Using Biosafety Cabinets

Jim Hunter, Senior Project Manager, Labconco Author

11/16/2017 Added

356 Plays

Description

Those who use, intend to use, or select and purchase biosafety cabinets will find this colloquium particularly informative.

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[00:00:10.342]For Environmental Health and Safety
[00:00:12.676]I am Elizabeth, Betsy, Howe, representing them,
[00:00:16.261]and The Office of Research and Economic Development
[00:00:19.551]would like to welcome you all here today
[00:00:22.520]to the Laboratory Safety Colloquium.
[00:00:26.157]These colloquia are offered in recognition
[00:00:28.846]of the commitment by faculty and staff
[00:00:30.843]of the University of Nebraska Lincoln
[00:00:32.462]to assure a safe and healthy environment for teaching
[00:00:35.929]and carrying out your very meaningful research.
[00:00:38.467]Before we begin, I did mention the handouts,
[00:00:41.857]and we're not likely to have a tornado or anything,
[00:00:44.924]like that today, but if we should need to exit
[00:00:49.386]because of fire alarm, we do need to take the stairs
[00:00:52.990]which are to the left of the elevator over there.
[00:00:57.552]Today's topic is Using Biosafety Cabinets
[00:01:00.687]and due to some unforeseen circumstances,
[00:01:03.173]Labconco has substituted experts,
[00:01:07.069]so our presenter today, Jim Hunter,
[00:01:10.151]is a Senior Project Engineer with Labconco Corporation.
[00:01:14.323]Jim earned a Bachelor of Science,
[00:01:16.317]followed by a Master's Degree in Biological Science
[00:01:18.931]from Michigan State University,
[00:01:21.007]and then a second Master of Science
[00:01:23.092]in Immunology and Microbiology from
[00:01:25.622]Wayne State University College of Medicine.
[00:01:28.456]Jim has been with Labconco a number of years,
[00:01:31.407]and is an active author, lecturer, and researcher.
[00:01:34.878]He is responsible for new product development,
[00:01:37.209]and current product technical support
[00:01:39.758]for Class I and II biosafety cabinets,
[00:01:42.574]vertical and horizontal flow clean benches,
[00:01:45.440]PCR enclosures, ventilated animal enclosures,
[00:01:49.175]and ventilated forensics products.
[00:01:51.813]So Jim, welcome to UNL.
[00:01:54.141]Thank you very much.
[00:01:56.026]Thank you all for attending,
[00:01:56.859]I appreciate you taking time out of your day
[00:01:59.202]to take part in this.
[00:02:00.508]So we're gonna talk about biological safety cabinets,
[00:02:04.277]And compare them to other ventilating enclosures
[00:02:06.299]that might be in the lab and their differences.
[00:02:09.439]A little bit about how a safety cabinet operates,
[00:02:11.748]the mechanics of it.
[00:02:14.294]We'll talk about ergonomics
[00:02:15.491]because that's something that most users don't get right
[00:02:18.183]and as a result, either they or their work suffer for that.
[00:02:22.113]And then talk about, really, generic procedure,
[00:02:24.956]what should we do step-by-step
[00:02:26.526]when we're gonna work in the cabinet.
[00:02:28.295]And then we're gonna wrap it up
[00:02:29.912]with some audience participation.
[00:02:31.146]I've got some photos off the Internet
[00:02:33.335]of people misusing and abusing cabinets
[00:02:35.770]and hopefully at the end of this presentation
[00:02:37.635]you guys are gonna tell me what they're doing wrong, okay?
[00:02:41.249]So, you're gonna see a number of
[00:02:43.209]different ventilated enclosures in the lab,
[00:02:45.138]and these are just some that we make,
[00:02:46.774]by no means exhaustive.
[00:02:47.883]But if you look at these devices,
[00:02:49.839]a lot of them look very much alike,
[00:02:51.616]but I have four very different devices here.
