,

11000
0 2951 000 99305 0 2591 5449
www.dmsc.moph.go.th
1 2556 500
2 2557 30,000



. :
, 2557. 56 .
1. . I. , . II. .
681.761
ISBN 978-616-11-2130-3

II

(
)
( )

2556
2556

III

(Biological safety cabinets)

(Biological safety cabinets)

(US CDC Atlanta)

-

CDC
Atlanta)

(US

2556

2556

IV

3 ( 3 )

3
(
3
)

()

2556
2556

VI

III

IV
V

?
?

3 1
3
13
12
16
14

25

26

30

32 28


3631

33
34
40 39
46 40
48 48

62

VII

VIII

1.

2. Laminar flow clean bench


3. 100%


4.

5.


(Biosafety
Cabinet Technology) ?

Biological Safety Cabinet Biosafety Cabinet
BSC BSC

-
Stainless 2



-

- HEPA filer

- (Laminar airflow)
(Blower) HEPA filter

(Good Microbiological Technique) BSC
(Standard Microbiological
Practice)

 BSC 3

HEPAfilter BSC
(
)

3
Blower/fan/motor
))

HEPA filter((

Blower/fan/motor ()
HEPA filter
HEPA filter
Laminar flow

Laminar flow
HEPA filter
Laminar flow

Blower
1 (BSC) 3
(: WHO laboratory biosafety manual 2004)

3
3

Laminar ow
lter
HEPA filter
HEPA
Blower

Laminar flow
Blower

HEPA filter
1. HEPA 1.filter

HEPA filter ?

HEPA filter HEPA

filter High Efficiency Particulate Air filter HEPA

HEPAfilterfilter
Efficiency
Air fibfilter

HEPAfilter

High
Boron
silicate Particulate
fiber Glass
er

Boron silicate
Glass
fiber

Fiber
fiber
Fiber

(Air space)

(Air

Fiber
0.3 Filter

Filter
Fiberspace)

0.3

(
2)

(Open
space)

( 2)

(Open
space)
(Pore
(Poresize)size)

(Pleating)
(Pleating)

Aluminium
Aluminium( 3)

BSC
( 3)
BSC

(Frame)
(Frame)

(Leak)

filter
BSC
(Leak)

HEPA

HEPA filter BSC

(: URL:bestvac.net/howto)

(: URL: biologicalcontrols.com/800400.shtml)

2 HEPA filter HEPA filter 500

/
Continuous
sheet of filter
medium

Filter frame

Continuous
sheet of filter
medium

Filter frame

Aluminium
Seperator

Aluminium

Seperator
3
HEPA
filter
3 HEPA filter
(: URL:air-purifier-reviewsite.com/featured/air-purifier-technologies-hepa-filter)
(: URL:air-purifier-reviewsite.com/featured/air-purifier-technologies-hepa-filter)

HEPA filter

! !
HEPA filter

HEPA fiHEPA
lter filter
(Air(Airfilter)filter)

HEPA
fi
l
ter

(Contamination)
HEPA filter
(Contamination)

 HEPA filter !

HEPA filter (Air filter)
HEPA filter
(Contamination)


HEPA filter BSC

(Pore size) HEPA

(Micron)

50-150 0.01-100
0.001-10 0.0001-0.01 HEPA
0.3

HEPA filter !

HEPA filter Pore size 0.3
... HEPA filter
(Sieves)
HEPA (Pore size)

Pore size
HEPA filter
!!

Pore size 0.3

HEPA filter (Sieves)

HEPA

(Pore

size)

HEPA

HEPA

4


4
1. Sieve effect Pore size Filter

(0.3

(0.3

1. Sieve effect

)
Pore size
Filter
)

Sieve effect
Sieve effect
Sieving

Sieving

Particle

Media Fiber
Flow
Media Fiber

Particle

Media Fiber

4 Sieve effect
(: URL:Air-purifier-reviewsite.com/featured/air-purifier-technologies-hepa-filter/)

2. Inertial impaction 0.3

2.Inertial

2. Inertial
impaction

0.30.3

impaction

HEPA

Filter

impaction

Filter
0.3

Filter 2. Inertial

...
Inertial
impaction
Inertial
impaction


Inertial impaction
Filter
Flow
Particle
Particle
Media
Fiber
Media
Fiber
Inertial impaction

Flow 7

5 Inertial
impaction impaction
5 Inertial
Flow
(: URL:Air-purifier-reviewsite.com/featured/air-purifier-technologies-hepa-filter/)
Particle
Media Fiber
(3.: Interception
URL:Air-purifier-reviewsite.com/featured/air-purifier-technologies-hepa-filter/)

impaction
0.1
5 Inertial
(
: URL:Air-purifier-reviewsite.com/featured/air-purifier-technologies-hepa-filter/)

3. Interception

0.1

3. Interception

0.1

3. Interception

0.1
Filter

FilterParticle

Interception
FilterFilter

Particle
Flow
Particle
Interception Interception
Interception
Flow
6 Interception
Flow
(: URL:Air-purifier-reviewsite.com/featured/air-purifier-technologies-hepa-filter)

4.

