What are the significant differences between the requirements for negative pressure rooms and C-PECs of <797> and <800>?
USP<800> is meant to work in conjunction with <797>, so a lot of the regulations are harmonious. There are a few exceptions, and expect <797> to be updated with these new changes shortly. USP<800> states that the negative pressure room has to be between 0.01 and 0.03 inches of water column more negative than the surrounding areas. <797> didn’t have an upper limit. C-PECs now have to be located within a negative pressure room for all HD work. This does not have to be a CACI. The last difference is that all HD work needs to be done in a C-PEC.
• I need to decontaminate my Cultivo without using the onboard vaporized Hydrogen Peroxide program. What chemicals can I use?
Certain chemicals have been shown to be harmful to the RH sensor. This will cause an inaccuracy in the ability of the RH sensor to identify the true internal humidity. Do NOT use any chlorine-based chemicals (e.g. bleach). Baker recommends using the following for surface decontamination:
Diluted Hydrogen Peroxide
After cleaning, a rinse with sterile distilled water is recommended. Also, allow all fumes to exit the incubator before closing the door and resuming operation.
Does the BioChemGARD Class II Type B2 installation require a specific type of exhaust valve in order to operate in ReadySafe mode? (viewscreen closed)
~~No, the BioChemGARD 401 and 601 does not require a special exhaust control valve in order to operate the ReadySafe mode feature.
The ReadySafe feature is standard on all BCG401 and BCG601 biosafety cabinets and activates by lowering the viewscreen to its fully closed position. Once the viewscreen is closed, the BCG supply fan speed is automatically decreased and turns the fluorescent lamp off, reducing the biosafety cabinets total energy consumption. In the ReadySafe mode, the facility exhaust system requirements will not change, pulling the required air through the cabinets bypass armrest and into the front worksurface perforation maintaining the cabinets personnel and product protection characteristics.
For additional energy savings, the customer may choose to equip the laboratory with a multi-position exhaust control valve to reduce the volume of conditioned lab air exhausted from the space. This is not a requirement for the BCG401 and BCG601 but rather an optional configuration supplying dry Form C contacts to communicate with the facility exhaust valve. If this option is being considered, room air balance and pressurization must be taken into account.
Is it recommended to have more than one person working in a biosafety cabinet at one time?
The Baker Company does not recommend more than one user working through the front access opening of any Class II biosafety cabinet. Although our Class II products have undergone a rigorous test evaluation using microbial spores to ensure personnel, product, and cross contamination protection, these tests are performed in an ‘at rest’ condition as stated in NSF Standard 49.1 When used properly, Class II BSCs have been shown to be highly effective in the reduction of laboratory-acquired infections and/or hazardous drug exposure, the protection of product from outside contaminants, and protection of cultures from cross-contamination. Currently there are no ‘use restrictions’ within the biosafety industry regarding simultaneous use; therefore, a risk assessment should be performed by the individual most familiar with the product being considered for use. A biosafety cabinet is a valuable supplement to, but not a replacement for, good laboratory technique and safe practice.
Does NSF provide a document that offers guidance on biosafety cabinet selection, installation, lifespan, or decommissioning?
Yes, NSF has published Annex E, which provides this type of guidance.
Do USP <797> or USP <800> provide guidance about which side of the anteroom the sink should be located? Dirty side as people are entering and donning booties, or clean side where we are diminishing the particulate load toward the clean room?
Neither USP <797> nor <800> specifically calls out a sink location within an ante room, only that it is a critical component in reducing microbial contamination from entering the buffer area. For most applications that we have seen, the location of the sink is on the clean side of the anteroom or on the opposite line of demarcation used for hand and forearm washing after head, facial and booties are donned. It’s important to remember that sinks can be a major source of contamination; therefore, strict emphasis on environmental monitoring and gowning procedures must be made. We located a similar question on the USP797.org website here: http://www.usp797.org/QA-RJLG6.htm
We do recommend, however, that our customers employ their own risk assessment, relative to applicable regulatory guidelines, facility requirements, and standard operating procedures in order to derive at the right decision for them.
Does the fan run continuously, even when you open the door, or will the fan shut off when you open the door?
Yes, the fan will be disabled when there is a door opening. Air from the outside will influence the chamber’s ability to maintain a warm, humid environment essential to cells and cell growth. If the fan were to run when the door is open, moisture and heat will escape and be blown out of the unit, thereby increasing the amount of time it takes to recover the environment. When the door closes and the fan is turned back on, Cultivo will return to better than ISO Class 4 (Class 10) conditions within 60 seconds.
Will the down flow of air and the rate at which your incubator moves air throughout the chamber negatively impact my cultures?
The capacity of the unit to maintain plate and fluid moisture has not been shown to pose a problem within the Cultivo design. The design of petri dishes and T-flasks, is supposed to allow gas exchange into the culture without drying. Turbid air within the chamber and any upward airflow would actually have a much larger impact (when dishes are covered and capped, air can enter in more easily from the bottom than from the top). Cultivo’s uniform down-flow air provides ISO Class 4 Air to the chamber at a rate which will not impair efficient gas exchange or dry out media / cultures. Please review our test reports on air cleanliness and cell growth.
In the Cultivo Ultra and Ultra Plus, which feature ultrasonic humidification, is the humidification done before the HEPA filter or after?
Since we know the water reservoir to be the most likely source of contamination within most incubators, the HEPA filter has been placed after the RH generator.
What is the difference between the two different methods of humidification you supply (Evaporative vs. Ultrasonic)?
The Cultivo with evaporative humidification relies on water evaporating from a pan and into the airstream. This method of humidification is capable of providing a 10 minute recovery to our high end set point (92%, +/- 3%). The Cultivo Ultra with ultrasonic humidification (nebulizer) uses ultrasonic vibrations to aerosolize water which is then readily evaporated into the air. This system provides 20% faster humidity recovery and does not require the user to access the chamber in order to refill/replenish/empty a water pan, thereby reducing the likelihood for contaminants to enter the system from the outside.