[00:02:53.851]The first one on the left is a Clean Bench,
[00:02:56.999]next is a Ductless Fume Hood,
[00:02:59.465]so that's designed to capture fumes and odors
[00:03:02.212]from a certain class of chemicals.
[00:03:04.156]Next is a ventilated enclosure for histopathological work.
[00:03:08.143]And the last one is a Class I Biological Safety Cabinet.
[00:03:11.117]All those devices look pretty much the same, or similar,
[00:03:14.976]but they do very different things.
[00:03:16.349]So we have to make sure what we're working in
[00:03:19.420]and have an understanding of what it can do and cannot do.
[00:03:21.848]So the really common enclosures,
[00:03:24.208]probably the big three you're gonna see,
[00:03:25.723]first of all is a Chemical Fume Hood,
[00:03:27.621]pretty obvious, pretty ubiquitous.
[00:03:30.559]Its purpose in life is to capture,
[00:03:31.877]contain, and remove chemical odors.
[00:03:35.333]So it's going to capture them,
[00:03:37.226]not let them breach out of the front of the fume hood,
[00:03:39.765]and then get them up on the roof
[00:03:41.195]and hopefully eject them from the roof of the building,
[00:03:43.447]so they're not re-entrained back into
[00:03:45.159]the building's ventilation system.
[00:03:46.681]So that's it's purpose in life.
[00:03:48.562]If you look at the hazards
[00:03:50.028]that you may encounter in the lab,
[00:03:51.541]you really, you obviously don't want to use
[00:03:53.195]a standard fume hood with biohazards with or radioisotopes.
[00:03:57.406]Obviously it's good with flames, or fire,
[00:03:59.884]in certain conditions,
[00:04:02.370]and obviously it's meant to work with toxic chemicals.
[00:04:06.197]The next one is a Clean Bench,
[00:04:09.501]and a Clean Bench?s purpose in life
[00:04:10.957]is to protect the product,
[00:04:12.073]and that's its only real purpose in life,
[00:04:13.543]is to protect the product from particular contamination.
[00:04:16.352]So what we have (get my pointer),
[00:04:20.005]what we have is room air flows into the top,
[00:04:22.377]a blower pressurizes that air,
[00:04:23.707]sends it through a HEPA filter
[00:04:25.168]and then that clean, particulate-free air
[00:04:26.753]blows either vertically or horizontally
[00:04:28.473]through a work area.
[00:04:30.034]So it's great for protecting the product.
[00:04:32.927]If you look at the hazards, you really don't want to use
[00:04:35.447]these with anything that's hazardous.
[00:04:36.788]Why?
[00:04:38.488]Because it's blowing back in your face
[00:04:40.488]And if you ever see a Clean Bench
[00:04:41.621]with any of these labels on it,
[00:04:43.482]Scott, run. (chuckles)
[00:04:45.705]Okay, do not walk, run away from this device,
[00:04:48.226]because something bad is going to happen.
[00:04:51.786]Last we'll talk about Biological Safety Cabinets,
[00:04:54.307]and they are different, very different
[00:04:55.742]from Chemical Fume Hoods.
[00:04:56.819]They are designed for biohazardous aerosols,
[00:04:59.283]and their purpose is to capture and contain them
[00:05:01.581]within the cabinet.
[00:05:02.472]So we're not gonna pump this up on the exhaust system
[00:05:04.844]and get it up out of the roof and hope it disperses.
[00:05:06.918]We're gonna contain the biohazard within the device itself.
[00:05:10.788]And again, if we look at our hazards,
[00:05:12.193]for most Biological Safety Cabinets,
[00:05:14.371]we really don't want to work with them with radioisotopes.
[00:05:16.960]We don't want to work with toxic chemicals
[00:05:18.823]if they're exhausting back into the room,
[00:05:20.822]if they're volatile toxic chemicals,
[00:05:22.384]that's gonna come back into the room.
[00:05:24.612]They are obviously intended for biohazards.
[00:05:28.679]So, what's the definition of a safety cabinet?
[00:05:31.337]It is a primary containment barrier.