4. Brownian diffusion 6 Interception

0.1
(
: URL:Air-purifier-reviewsite.com/featured/air-purifier-technologies-hepa-filter)

4. Brownian
diffusion

0.1
Zigzag Fiber

6 Interception

Brownian
(
: URL:Air-purifier-reviewsite.com/featured/air-purifier-technologies-hepa-filter)
Zigzag Fiber
Brownian diffusion
0.1
Brownian
7

7 Diffusion
Zigzag(:

Fiber

URL:Air-purifier-reviewsite.com/featured/air-purifier-technologies-hepa-filter)

Flow
6 Interception
(

:
URL:Air-purifier-reviewsite.com/featured/air-purifier-technologies-hepa-filter)
4. Brownian diffusion 0.1

Brownian

Zigzag

0.1


4.
diffusion

Fiber

Zigzag Fiber
Brownian
Brownian

7 Diffusion
(: URL:Air-purifier-reviewsite.com/featured/air-purifier-technologies-hepa-filter)

Pore size HEPA filter

HEPA filter

Pore size HEPA filter

HEPA filter

8 HEPA filter ?

HEPA filter
Filter
HEPA filter Filter

HEPA 99.97% 0.3

0.3 99.97%
0.03% 0.3 (
) ( 8)
0.3 99.97% 0.3
100%

HEPA

4 ...



0.3
(

(8 )

0.3
99.97%

8 )

0.3
99.97%
(0.3

100%
0.3 100%
Overall Efficiency

Overall Efficiency

99.97
Diffusion
Diffusion
Interception
Interception
Impaction
Impaction

Particle
ParticleDiameter
Diameter(microns)
(microns)
HEPA

88

HEPA

Specification HEPA filter

- BSC (NSF 49) BSC class II HEPA filter type C

99.99% 0.3
- BSC (EN 12469) Filter type H14 99 99.995%
0.3

HEPA filter PAO
0.3 Filter filter
Filter

HEPA
filter

ULPA
filter

HEPA filter PAO

ULPA filter PAO

 HEPA filter !

ULPA filter 99.999-99.9999%
0.1-0.2 HEPA

ULPA filter Semi-conductor ULPA
HEPA filter BSC BSC

HEPA filter
ULPA ULPA filter

()

HEPA filter
Polystyrene latex ULPA filter
ULPA HEPA
2.
(Biological
safety cabinet blower)
Filter

HEPA filter BSC

safety
cabinet
(Blower)blower)
(Motor)
2. (Biological

blower)
BSC
2.

(Biological
safety
cabinet
Type

(Blower)

II
class II

(Blower)

(Motor)
(Motor)

class

A2,

BSC

Type
A1,
B1

Type
B2

BSC
classII class
II Type
class II


Type

9)A1,
II



(Type

B1 class
Type
B2
Type A2, B2

A1,TypeB1 A2,

( 9)

( 9)

Type
with canopy
canopy
Type A1
A1 Cabinet
Cabinet with

Type
Type B1B1

Type A1 Cabinet with canopy

Type B1

Exhaust

Exhaust

Exhaust

Inflow

Exhaust

Inflow

Inflow

Inflow

Blower
Blower

Blower
Blower

Type
withwithcanopy
canopy
TypeCabinet
A2 Cabinet
canopy
Type A2
A2
Cabinet
with

Type
Type B2
B2B2
Type

Exhaust

Exhaust

Supply

Supply

Blower

Blower

Blower

Laminar flow

Exhaust

Exhaust

Blower

Blower

Inflow

Inflow

Inflow

Inflow

9 Blower BSC A1 A2 B1 B2
(: Eagleson Institute)

10

9 Blower BSC A1 A2 B1 B2
Blower
(: Eagleson
Institute)

laminar ow

Blower

laminar ow

11

3. Laminar flow

3. Laminar flow

Laminar flow !
flow

LaminarLaminar
flow3. Laminar
flow

Laminar

Laminar
flow

flow

Laminar flow !

Laminar
BSC
flow
Laminar
flow

BSC

(Air
stream)

(Air
stream)

(flow
10)( 10)

Laminar

(Operator)

(Environ

BSC

(Operator)

ment)

(Air
(Product)

(Environment)
stream)

( 10)
(Product)
(Operator)
(Environment) (Product)

Laminar
flow
Laminar flow
10 Laminarflow
flow (
(:: Eagleson
10 Laminar
Eaglesoninstitute)
institute)

Laminar

Laminar
flow flow

BSC
BSC
-

-

Laminar

BSC
-

flow

10

10

10

3
BSC


! inar

Lamow 3 BSC !

3 BSC
Laminar ow
! inar

12

Lamow

BSC,
lab

Laminar ow

12

BSC,
Laminar

lab
Laminar flow
flow

BSC
lab

11

1. Biohazards

Hazards

Biological
( )
(

)


(Infectious agent) Bacteri
1. Biohazards
Parasites,
Fungi Rickettesia


BSC

Biological
( ) Hazards (

)
(Infectious agent) Bacte
Parasites, Fungi Rickettesia
1. Biohazards

11

Biological (2.)Aerosol
Hazards

()

(Infectious agent)
Aerosol
!


11

Bacteria, Virus, Parasites, Fungi Rickettesia

Aerosol
2.
Aerosol

Aerosol

2. Aerosol

AerosolAerosol

Aerosol

Aerosol
Aerosol !

Aerosol

Aerosol Aerosol

Aerosol
12 Aerosol
Aerosol Aerosol
Aerosol

3. Containment

12 Aerosol

(Method) (Facilities) (Equipment)
Containment

(Standard microbiological practice)


Biosafety level (BSC) : BSL1,
BSL2, BSL3, BSL4

BSC

12




BSC

4. Primary containment Secondary containment

Primary containment
BSC Secondary containment

5. Risk group ()

(WHO) 4 Risk group 1, 2, 3 4

Risk group 1
Bacillus subtilis, Infectious canine hepatitis E.coli

Risk group 2
Measles virus, Saimonellae
Hepatitis B virus

Risk group 3
M.tuberculosis, St.Louis Encephalitis, Eastern Equine encephalitis virus Chikungunya

Risk group 4
Marberg, Ebola


!

