Saftey work in a biosafety cabinet class II

A Difference in Life Science

Safety work

in a

laminarflow

cabinet class II


- Function of a laminar flow 2 filters vs 3 filters

- How to work with pathogens

- Use and clean a cabinet from the start to the end

Safety work in a laminarflow cabinet class II

Laboratory devicesCabinet selection / types

• Clean Bench- Open fronted exhaust protective hood- Work protection only

• Class I- Open fronted exhaust protective hood- Operator / environmental protection

• Class II- Vertical laminar flow recirculating air cabinet- Operator / work / environmental protection

• Class III- Enclosed glove box with filtered air in and out- Operator / work / environmental protection



Laboratory devicesCabinet selection / types

• Clean Bench- Open fronted exhaust protective hood- Work protection only• Class I- Open fronted exhaust protective hood- Operator / environmental protection

• Class II- Vertical laminar flow recirculating air cabinet- Operator / work / environmental protection

• Class III- Enclosed glove box with filtered air in and out- Operator / work / environmental protection

HEPA filter


HEPA stands for High Efficiency Particulate Air filter.

They are normally 99.999% efficient against a particle size of 0.3µm. (Eurovent EN 13091:1999, EN1822-1 Class H-14 or higher.)


HEPA filter

HEPA Filter construction

Air flow

Filter Medium

Gasket

Filter Frame


Pre-filter Blower Plenum HEPA Work Area

Filter

Filter Technology - Airflow Principle


Filter Technology - Airflow Principle


Blower

Plenum

HEPA Filter

Work Area


Blower

HEPA Filter

Work Area

2-Filter Cabinet

Plenum

Bio Safety Cabinet Class II

- Air flow 70% recirculation 30% exhaust


Blower

HEPA Filter

Work Area

2-Filter Cabinet

Plenum

Pre-Filter




- Vertical laminar flow recirculating air cabinet- Air flow 70% recirculation 30% exhaust- Operator / work / environmental protection


2-Filter Cabinet 3-Filter Cabinet

Pre-Filter





Pre-Filter1st-Filter

H14

Effects by down flow speed

0.25 - 0.50m/s According EN 12469

• Too low speed (< 0.25 m/s) - less clean air at work place

• Too high speed (> 0.50 m/s) - turbulences at work table



Positive pressure

Working principleNegative - Positive pressure A2 Cabinet

Negative pressure

Blower

Filters


Common particle generation

Sitting or standing no movements

Sitting small movements of head, hands and arms

Siitting small movements of upper body, arms and feet

Standing up and sitting down

Slow walk (ca. 0.9 m/s)Normal walk (ca. 1.6 m/s)Fast walk (ca. 2.3m/s)

Particle size per m3 / Min (0.3 um) (0.5 um) 004 Mio 001 Mio

0 17 Mio 005 Mio

0 35 Mio 010 Mio

87Mio 026 Mio

175 Mio 052 Mio 262 Mio 073 Mio 350 Mio 105 Mio


- Function of a laminarflow 2 filters vs 3 filters

- How to work with pathogens



Biosafety Level

BSL 1 - Little or no riskBSL 2 - Moderate risk Prophylaxis and treatment availableBSL 3 - High risk. Prophylaxis and treatment if treated early enough available BSL 4 - Very high risk. No treatment available

Typical Examples:

Group 2 - Botulism, Legionella, E.coli strainsGroup 3 - Anthrax, Typhoid, HIV, Hepatitis, BSEGroup 4 - Lassa fever, Ebola, Green Monkey Disease


BSL 1 - Little or no riskBSL 2 - Moderate risk Prophylaxis and treatment availableBSL 3 - High risk. Prophylaxis and treatment if treated early enough available BSL 4 - Very high risk. No treatment available

Typical Examples:

Group 2 - Botulism, Legionella, E.coli strainsGroup 3 - Anthrax, Typhoid, HIV, Hepatitis, BSEGroup 4 - Lassa fever, Ebola, Green Monkey Disease


Biosafety Level

Working safely with pathogens

• Before bring in and take out - clean with ETOH 70%• Use 2 gloves together (specially with nanoparticle)

• Do not operate the cabinet if any of the alarms are activated• A bunsen burner should not be used in a cabinet at all• Do not use the cabinet as a storage area• Minimize disturbances to airflow barrier• Observe correct sash opening height (20 cm)• Observe surface – decontamination (periodically clean with ETOH 70%)

• The Cabinet must be controlled annually (down flow speed, leektesting of the hepa filter etc)




• Do not operate the cabinet if any of the alarms are activated• A bunsen burner should not be used in a cabinet at all• Do not use the cabinet as a storage area




• Do not operate the cabinet if any of the alarms are activated• A bunsen burner should not be used in a cabinet at all• Do not use the cabinet as a storage areaDo not use the cabinet as a storage area




• Do not operate the cabinet if any of the alarms are activated• A bunsen burner should not be used in a cabinet at all• Do not use the cabinet as a storage area• Minimize disturbances to airflow barrier• Observe correct sash opening height (20 cm)• Observe surface – decontamination (periodically clean with ETOH 70%)

• The Cabinet must be controlled annually (down flow speed, leektesting of the hepa filter etc)



- Function of a laminarflow 2 filter vs 3 filter

- How to work with nanoparticles and pathogens



Cabinet work procedures

Morning • Clean work table surface with 70% alcohol • Cabinet must run min. 15 min before working

During work day• Minimum stand by mode

Evening• Clean down work surface & spill tray with 70% alcohol• Cabinet must run 15 min after cleaning• Decontaminate with UV-light for min. 15 min• Cabinet can run on stand by mode during night



Morning • Clean work table surface with 70% alcohol• Cabinet must run min. 15 min before working






possible contaminated areas

Cabinet work procedures: Morning procedure – before first start



possible contaminated areas

Cabinet work procedures: Morning procedure – ready for work




Evening• Clean down work surface & spill tray with 70% alcohol• Cabinet must run 15 min after cleaning • Decontaminate with UV-light for min. 15 min• Cabinet can run on stand by mode during night




Still possible contaminated areas

Cabinet work procedures: Evening procedure



Still possible contaminated areas

Cabinet work procedures: Evening procedure – after cleaning below table top



Cabinet work procedures: Evening procedure – after last run


Thanks for your attetion


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Saftey work in a biosafety cabinet class II

Science

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