[00:05:32.807]It's the first line of defense really
[00:05:34.219]to protect the user against biohazardous aerosols.
[00:05:38.158]At the very minimum it should protect the
[00:05:39.801]operator and the immediate laboratory environment.
[00:05:42.682]Now, depending on the classification of the cabinet
[00:05:44.707]or it's class, it may also protect the product
[00:05:47.234]or what you're working on inside.
[00:05:48.802]So Class I devices do not protect the product
[00:05:51.679]but Class II devices do.
[00:05:53.809]So you have to understand that going into it.
[00:05:55.204]Not all biosafety cabinets are the same.
[00:05:58.170]And I kind of spare you all the long-winded
[00:06:00.371]definition of all these different classes of cabinets
[00:06:02.367]but those can be found in CDC's BMBL which is online,
[00:06:07.164]or NSF International.
[00:06:08.835]So these two organizations really kind of
[00:06:10.719]define what a cabinet is and
[00:06:12.179]if there's changes in the definition
[00:06:14.242]Some cabinets have become obsolete.
[00:06:16.058]Some new types have been created.
[00:06:17.870]It's really these two organizations,
[00:06:19.493]the CDC and NSF International
[00:06:21.354]that kinda control the definition of cabinets
[00:06:23.657]here in the United States.
[00:06:26.591]So who is NSF?
[00:06:27.523]Well if you're working on a Class II Cabinet,
[00:06:29.406]hopefully you're seeing this little blue mark
[00:06:31.137]somewhere on the cabinet.
[00:06:32.871]NSF is a not-for-profit, independent test agency,
[00:06:36.845]not governmental.
[00:06:38.091]So they're very much like Underwriter's Laboratory.
[00:06:40.892]Underwriter's Laboratory primarily works
[00:06:42.434]with electrical devices,
[00:06:44.579]NSF is primarily interested in public health.
[00:06:47.240]So you guys have probably seen this little blue mark.
[00:06:49.347]If you go out to a restaurant,
[00:06:50.481]you look on the soda machine that distributes the soda,
[00:06:53.939]or you look at a refrigerator
[00:06:55.374]or any other kitchen appliances,
[00:06:56.841]a lot of times you'll see that NSF mark.
[00:06:59.321]What does that mean?
[00:07:00.333]Well, NSF develops consensus standards
[00:07:03.504]so back in 1976, the user community said to NSF,
[00:07:07.822]?We would like to have a standard
[00:07:09.194]for construction and testing of biological safety cabinets?
[00:07:11.751]and NSF said, ?okay, we'll develop one.?
[00:07:14.024]So when they got together,
[00:07:15.287]members of industry like us,
[00:07:16.575]like Labconco, and Baker, and Newaire,
[00:07:18.693]they got together some people
[00:07:20.239]from the federal government
[00:07:21.541]who were interested in biosafety.
[00:07:22.997]They got together some users
[00:07:24.093]and they said let's develop a standard,
[00:07:25.859]and that standard defines how the cabinet
[00:07:27.857]has to be constructed and designed,
[00:07:29.681]and also its performance.
[00:07:31.046]So they say, these are the tests -
[00:07:32.924]airflow tests, containment tests,
[00:07:35.575]construction tests, that the cabinet must pass
[00:07:37.984]to bear the mark.
[00:07:39.722]Once we pass all these tests,
[00:07:41.743]NSF allows me to list a cabinet for a period of five years,
[00:07:45.706]but once a year they'll come in unannounced
[00:07:48.777]and audit the plant.
[00:07:50.307]They'll audit the blueprints,
[00:07:51.667]they'll audit the test records, and
[00:07:53.145]they'll audit whatever they feel like auditing
[00:07:55.202]to make sure the cabinet we're manufacturing
[00:07:57.193]matches the one that they got and tested.
[00:08:00.353]So if you're working a Class II device,
[00:08:02.415]I would really look for this mark.
[00:08:04.006]And if that mark is not on the cabinet
[00:08:05.359]I'd start asking why, alright?