Standard practice,
BSC, BSL
Containment

13

 BSC ()
-

BSC
(Air tight)

BSC (
)
-

(Flush)

)(Laminar
flow)
-

BSC
(

(Air tight)

(Air
-

tight)

(Front

(Flush)

flow)

(Flush)

(Laminar
grille)

(Laminar flow)

(Rear


-

(Front

grille)
(Front grille)
grille)

(Rear grille)

grille)

(Rear

(Protect

(Protect

(Protect environment)
operator)
operator)
(Protect
environment)

(Protect
Product)
(Protect
(Protect
product)
operator)
(Protect environment)

Window)

product)


(Sash(Sash
Window)

(Protect

(Sash Window)
-

BSC

13 Grille

13 Grille

BSC (Class)

16 3 Class I, Class II Class III

16

14

Class

III
BSC

Class
4
Class
II II 4

Class

BSC Class I
: (Operator protection)
BSC
Class I protection) (Product)
(Environment

: (Operator protection) (Environment protection)

(Product)
Class

: Centrifuge
: (Aerosol)


Aerosol

Tissue
Centrifuge

homogenation
I

(Aerosol)

Class
(Hard-ducted)

Aerosol

Tissue

Class I

Class I homogenation)

(Hard-ducted)
Class I
Exhaust filter

17

Front
Opening

Plenum

Room air
Contaminate air
HEPA-liltered air
Side View
14 Class I
( : BMBL 5th edition, appendix A)

HEPA filter (Exhaust system)

Airflow
1. (Airflow) (Laminar flow)
Chemical fume hood HEPA filter
(Exhaust filter)
(Environment protection)
2. (Product
protection)

3. (Room air)

15

HEPA filter : (Exhaust system)

Airflow :

1. (Airflow) (Laminar flow)
Chemical fume hood HEPA filter (Exhaust filter)

(Environment protection)

2. (Product protection)
BSC3.
Class II
(Room air) (Inward airflow

Inflow)
(Front opening)

:

(Operator
protection)

(Operator protection) Inflow

(Environment protection)
(Product
protection)

BSC
Class
II
Class
II Class
II 4 (Type) Type A1,
BSC
BSC
:
II (Operator
(Operator
protection)
(Environment
protection)
:
Class

protection)

A2, B1
B2(Environment

protection)

protection)

(Product

:(Product

protection)

(Operator
protection)
4

Class
II




Class

II

(Type)

Type
(Environment protection) (Product protection) A1,

(Type) Type A1, A2, B1 B2

A2, B1

Class B2

II

4 (Type) Type A1,

A2,
B1

B2

A1
A2
B1
B2
A1 A1
A2
B1 B2B2
15
BSCA2 Class
II B1

15

BSCClass
Class IIII

15

BSC

: Class II

: :Class
Class
II

II

BSL3

: BSC
II

BSL1
BSL2
BSL1 BSL2 BSL3

BSL1 BSL2
BSL3

BSL2
BSL3

BSL4

( 15-16)BSL1

BSL4

Positive

BSL4
Positivepressure
pressure personnel
personnel
suit

BSL4 BSL4

BSL4
pressure
personnel
suit

BSL4

BSL4
Positive
suit

positive pressure
personnel
Class suit
II (
21) Class II

Class
II Class
(II 21)

( 21)

16
(Coveralls)

class II
BSL III
Operator
protection
16 Positive suit Class
II
( : En.wikipedia.org/wiki/
Product
protection
( : En.wikipedia.org/wiki/biosafety_level)
Envinronment protection
biosafety_level)
16 Positive suit Class
II class II
3 protection
( : En.wikipedia.org/wiki/biosafety_level)

3P:

3P:

Positive suit Class II

16 16
( : En.wikipedia.org/wiki/biosafety_level)

operator protection
Product protection
Envinronment protection
class II

operator protection
Product protection
Envinronment protection

19

 BSC

HEPA filter : Class II HEPA filter 2
(Exhaust filter) (Supply filter) Supply filter
1

Airflow : Class II Laminar Airflow ()
HEPA filter (Sterile cell culture) (Virus
propagation) (Product protection)

- Inward / Inflow airflow

(Room air) BSC
(Front grille)
(Front opening)
Inflow Type (A1, A2 B1 B2)
Room air Type B2 Room air

- Downward airflow

BSC HEPA (Supply filter)
Downward airflow

(Split) 2 2-6
(Rear grille)
Front grille
Exhaust filter Supply filter

Supply filter (Re-circulating air) 17-20

Inflow
Inflow

Down
flow
Down flow

17

Sash
Sash
Front grille
Inflow
Front grille
Inflow
Sash

Exhaust
Exhaust
Supply Filter
Supply Filter
Downflow
Downflow
Exhaust
Supply
Blower Filter
Blower
Downflow

Type A1 Cabinet with canopy

canopy
Type A1 Cabinet with canopy
30% Exhaust air
Exhaust
Exhaust

air
30% Exhaust
Type Inflow
A1 Cabinet with canopy
Inflow
Blower 30% Exhaust air
Exhaust
Blower

(: WHO. Laboratory biosafety manual 2004) (: Eagleson institute 2004)

grille
Inflow institute 2004)
(:FrontInflow
WHO.
manual
: Eagleson
Laboratory
17 biosafety

class2004)
II (Airflow
Class II Type A1

(Supply
17

class
II

Airflow

Class
II Type
A1II (Supply
air
Split
air 17Split

II Front
grille (
Class
Room
air
)

class
Airflow

TypeA1
Blower
Rear

air

grille
Rear

air
Exhaust
filter
air grille
(
Split

Front
(
Room
(Supply
Front
grille
Room
Blower
)
grille

)
Supply
filter grille
30%

70%
Supply
filter

)
Rear

filter
Exhaust
Supply
filter

2004)
30%
(

institute
filter
70%2004)

