cast

1. 10 .
2. centrifuge 2,000 5
3. 1 .

4. pipette 1
cover slip
5. 10
40
6.

7.

1. Nephritic urine: proteinuria

dysmorphic RBC, RBC cast heme cast
2. Nephrotic urine: proteinuria , oval fat body, fatty cast, etc.
3. Nephrotic-nephritic urine: 1 + 2
4. Telescopic urine: 1 + 2 broad cast
5. Acute renal failure: tubular cell dirty brown (muddy) cast
6. Chronic renal failure: proteinuria , broad cast

13


Direct Smear
Ascaris

Hookworm

Trichuris

Strongyloides larvae

H. nana

H. diminuta

Opisthorchis

Taenia

Capillaria

Fasciolopsis buski

Giardia lamblia

E. histolytica cyst

1. (NSS) 1
2.

3. smear
4. cover glass cover
glass

5.

8. 1
% cover glass
9.

10.

high power field

14


dilute (1:10)

11

10% acetic acid

11

11

0.5

0.5

1.
1.1 WBC chamber 9
1.2 .. 10 9
2.
2.1 WBC count pipette 0.5 1

2.2 10 % acetic acid 11 (dilution 1:20 1:10
)
2.3 3-4 WBC chamber
2.4 .. dilution factor
3.
3.1 WBC RBC

3.2 Dilute 10 % acetic acid (dilution 1:10) WBC 9

.. 10
3.4 RBC

15

Differential Cell Count

Neutrophil

Lymphocyte

1 pipette
2 500-1,000 3
3. ( protein sugar
)
4.
5. pipette 1 smear
6.
7. Wright stain
1:1
8. 3
9.
10.

Eosinophil
16

India Ink Preparation

Cryptococcus
1. pipette
2. 1,000 3
3. ( protein
sugar )
4. pipette 1
5. India ink
6. cover glass
7.

- background yeast cell

background capsule
- yeast cell budding
Cryptococcus

India ink preparation. 1: yeasts, 2: budding yeast, 3:

white blood cell, 4: artifacts
17

Wrights Stain Effusion

Staining of Effusion. 1: eosinophilic pleuritis, 2: mesothelial cell,

3: mesothelioma, 4: adenocarcinoma, 5: metastatic carcinoma,
6: lymphoblasts small lymphocytes

effusion (~ 20 . effusion
~ 40 50 . effusion ) 700 1,000
5
2. Wintrobe tube
3.
4. 1

5. Wright
5.1 Wright (5 8 )
5.2 2 3 5
smear WBC 2
5.3
5.4
6.

7. effusion tube

18

semi-fluid sample

fluid sample

fix

1. smear
smear 1 .
- CSF, fluid
3,000 10-30 sediment
smear
- dropper 1 smear
- tissue
2.
3. Fix 3-4

1.
smear
2. smear


3. film smear fix

19

 Grams Stain

Gram +ve cocci

in cluster

Gram +ve cocci

in chain

Gram -ve diplococci

Gram -ve bacilli

1. gentian violet smear 1530

2.
3. Gram iodine 15-30
4.
5. 45 95 % alcohol
smear
6.
7. 1% safranin 10-15
8.

1. : epithelial cell, white blood cell, bacteria,

fungus
2. Gram : nucleus cytoplasm
smear
3. bacteria
Gram positive
Gram negative

20

 Acid Fast (Kinyoun method)

1.
2.
3.
4.
5.
6.

Mycobacterium tuberculosis

Kinyoun smear 5

(decolorize) acid alcohol

methylene blue 1 2

1. bacteria normal flora

acid fast bacilli
2. 300 oil
fields (F) WHO/UNAIDS

0
1-9 100 F
10-99 100 F
1-10 F ( 50 F)
> 10 F

AFB: not found

AFB: bacilli
AFB: 1+
AFB: 2+
AFB: 3+

21

 Modified Acid Fast

Nocardia

Actinomyces

1.
2.
3.
4.
5.

smear
Kinyoun 5
( )
2 % sulfuric acid
2 % sulfuric acid 3

6.
7. methylene blue 1-2
8.

1. Nocardia

branching filament
fungal hyphae
2. Actinomyces branching filament
3. Rhodococcus cocci coccobacilli

Rhodococcus
22


Potassium Hydroxide (KOH) Preparation

Budding yeast

Malassezia furfur

, nail bed

1. 10% KOH 1
2. 10 % KOH
3. cover glass
4. (, KOH
keratin) 10 20
5. KOH ()

1. objective lens 10 40
diaphragm epithelial
cell
2. ( Malassezia furfur) budding yeast
hyphae (meatballs and spaghetti appearance)
3. ( dermatophytes) branching septate hyphae
/ arthroconidia

Dermatophyte
23

Tzanck Test

Tzanck preparation
vesicular lesion multinucleated giant cell

Wright
1. vesicular lesion smear
( vesicle
vesicle )
2.
3. Fix 95 % alcohol 1
4.
5. blood smear

- epithelial cells multinucleated cell

- polymorphonuclear cells, lymphocytes,
mononuclear cells
- smear intranuclear inclusion bearing cells

Giemsa

- fix absolute methanol 5-10

- Coplin jar diluted Giemsa
stain ( 1:20-1:40) 95 % ethanol

24

Sleep Lab

1
2
2
2
5
2
2
2
1

4
4

72 4
6

7745-6
7755
7758
7760
7791
7295
7293
7293
7281-3
119
8382
7203,
7301
7100
7642-4
15, 16

.. ()
.. ( )
..
..
..



..

, .. ,

..

25