[00:08:07.733]There are some specialty cabinets that cannot bear
[00:08:09.743]the NSF mark because they couldn't be tested,
[00:08:12.632]because of their construction.
[00:08:15.834]So how do you classify, what
[00:08:16.951]we're talking about, Class II cabinets?
[00:08:18.437]Class II cabinets are probably 95% of the enclosures
[00:08:21.393]you're gonna see as far as biological safety cabinets,
[00:08:24.034]and there's a hierarchy here.
[00:08:25.639]The first type is the Type A,
[00:08:27.706]and there's A1, A2.
[00:08:29.113]A2 is probably 90% of the cabinets out in the labs today.
[00:08:33.637]That's what we're probably gonna be concentrating on
[00:08:35.091]a little bit here today, and talk about that.
[00:08:38.591]There's another type, Type B,
[00:08:40.034]and we'll explain the difference between these two.
[00:08:40.928]But there's a B1 and B2
[00:08:42.608]and they finally added a new type here
[00:08:44.112]in the last couple years called a Type C.
[00:08:46.506]So it really gets confusing what does all this mean?
[00:08:49.121]The prices are different.
[00:08:50.484]The requirements are different.
[00:08:53.194]Why do I choose?
[00:08:54.096]Well, As are really designed for
[00:08:55.951]routine microbiological work
[00:08:57.779]and I should have added the word in
[00:08:59.295]volatile toxic chemicals.
[00:09:01.409]Okay, so if I'm not working with volatile toxic chemicals
[00:09:03.565]I'm doing routine biological work,
[00:09:05.133]a Type A cabinet is perfectly suitable
[00:09:07.147]for the type of work I want to do.
[00:09:09.410]Type B was developed to handle volatile toxic chemicals.
[00:09:13.625]The way a B cabinet works,
[00:09:15.001]some or all of its air is exhausted directly
[00:09:17.896]through an exhaust HEPA filter
[00:09:19.337]and not recirculated within the cabinet.
[00:09:21.478]So these guys are really intended
[00:09:23.035]for volatile toxic chemicals.
[00:09:25.887]And a C, well it's kinda a hybrid of these two types.
[00:09:29.396]So it can be, it has the insulation flexibility of an A
[00:09:32.495]but it will also remove toxic chemicals and fumes
[00:09:35.022]like a B will.
[00:09:37.065]The point of all this is,
[00:09:38.603]is A, B, or C, all contain biohazardous aerosols
[00:09:42.411]as the same, they're all tested the same,
[00:09:44.687]the air flow patterns are typically the same.
[00:09:46.978]Really, the main reason you go between A or B or C
[00:09:50.035]is not biohazards, it's chemical hazards.
[00:09:54.016]So unfortunately a lot of people think, well,
[00:09:56.340]B and C are further down the alphabet, and they cost more,
[00:09:59.031]so it's gotta be a better safety cabinet.
[00:10:01.121]No, it just has a different purpose in life.
[00:10:05.148]So, how do these things work?
[00:10:07.379]Well, there's one with the panel off, inside,
[00:10:10.183]and it's really big.
[00:10:11.384]It's just really a large air filter system,
[00:10:13.543]so I've got two HEPA filters,
[00:10:15.852]and we'll talk about what HEPA means in a second.
[00:10:19.434]I've got a motor blower, here in the blue,
[00:10:21.749]that's gonna force the air through this cabinet,
[00:10:25.921]and then we're gonna have some kind of electronics
[00:10:27.867]to tell that blower to speed up for slow down
[00:10:29.621]to maintain proper air flows hopefully.
[00:10:32.761]And then lastly, we have duct work
[00:10:33.994]so we have a grill at the front here,
[00:10:35.517]we have grills at the back, grills at the side,
[00:10:37.547]and internal duct work.
[00:10:38.656]That's gonna manage how the air flows through this device
[00:10:41.456]and how it protect the operator.
[00:10:46.250]So HEPA filters, HEPA stands for
[00:10:48.086]high efficiency particulate air,
[00:10:50.042]and that tells you what the filter is.