Supply
(:Exhaust
WHO.
Laboratory
biosafety
manual
:
Eagleson
filter
30%Supply
filter

70%
Supply

filter
)
Type A2 Cabinet with canopy
)
17 Type
A2A2
classCabinet
II
Type
Cabinet
with Airflow
canopy Class II Type A1
with
canopy
Exhaust air
air )
Exhaust(30%
(Supply air Split Front grille
Room
Rear grille Blower
Exhaust
30%

Exhaust filter
Exhaust air
Supply filter 30% 70% Supply filter )
Blower
Type A2Inflow
Cabinet with canopy
Inflow
Exhaust
Blower

30% Exhaust air

(: Eagleson Institute 2004)

Inflow
18 (
Airflow
Class2004)
II Type A2
: Eagleson
Institute
(Supply filter 30%
70%Class
IIType
SupplyA2 filter )
18
Airflow
(Supply filter 30%

70%

Supply filter )
18 Airflow Class
Type B1II Type A2
(Supply filter 30%
air70% Type
B1 Supply filter )
70%
Exhaust
(

:
Eagleson
Institute
70% Exhaust air
(: 70%
Eagleson
Institute2004)
2004)
Exhaust
air
18 Airflow Class II Type A2 Exhaust 70% Exhaust air
(Supply filter 30% 70% Supply filter Exhaust
)
70% Exhaust air

18

Type
B1
Type Inflow
B1
Inflow

Blower 70% Exhaust air

Exhaust
Blower

(: www.scientistsolutions.com)
(: Eagleson Institute 2004)
Inflow
(: www.scientistsolutions.com)
: Eagleson
19 Airflow (Class
II Type B1Institute 2004)
(Supply air
Airflow
Frontgrille
B1 Rear grille 70%

19
Class30%II Type
Blower
70% (Supply
( ) air

30%

70%
Supply filter)
Front grille 30%
Rear
grille
70% ( ) / 30% Supply filter)
21
(: www.scientistsolutions.com)
(: Eagleson Institute 2004)
21
19 Airflow Class II Type B1
(Supply air
Front
grille
30%Class
IIType
Rear B1grille 70%
19
Airflow
70%
(

30%

Supply
(Supply air Front grille 30%
Rear grille
70%filter)
70% () / 30% Supply
21 filter)

 BSC
Type
B2 B2B2
Type Type
B2
Type
Exhaust
100%100%
Exhaust
air air
100%
Exhaust
air
Exhaust
SupplySupply
Supply Exhaust
Exhaust

BlowerBlower
Blower
InflowInflow
Inflow

Institute
(:(
(:(
Institute
2004)2004)
(www.scientistsolutions.com)
:: www.scientistsolutions.com)
www.scientistsolutions.com)
(Eagleson
:: Eagleson
Eagleson
Institute
2004)

20

Airflow

Class
II
Type
B2
20
Airflow

ClassClass
II Type
B2 B2

20
Airflow
II Type
(
Airflow

Supply
B2

air 20air

Class
IIair Type
(
RoomRoom

Supply
air

(
Room
air

Supply
air

Room
air

((
Room
air air
)

/

100%)100%)
Room

)

100%)

Room
44 Type

!
BSC
Class

!

(
airClass
II IIII 4
Supply
air

( Room air
BSC
BSC
Class
Type
Type

Class
II

(Type)

A1,
A2,
B1

II

4
(Type)
/
A1,
A2, A2,
B1

B2 B2
Class
(Type)

A1,
B1
B2
) Class

II
4

100%)

BSC Class

4
Type

44Type

!
II

4 Type

Type

A1,

A2,

B2

Class II

(Type)
B1

Type
A1

A2

A1

A2

Type
A1
A2 A2

A1

A2 A2

Type

A1

A1

(Canopy)

(Canopy)

(Canopy)

4
Type

(Hardduct)

Canopy

(Hardduct)
Canopy

(Hardduct)
Canopy


Type

Type
A1

A2

A1 A2

A2

Type

A1 A1

Type
B1 B1B1

Type
Type
A1
A2
A2

Type

B2
B2

(Canopy)

B2

- (Hardduct)
Canopy
Type A1 A2
Class
IIII
- - Class
II
A2 A2
Class
A1,
A1,
A1,
A2
Type B1 B2

A1,

A1,
A2 A2

A1,
A2

(Building
exhaust
system)

(Building
exhaust
system)

(Building
exhaust
system)
Class II
A1,
A2
(Canopy

connection)

(Canopy
connection)

A1,
A2
(Canopy

connection)

(Housing)

Exhaust
filter

(Building

(Housing)

Exhaust

system)
(Housing)
Exhaust

exhaust

filter
filter

22 22
22 (Housing) Exhaust filter
(Canopy connection)

19


Type A2
A2 Cabinet
Cabinet with
Type
with canopy
canopy
Exhaust
Blower
Inflow

(: Eagleson Institute 2004)

21 21 Canopy
Canopy

(: Eagleson Institute 2004)

Canopy
Canopy
2222

Exhaust

Exhaust air
air



A1 A1

A2 A2

Canopy

Canopy

Positive
air pressure*
pressure*

Positive
airPositive

Positive air
A1 A1

pressureair

pressure

pressure** A2 Positive air pressure

Negative air
air pressure**

Positive
air
Negative Negative
air pressure**

A2

pressure Negative air pressure**
*
Positive air
pressure
*** Negative
Positive air
pressure

air pressure

** Negative air pressure

B2

class II B1,

(Hard-ducted) Exhaust air
class II B1, B2

(Hard-ducted) Exhaust air

Exhaust air

B2 Negative air pressure

Negative
air pressure
B1

Exhaust air
Negative air pressure B1 B2 Negative air
pressure

23

20

 BSC

Airflow 4

Exhaust air Re-circulating air

BSC (Type) Exhaust air Type A1, A2
B1 (Re-Circulating air) B2
1
1 Re-circulating air, Exhaust air



