Using your Biosafety Cabinet to protect you and your work Name: - - PowerPoint PPT Presentation

Using your Biosafety Cabinet to protect you and your work Name: Bernard Schwartz Title: Controlled Environment Product Specialist Date: July 25, 2017

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Agenda

• 1. A2 BSC
• 2. HEPA Filter
• 3. Aseptic Technique – General
• 4. Organize workflow
• 5. Before Starting
• 6. Starting Work
• 7. Working Tips
• 8. Universal Precautions
• 9. General Precautions
• 10. Waste
• 11. Spills
• 12. Questions

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Class II, Type A2 BSC

• Class II, Type A2 BSC

Airflow Diagram

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HEPA Filter

0.000001% 0.000010% 0.000100% 0.001000% 0.010000% 0.100000% 0.01 0.10 1.00 Percent Penetration as a percentage of upstream Particle size in micrometers

Typical HEPA Filter Efficiency expressed as percent penetration

Bacteria Virus

0.000001% 0.000010% 0.000100% 0.001000% 0.010000% 0.100000% 0.01 0.10 1.00 Percent Penetration as a percentage of upstream Particle size in micrometers

Typical HEPA Filter Efficiency expressed as percent penetration

Bacteria Virus

HEPA Filter Efficiency

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Aseptic Technique

This facility manufactures products for clinical use These products must meet a number of requirements, one of which is that they are sterile In order to produce sterile components it is necessary to follow aseptic technique

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Aseptic Technique

Use of aseptic technique protects the product and the operator from exposure to infectious agents and contaminants The technique is a way of working that provides this protection The following slides provide basic information on aseptic technique

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Aseptic Technique

The idea behind aseptic technique is to PREVENT contamination It is not possible to “clean up” an already contaminated product by using aseptic technique In order to maintain sterility, it is necessary to start with a sterile product & handle it in an aseptic manner

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Organize your work

How to organize your supplies and work in the biosafety cabinet

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Organize Workflow

• From the US CDC/NIH “Primary Containment for

Biohazards: Selection, Installation and Use of Biological Safety Cabinets” 3rd Edition

Clean (for right handed user) Dirty

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Tx Tech

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Caution with waste containers

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Tx Tech

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Is this on?

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Before Starting

Collect all of the reagents and materials that you will need onto a workspace close to the biological safety cabinet Make sure that you have a biohazard trash container, sharps container (if appropriate) and waste trap available – we recommend that waste traps be located on the floor OUTSIDE the BSC Make sure that you have disinfectant spray bottles containing 70% ethanol available

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Before Starting

Make sure that you are wearing the required personal protective equipment

• 1. Lab coat – Replace between products
• 2. Gloves
• 3. Eye Protection if necessary
• 4. Sleeve Covers - (when working with more than
• ne product)

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Before Starting

Use alcohol to clean gloves before using the BSC !!!!!!!

Minor Clean - Wipe down the working surface in the BSC with 70% ethanol and leave to dry

Disinfect any equipment e.g. racks, before placing them into the BSC

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Use of Alcohol

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Starting Work

When you start working, be sure that you work well inside the BSC and not directly above the front air vents Wrong !! Keep your nose and mouth away from the window opening

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Starting work

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Working Tips

Tighten the caps on culture flasks before moving them to the BSC. Wipe down culture flasks with spray disinfectant before bringing them into the BSC. Decontaminate your gloves using alcohol frequently during the culture procedure Be careful not to touch the necks of culture flasks when removing the caps. Be careful not to touch the inside of the cap Place the cap either face up, or face down on an alcohol wipe.

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Working Tips

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Finishing your Work

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UV

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Working Tips

Avoid extreme temperature changes A culture that becomes cloudy rapidly after incubating is probably contaminated All contaminated cultures should be sterility tested to identify the contaminating organism Cultures that contain white or gray threads, or cotton wool-like floating particles usually are contaminated with fungus or mold Fungus and mold is difficult to cure effectively

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Universal Precautions

All cell products should be regarded as potentially infectious Follow Universal Precautions when working with these products

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General Precautions

If you feel that you may have touched a sterile item – discard it and use a new one Clean up any spills immediately after decontaminating with disinfectant

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Waste

Discard old culture flasks into the biohazard trash after tightening the caps Disposable tips used with a Pipetman should be discarded into the waste container. DO NOT overfill waste containers

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How to handle a spill in the biosafety cabinet

• 1. Neutralize.
• 2. Disinfect with Clorox

Healthcare Bleach Germicidal Cleaner

• 3. Wipe everything removed with

70% ethanol.

• 4. Capture spill in appropriate

container via spill valve.

• 5. Wipe down spill tray
• 6. Wipe down work surface
• 7. Run UV decon

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Thermo Scientific Provides Tools You Can Use!

• At Thermo, we are committed to providing the best customer

experience before and after the sale

• Expertise and history continue to drive innovation
• Visit our website, www.thermoscientific.com/co2 to find:
• Information about new and existing features
• Interesting articles building on new discoveries
• Helpful Application Notes
• Informative Smart Notes
• And more!
• Have a question? Contact us!

Thank you for your attention!

Name: Bernard Schwartz Title: Controlled Environment Product Specialist Email: bernard.schwartz@thermofisher.com Phone: 615/714-2436