[00:10:51.399]It's a particulate air filter,
[00:10:53.899]it really only,
[00:10:55.090](whoops my battery's dying),
[00:10:55.923]It really only works with particulates.
[00:10:58.090]Gases, vapors, will flow through the filter, alright.
[00:11:02.022]So, if it's a particulate, if it's a droplet,
[00:11:04.196]the filter will catch it. If not it tends to go through.
[00:11:11.447]So I've got the filter, how does all this work?
[00:11:13.541]Well here's a cross section of the cabinet.
[00:11:15.265]So the blower is gonna pull room air in at the front,
[00:11:17.800]at the grill here.
[00:11:18.824]That's gonna provide an air curtain to protect the operator.
[00:11:21.536]So that inflow of air is gonna protect you
[00:11:23.119]from anything that would breach
[00:11:24.896]out of the work area of the cabinet.
[00:11:27.451]That air flows under the work surface up the back
[00:11:29.993]and the blower pressurizes that air,
[00:11:32.032]sends some through a supply filter
[00:11:33.994]and some through an exhaust.
[00:11:36.036]So the air that flows down through here
[00:11:38.147]is hopefully laminar flow,
[00:11:39.571]so you see I've got all these arrows
[00:11:41.142]moving in the same direction at about the same speed.
[00:11:44.262]Why is that critical?
[00:11:45.507]Well if I generate an aerosol in laminar flow,
[00:11:48.166]it's usually conducted right to a grill.
[00:11:50.048]If I have turbulent air flow in here it could waft around,
[00:11:52.614]float, here, there, and everywhere
[00:11:54.402]and settle where you don't want it.
[00:11:55.824]So laminar flow will contain any aerosol
[00:11:57.932]and hopefully pull it to either a front
[00:12:00.083]or back grill in the work area of the cabinet.
[00:12:03.285]So that's how this thing works.
[00:12:08.038]Now, a couple concepts, and most users don't get this, is,
[00:12:11.571]we showed the air flowing down,
[00:12:14.240]there's a vertical plane in the work area
[00:12:16.754]and that's called the smoke split.
[00:12:18.466]At the smoke split, air either goes forward or backward.
[00:12:23.275]It's nice to know that because
[00:12:25.355]if I work behind a smoke split
[00:12:27.482]or store critical samples behind the smoke split,
[00:12:29.936]so I'm back here, if this is biohazardous,
[00:12:32.913]for that biohazard to get out and get to me,
[00:12:35.324]first of all it has to break this air curtain.
[00:12:38.234]Then it has to break this air curtain,
[00:12:40.526]and then it has to break this air curtain.
[00:12:43.056]So the best chance of protecting myself
[00:12:45.108]from biohazard is behind that smoke split.
[00:12:47.800]Conversely, if I have something I really wanna protect
[00:12:50.508]from external contamination, where is the safest spot?
[00:12:55.219]Behind the smoke split. Again,
[00:12:56.977]I have to breach this curtain, this one,
[00:12:59.010]and finally get back into here
[00:13:00.825]to get to what it is I'm trying to protect.
[00:13:03.165]Now how do I find the smoke split?
[00:13:05.307]Well certifiers, when they certify the cabinet,
[00:13:07.140]usually have chemical smoke sticks
[00:13:08.938]and they can show you that.
[00:13:10.301]If you don't have that,
[00:13:11.717]you can get a beaker of alcohol,
[00:13:13.675]isopropyl or ethanol, it doesn't matter,
[00:13:15.572]and a piece of dry ice.
[00:13:17.506]Drop it into the beaker
[00:13:18.867]and for about 10 minutes you get cold smoke
[00:13:21.131]off that ethanol,
[00:13:22.387]and you can actually move this
[00:13:23.617]in the front center of the work area,
[00:13:25.351]backward and forward until you find a smoke split.
[00:13:27.729]When you found it,
[00:13:28.699]get some tape and mark the side wall of the cabinet
[00:13:31.061]and there's your smoke split, okay?