(Re-circulating air) (Exhaust air) Lab
A1
70%
30%
-



- (



Canopy)
A2
70%
30%
-



- (



Canopy)
B1
30%
70%

B2
0
100%


(Airflow velocity)

(Inflow velocity) (Downflow velocity)
Specification
(Manual usage) 2
(Air velocity)
2 (Air velocity) Exhaust air
Recirculation air Exhaust air
Type of BSC
A1
A2
B1
B2

Inflow/Intake Velocity Downflow Velocity Re-circulation Exhaust

75fpm*/0.38mps**
75fpm/0.25-0.5mps
70%
30%
100fpm/0.51mps
50fpm/0.25-0.5mps
70%
30%
100fpm/0.51mps
50fpm/0.25-0.5mps
30%
70%
100fpm/0.51mps
80fpm/0.25-0.5mps
0%
100%

*fpm: Feet per minute ** mps: Meters per second

BSC HEPA

21




HEPA filter


A1 A2 (Volatile chemicals) Acetone,
Chloroform, Methanol Alcohol A1, A2
(Re-circulate air)

A2 (), B1 B2

A2 B1
A2 B1 Airflow Rear grille Exhaust
30% 70% 2
(Minute amount)

B2 (Small amount) A2 B1
(Re-circulating air) (100%)

B2

22

 BSC

BSC Class III

BSC Class
III
: Class III

: Class
III

(Gas-tight)
(Gas-tight)

BSC
Class
II

Gas-tight

Class
II BSC

Gas-tight

Double-doored

Double-doored
autoclave autoclave

(Dunk
tank)

(Dunktank)

Rack

Rack Incubator

HEPA
filter
:

HEPA
filter

Supply
air

Exhaust
air

Exhaust air
Incubator
HEPA filter 2HEPA
filter : HEPA filter Supply air Exhaust air

Airflow
:
filter
2
Exhaust air
HEPA
Airflow
:

23 Class III (: /Uvm.edu/safety/lab)

26

23

Airflow !

Inflow airflow (Front opening)
(Operator protection)

Down flow HEPA filter


Exhaust airflow HEPA filter

Airflow

Class I - III Class II 4 Type
Airflow
...

BSC
HEPA filter, Blower Laminar flow Airflow
Class II
A1, A2, B1, B2

24

25

 BSC

(Proper Selection)

BSC 3 Class
(Product protection), (Personnel protection)
(Environment
protection)



1.
Class I
Class II

2. (Risk group) Risk group 1, 2,
3 4 Risk group Risk group 3
Risk group 1 2

3.
(Volatile toxic chemicals)
BSC

A1
B1

26

A2
B2

?
Risk group? 3P?

 BSC
3

(types of protection)
Personnel protection for risk groups 1-3
Personnel protection for risk groups 4
Product protection
(Volatile chemical)
(minute amount)
(small amount)

BSC
class I, class II (A1,A2,B1,B2), class III
class III
class I, class II ( positive suit
laboratory)
class II
class II B1, class II A2 (A2
)
class I, II B2, class III

B1 B2
3.

(
28) BSC (Duct)

(Exhaust)
..

-

.....
-

B2

Exhaust
air 100%

(tabletop
certrifuge)

vortex
(A1, A2,mixer
B1

B2)

Airflow

airflow

...

27

 BSC

BSC

1. (Location)

(Inflow air) 0.38

( BSC ) BSC
BSC
30
30-35 (Exhaust filter)
Exhaust filter

2. (Personnal)

(Standard microbiology practice)
Inflow air



1
()

3. (Material placement)

,
Tissue culture flask

!
(Front inflow grille)

(Rare grille)
28

 BSC
(tabletop
certrifuge) vortex mixer Airflow
airflow

70% Alcohol




Inflow air

4.

BSC

5. Ultraviolet light (UV)

UV UV
BSC UV


UV UV

UV


UV
UV
40 w/cm2
UV

BSC

29

- UV
UV 70% Alcohol

4


UV

UVUV BSC
- UV

UV UV

BSC

UV


70%
Alcohol

UV


70%4 Alcohol
4


...
6.

UV(Open
flames)

BSC

UV

UV

BSC

(Bunsen)
BSC

BSC
UV

BSC
(Open
6.

(Open
flames)
flames)

(Bunsen)

?
flames)

6.

(Open

(Air flow pattern)

BSC
(Bunsen)

(Open
BSC

flames)

?
(Openflames)

(Air(Airflow

flowpattern)
pattern)

BSC

Loop

Sterile
technique

(Sterile)

BSC
BSC

Sterile
technique

(Sterile)

Sterile
Micro-burner
Electric
furnaces

technique
(Sterile)
LoopLoop

Micro-burner
furnaces

furnaces

Electric
Micro-burner
Electric

AIR

AIR GAS
GAS

24

24

:
commons.wikimedia.org)
(: commons.wikimedia.org)

7. (Spills) BSC

UV

UV
BSC

1.

2. 0.05% Clorox ( 5.25% Clorox 1:100)35
35 Clorox
30
1:10 (0.5% Clorox)

3. 70% Alcohol (
UV UV 30 )

BSC ...

UV

30

 BSC

1.

2. 3-5

3. 70% Alcohol

4. 70% Alcohol

5. (Steriled test tube, Tip, Plate )
( )

1. / 1

2. (Front grille) (Rear grille)

3. 4 Front grille

4.
(Cabinet airflow)

5.


6. Airflow

7. (Clean) (Dirty)
Pipette, Test tube, Flask, Media Pipetter
seed virus Pipette

8.

9. Supply airflow

10. Plate/dish Plate Plate Tube/flask

(Downward air)

11. Airflow Centrifuge, Blender
1/3

31










Clorox

1. 70% Alcohol
2. 70% Alcohol
3. ()
4. /
5. 70% Alcohol
6. 3-5
7. (Sash)
8. () Autoclave
Autoclave
9.
10.

BSC

32

33

 (Certification)
BSC

(Certification) Certify Physical testing

1 (Certifier)

1. Downflow velocity test ( )

2. Inflow velocity test ( )

3. HEPA filter leak test ( HEPA filter)

4. Airflow smokes pattern test (Smoke test) ()

5. Cabinet leak test ( A1 ) ()




1. lighting intensity test ()

2. Vibration test ()

3. Noice lever test ()

4. UV light intensity check ()

5. Electrical leakage ()

6. Alarm, Interlocks, Negative pressure test

Certifier (Accredit)



BSC

34

(Certification)

1. Downflow velocity test

! !

1.
Downflow velocity test
Type
B1
Class II,
1. Downflow
velocity

1. Downflow velocity
test

test

!
1. Downflow velocity test ClassClass
II, Type
B1

II, Type
B1
1. Downflow
velocity test

Class
II,
Type
B1

Class II, Type B1
Class II, Type B1

HEPA Filtered Air

HEPAContaminated
Filtered Air Worksurface Air
Contaminated
Contaminated
Room AirAirAir
HEPAWorksurface
Contaminated
RoomFiltered
Air
FilteredWorksurface
Air
Contaminated
Air
25 Downflow HEPA
(Filtered
: HEPA
www.nuaire.com)
Air
ContaminatedRoom
Worksurface
Air
25 Downflow (Contaminated
Contaminated
: www.nuaire.com)
Air
Worksurface
Air
Contaminated Room Air
Contaminated Room Air

25
Downflow
( : : www.nuaire.com)
www.nuaire.com)
25
Downflow
25
Downflow
( (: www.nuaire.com)

26 Thermal anemometer Probe

26 (
:
Thermal anemometer Probe

www.afrims.com)
( : www.extech.com)
( : www.afrims.com)
( : www.extech.com)
26 Thermal anemometer Probe
26 (
Thermal anemometer Probe (

: www.extech.com)
: www.afrims.com)
( : within
www.extech.com)
1.1 (
: www.afrims.com)
Thermal Anemometer
calibration due

1.1 Thermal Anemometer within calibration due

dateStand
Stand& &Clamp
Clamp
Probe
Probe
date

1.1

Thermal
Anemometer
within
calibration
due & Clamp

1.1
1.2
Thermal
within
due

26

Anemometer
Probe
date

due

anemometer

calibration

6 6within

Stand

Thermal
Thermal
Anemometer
calibration
1.2(1.1

date
: www.afrims.com)
(Probe
www.extech.com)
date
&Stand
: Probe
Probe
(Grid)

Stand

Clamp
& Clamp

(Grid)

1.3

4 46

Sash
1.2
1.2

1.2 1.3

Probe

Sash
(Grid)
Probe

(Grid)

1.4Downflow
Downflow

(Grid)
test

1.3 1.1Probe1.4

test
4 Thermal

Sash
1.3

Anemometer
within
calibration
due
1.3

Probe

Sash

Sash

Probe

41

1.4 Downflow test 1.4

date
Downflow
Stand
Clamp
1.4 Downflow
test test
&

Probe 41

41
1.2 6 41

(Grid)
4in(10cm)
6in(15cm) 6in(15cm)
1.3
Probe 4 4 Sash
6in(15cm)
1.4 Downflow test

41
27 Grid Probe Downflow
( : www.afrims.com)

2. Inflow velocity test

Class II, Type B1

35

 !
velocity test

27
Grid

Probe
Downflow

27
Grid

Probe
Downflow
( : (
www.afrims.com)
: www.afrims.com)

ClassInflow
II, Type
B1 test
2. Inflow
velocity
test

velocity

2. Inflow velocity2.test

Class II, Class
Type II,
B1 Type B1

Class II, Type B1

HEPA Filtered
HEPAAir Filtered Air
ContaminatedContaminated
WorksurfaceWorksurface
Air
Air
HEPA
Filtered
ContaminatedContaminated
Room Air
Air Room Air

Inflow Inflow

Contaminated Worksurface Air

Contaminated Room Air

25 Downflow ( : www.nuaire.com)

Direct Inflow
Direct Inflow
Measurement
Measurement
(DIM) with(DIM)
hoodwith hood

( : (
info.baker.com)
( : (
www.globalspec.com)
: info.baker.com)
: www.globalspec.com)
28

Inflow
Dim

28
Inflow

withDimhood
with
hood

42 velocity 2
42Inflow
Inflow velocity 2 : 1. Primary method 2. Alternative method

1. Primary method 2. Alternative method

Primary method

Primary
Thermalmethod
anemometer
with
Probehood

BSC Seal
1. DIM

: www.afrims.com)
(
: www.extech.com)
hood


1. DIMwith
BSC Seal
2. 5
5

2.
3.Anemometer

(Volumetricwithin
rate, calibration
Q) = (Air velocity,due
V x Cross sectional area, A)
Thermal
3.
(Valumetric
flowflowrate,
Q)
= (Air velocity,
V x Cross sectional area, A)
Q = VA

ate Stand
& Clamp Probe Q = VA
Alternative
method
1 6 1
Alternative
method


1. 1.Thermoanemometer

Thermoanemometer

Grid)

3
Probe

Sash
2.