[00:13:33.957]Pretty easy, now, for some operators,
[00:13:36.115]particularly shorter operators,
[00:13:37.601]it's gonna be hard to work behind the smoke split,
[00:13:39.785]but it's still valuable information.
[00:13:41.256]It's still good to know that's where I wanna store stuff
[00:13:44.687]to keep it safest, and to keep me safe.
[00:13:49.827]So, the really critical thing other than the smoke split
[00:13:52.810]is the importance of that front grill.
[00:13:55.479]And we see a lot of users will block that
[00:13:57.123]and there's a vacuum here, air's trying flow in.
[00:13:59.099]It's really great, I can take my instructions
[00:14:01.096]and put 'em down on the grill,
[00:14:02.537]and there they are, the vacuum holds 'em down.
[00:14:04.249]I can go to town,
[00:14:05.187]it's gonna hold 'em there for me,
[00:14:06.390]I don't have to look all over the place.
[00:14:07.641]If I block that grill,
[00:14:09.349]the air still wants to flow into the cabinet.
[00:14:11.702]It's gonna go over that obstruction,
[00:14:13.943]across the work area and then get to this back grill.
[00:14:17.436]So I've just contaminated everything
[00:14:19.259]in the path of what I just blocked that grill.
[00:14:22.737]I really just basically wiped out the product protection
[00:14:26.023]in that region of the cabinet.
[00:14:27.715]And now room air is gonna be drawing over that
[00:14:29.484]and I've pretty much contaminated
[00:14:30.620]whatever it is I was working on.
[00:14:32.609]So, two things in this, if you can get the smoke split,
[00:14:35.604]and don't block the grill.
[00:14:38.015]Those are probably the biggest problems
[00:14:39.662]in having contamination inside of a cabinet.
[00:14:41.860]And we'll get those calls,
[00:14:43.809]?Oh, the cabinet's contaminated my tissue culture.?
[00:14:46.644]This is a big stainless steel box with air filters in it,
[00:14:49.483]how could it contaminate anything?
[00:14:51.722]Okay, things don't grow inside this, alright.
[00:14:54.859]Usually the contamination can be traced back to
[00:14:57.325]either contaminated product to begin with,
[00:14:59.309]or bad technique.
[00:15:02.903]So, now, I lecture on working in clean rooms.
[00:15:04.233]I lecture on working in clean rooms
[00:15:07.327]to beginners in clean room technology.
[00:15:10.476]And if you think about it,
[00:15:11.417]the HEPA filters in a clean room are exactly the same
[00:15:13.334]as the HEPA filters in a Biological Safety Cabinet.
[00:15:15.677]So in essence, what I have inside that cabinet
[00:15:17.862]is a mini clean room.
[00:15:19.293]So the rules and physics still apply.
[00:15:21.834]And one of the cardinal rules -
[00:15:23.290]I tell users of the clean room is don't move fast,
[00:15:26.111]because if you move fast in a clean room
[00:15:27.540]you're gonna create turbulence by your body
[00:15:29.972]and contaminants will drag
[00:15:31.425]all the way across the room with you.
[00:15:33.263]The same thing happens in a Biological Safety Cabinet,
[00:15:35.985]with your arm, or large objects,
[00:15:37.930]if I move them quickly, laterally,
[00:15:40.007]or particularly in and out of the cabinet,
[00:15:42.500]I'm gonna drag contaminants with me.
[00:15:44.827]So movements should be really kind of slow and deliberate
[00:15:48.355]when you're moving inside the cabinet,
[00:15:50.010]especially large objects.
[00:15:52.543]The second thing we tell them is
[00:15:53.814]think about how you're working.
[00:15:55.272]I have a field of sterile air flowing down,
[00:15:57.886]if I put something contaminated on top of something
[00:16:00.196]that's sterile, what did I just do?
[00:16:04.490]I just contaminated whatever it was
[00:16:05.951]that I was trying to protect.