1 1

2.

ownflow test

41
29 ( : www.afrims.com)

36

Alternative method 2

1. Anemometer Exhaust filter


29 ( : www.afrims.com)

(Certification)

Alternative
2
Alternative method
method
2

1. Anemometer

Exhaust
filter filter
1. Anemometer

Exhaust

30 Inflow velocity Exhaust filter method ( : www.afrims.com)

43

3. Airflow smoke patterns test

smoke patterns
test

test)

3.Airflow

(Downward

(Downward test)
3. -Airflow
smoke patterns test

-
4

(Downward test)
4

Smoke
Smoke test
test Airflow
Airflow
3131
( :: www.afrims.com)
www.afrims.com)
(
31 Smoke test Airflow
-

( : www.afrims.com)

(View screen
retention

(View
test)
6

screen
screen
retention test)
-

(View
retention
6
1
test)1

6
1

32
Smoke
Smoke test
test

32
(

:
www.afrims.com)
: www.afrims.com)
32 (Smoke
test
-
(Inflow)
( :www.afrims.com)

opening
(Inflow)

edge
retention

(Work

(Inflow)

test)

(Work opening
test)
edge
retention

(Work opening edge retention test)

33
Smoke
Smoke test
test

Inflow
Inflow air
air
33
( :: www.youtube.com)
www.youtube.com)
(
33 Smoke test Inflow air
-


(
: www.youtube.com)

44 -

44

37

(Work opening edge retention test)

34 Smoke test
( : www.afrims.com)
34 Smoke test
4. HEPA filter leak test (
()
: www.afrims.com)
4. HEPA
filter

)
leak
34 test
Smoke
test

HEPAfilterfilter

Aerosol

DOP
4.HEPA
leak
test
(

)
(
: www.afrims.com)

HEPA filter

Aerosol
DOP
Aerosol
PAO Aerosol

BSC
Aerosol
PAOHEPA

BSC

filter

DOP
4. HEPA
filter
leak
test

generator
(

)
generator
filter
Aerosol

Aerosol

Aerosol

Aerosol
Aerosol
Filter

photometer

BSC

Aerosol
generator

PAO
HEPA
photometer

filter
DOP

BSC

generator
photometer

Aerosol
Filter

PAOAerosol

Aerosol

photometer

Filter
Aerosol
Aerosol

Aerosol photometer ( : www.hellotrade.com) Aerosol generator ( : www.atitest.com)

35 HEPA leak test
Aerosol photometer ( : www.hellotrade.com) Aerosol generator ( : www.atitest.com)
35 HEPA leak test
Aerosol photometer ( : www.hellotrade.com) Aerosol generator ( : www.atitest.com)
35 HEPA leak test

36 Aerosol photometer HEPA

( : escoglobal.com)
36 Aerosol photometer HEPA
5. Cabinet integrity

( : escoglobal.com)
36 Aerosol photometer HEPA

A1
Bubble
test

5.Cabinet
integrity Class

( II: escoglobal.com)

Class
4 IIA1

integrity

Bubble
test

5. Cabinet

5.
Cabinet

Bubble

integrity

II4A1

test

Class

Class II A1
Bubble

4
4

test

45

45

Certify 45

38

BSC Decontamination ( BSC)

BSC Decontamination BSC
(Fumigation) HEPA filter
(Certification)

1. (Certification)
2.
3. HEPA filter, (Motor/Blower)
4.

Formaldehyde Paraformaldehyde

1. : , , Hot plate/Gas generator (
Remote control), , & , (Respirator)

2. (Acess opening) (Exhaust openings)

3. Paraformaldehyde (: 0.3 x Cubic feet = )
0.8%

4. Heat Hot plate Formaldehyde

5. 21C 60-85% RH

6. 6 (Contact time)

7. Formaldehyde (Neutralization) Ammonium Bicarbonate

8. (Ventilation) 1

9. (Verification) Sport strip 7
10. 70% Alcohol


37% Formalin, Hydrogen peroxide, Ethylene Oxide Chlorine dioxide

39

 Chlorine dioxide
Decontamination
Decontamination Decon
Decontamination
Decon

Decontamination
Decon

Decon

37 Decon BSC Hydrogen peroxide vapor

( : bbs.sciencenet.cn)

...

1. Horizontal laminar flow clean benches BSC

47
Horizontal laminar flow clean benches BSC
Horizontal laminar flow clean benches Clean benches BSC

Clean bench HEPA filter

(Product production)

Sterile
equipment Electronic devices

(Sash/window) (Grille)

Clean bench Tissue culture



Vertical laminar clean benches (Clean benches) BSC Clean
bench

Horizontal clean bench
(Sash) BSC Grille

Herizontal clean bench
Sterile normal saline

Tissue culture Horizontal cleon
benches Sach Vertical clean benches
BSC
40


2. BSC Chemical fume hood

Class II BSC (Product protection)
(Personal protection)
(Environment protection) HEPA filter

Chemical fume hood (Personnel protection)
Chemical fume hood
HEPA filter
3. Biological Safety Cabinet (BSC) Class I, II III

Cass I (Personnel & Environment protection)

Cass II (Product, Personnel & Environment
protection)

Cass III (Product, Personnel & Environment
protection)
4. HEPA filter ?

HEPA filter

HEPA filter 0.3 99.97%

HEPA filter 0.3 99.99%
5. HEPA filter ?

HEPA filter
6. (Certification)

Specification
(Certifier) HEPA filter

Filter
1
7. HEPA filter leak test (Test aerosol) 0.3 ?

HEPA 0.3
0.03% 0.3
0.3
0.03%
8. (Standard Microbiological Practice)

Contact lens Lab

41

(Aerosol)

9. (Laboratary technique)

(Laboratary technique) Good laboratary technique



9.1
(Label)


9.2 Rack



9.4

9.5
BSC

9.6
(Micrological transfer loops) 2-3
6
Electric microincinerator Loops Loops

Autoclave
Sterile tape


9.7
5


42


/




9.8

Pasteur pipette

9.9





Autoclave
Blood clot
Autoclave (Incineration)


9.10
BSC

9.11 Hypochlorite Hypochlorite
(Freshly prepare) Chlorine 1 g/1 lit

9.12
(Pipet aids)





BSC

43





9.13 Biological safety cabinet (BSC)


BSC

BSC
(Glass viewing panel)

(Rear grille)
HEPA
filter Electric microincinerator

(Air grille)


(Cabinet fan) 5
BSC

9.14 (Centrifuge)

(Bucket)

(Specimen container) (Centrifuge tube)

Bucket
70% Alcohol Bucket Bucket
Rotor bucket
44



9.15 Homogenizer, Shaker, Blender Sonicator
(Homogenizer)
Blender
BSC

9.16


(Label)
Label Autoclave

10. NSF 49

Standard No. 49 BSC The National Sanitation
Foundation International (NSF)
NSF BSC 1 Type
Size Biological test, Physical test/
Certification, Structural test Clean ability test.

American Standard NSF 49

European Standard EN 12469

Australian Standard AS 2252

Japanese Standard JIS 3800

11. Decontamination Disinfection, Antisepsis, Sterilization
Sanitization

Disinfection
(Physical treatment)
/ (Disinfectant) Alcohol, Lysol, Bleach


Antisepsis

/ (Antiseptic) Hydrogen peroxide, Alcohol, Boricacid

Sterilization
(Boil) (Autoclave) (Heat)

Sanitization
(Sanitizer)

45

P1, P1,
P3 P3

P2

Decontamination (
(

P2
Physical
containment)
containment)
(Fumigation)
Physical

BSC

(Certification)
BSL1,
BSL2,BSL2,
BSL3BSL3

BSL4B

BSL1,
HEPA filter

12. Biosafety levels (BSL)

BSL
3-4 3-4

BSL

4

BSL1

(Lab
supervisor)

(Lab
supervisor)

practice)


(Standard
microbiogical

BSL1

12.1

(BSL)

12.1

BSL1

P2

P3

P1,

Physical containment)

BSL1,

BSL2,
BSL3 BSL4

3-4

BSL

(Sterile
technique)

(Sterile

technique)

(Lab supervisor)

(Facillity
design)
BSL BSL
1

(Facillity
design)
1
12.1 BSL1

(Sterile
technique)

(Facillity design) BSL 1

58 58

38 38

BSL
1BSL rd1
(: (WHO.
Laboratory
biosafte
manual.
3 2004)
: WHO.
Laboratory
biosafte
manual.
3 rd 2004)

12.2 BSL2

BSL2 BSL1







(Aerosol or Splashes)
(Biological safety cabinet, BSC) BSC Class I Class II

46

 BSC

Eyewash

(Facility design) BSL 2
(Lockable doors)




BSL3
12.3
BSC
(Aerosol
39 BSL 2

route) (:WHO.
Laboratory
biosafte
manual.
3rd 2004)

Eyewash


(Personnel protective equipment)
BSC 2 BSC Class I Class II

(Facility design) BSL 3

12.3 BSL3
BSL1

BSL2

route)

12.3
BSL3

(Aerosol

(Aerosol

(Personnel

BSC inward
airflow)

equipment)

(Directional
ute)

protective

2

Class
I
Class

BSC
II

(Personnel protective equipment)

(Facility
design)I
BSL Class
3

BSC
2
BSC Class
II
BSL

BSL1

(Facility design)
3
BSL2

BSL2

BSL1




2

2

(Directional inward airflow)

(Directional inward airflow)

60

40 BSL III
(: WHO. Laboratory biosafte manual. 3 rd 2004)

40 BSL III
(: WHO. Laboratory biosafte manual. 3 rd 2004)

60

41 BSL III
( : En.wikipedia.org)

12.4 BSL4

41 BSL III
( : En.wikipedia.org)

47

(Primary and secondary containment functions)

-
(Containment equipment) /
-
(Laboratory director)
-

and


(Primary

secondary containment functions)
-

(Containment
equipment) /
III

-
BSC
Class
(Laboratory
director)
(Facility design) BSL 4



BSL3
:

Class III Glove box laboratory

(Possitive
pressure suit/Suit labor

BSC Class III

(Facility design) BSL 4
BSL3 :
Class III Glove box laboratory
(Possitive pressure suit/Suit labaratory)

42 Positive suit
( : bepast.org)

Reference

1. U.S. Department of Health and Human Services, Public Health Service, Centers for Disease
Control and Prevention and National Institutes of Health. Biosafety in microbiological
and biomedical laboratories (BMBL). 5th edition. Washington: Primary containment for
biohazards selection, installation and use of biological safety cabinets (Appendix A) and
standard microbiological practice; 2007.
2. World Health Organization (WHO). loboratory Biosafety Manual. 3rd edition. Jeneva: Biological
safety cabinets & laboratory technique; 2004.
3. Esco Micro Pte Ltd. A guide to biosafety & biosafety cabinets. Singapore: Biohazard safety
cabinet selection; 2005.
4. Safety cabinet technology seminar, 1-2 November 2004. Eagleson institute, Sanford, ME, USA
5. Biological Safety Cabinet Certification Training Course. 1-5 October 2012 AFRIMS-National
Institute of Animal Health, Bangkok.

48

49