[00:16:07.062]So you see this guy has got his hand above the beaker,
[00:16:09.462]or flask, that's the worst thing you can do.
[00:16:12.074]Because you're causing turbulence off of your arm,
[00:16:14.124]you can see the air is eddying here under his arm.
[00:16:16.446]So I've got all kinds of turbulence here,
[00:16:17.939]but it's also washing whatever's on his lab coat
[00:16:20.005]and on his gloves onto whatever it is
[00:16:22.384]he's trying to protect.
[00:16:23.659]So you don't wanna hover above, directly above,
[00:16:26.717]what you're trying to protect.
[00:16:30.330]And then lastly if you look at pictures in Cleanrooms,
[00:16:32.439]you never see shelves, why?
[00:16:34.550]Well because shelves are large obstructions
[00:16:36.440]That cause turbulence.
[00:16:37.802]So if I've got a shelf here in the clean room,
[00:16:39.433]I've pretty much knocked out
[00:16:40.599]any kind of laminar flow in that area.
[00:16:42.914]There's gonna be all kinds of turbulence,
[00:16:44.414]same with large objects, okay.
[00:16:47.145]We wanna do the same thing in safety cabinets.
[00:16:49.081]We really don't wanna have a lot of large, big,
[00:16:51.471]objects in that cabinet, particularly near the grills.
[00:16:53.949]It's gonna play the devil with the airflow in that cabinet,
[00:16:56.825]and that's gonna cause cross contamination issues.
[00:16:59.774]So, those clean room rules
[00:17:01.352]really actually work in the cabinet very nicely.
[00:17:07.118]And we'll talk a little bit about ergonomics,
[00:17:08.731]and this is usually where things go really bad.
[00:17:11.709]Most users don't practice good ergonomics.
[00:17:14.427]Now what we've found,
[00:17:16.235]in our ergonomics studies in the cabinet is,
[00:17:18.318]if the work surface height,
[00:17:19.955]and I'm going off the work surface here,
[00:17:21.567]is 32 inches off the floor,
[00:17:24.469]and you have a proper footrest,
[00:17:25.759]and I mean a nice wide one, that's adjustable,
[00:17:28.571]that cabinet with proper seating can accommodate
[00:17:30.484]from the fifth percentile female,
[00:17:32.954]which is five feet tall,
[00:17:34.382]to the 95th percentile male which is six foot two.
[00:17:38.295]So I can accommodate most of my users
[00:17:40.207]with a standard cabinet
[00:17:41.418]provided I have about that height
[00:17:42.932]and the proper equipment to support 'em.
[00:17:45.115]So they really have to have a footrest
[00:17:46.691]for those shorter operators.
[00:17:48.052]They don't wanna be dangling their legs.
[00:17:51.844]And these, quite frankly, you can get these
[00:17:53.737]at the office supply stores.
[00:17:55.537]The nice wide ones that tilt and are adjustable,
[00:17:58.153]those work very nicely.
[00:17:59.415]You wanna make sure though it's wide enough
[00:18:00.845]that I can get my feet on there
[00:18:01.872]and I can actually move them around a bit so I can
[00:18:03.337]change my posture and position
[00:18:05.369]as I'm working in that cabinet.
[00:18:08.134]Don't let your feet dangle,
[00:18:09.780]that's really gonna hurt you
[00:18:10.888]after a few hours of working in that cabinet.
[00:18:13.792]And then big thing is, this is not a storage area,
[00:18:17.120]you shouldn't have boxes
[00:18:18.509]or anything underneath that cabinet.
[00:18:21.344]If you look at this drawing,
[00:18:22.393]that person in essence is almost pulled
[00:18:23.613]and they can go up a little closer actually,
[00:18:25.731]pretty much to the entire depth of that cabinet.
[00:18:28.663]As soon as I start seeing boxes,
[00:18:32.516]you know, equipment, underneath the cabinet,
[00:18:34.482]I know the operator can't get in there and work properly.
[00:18:36.597]It's gonna be bad for them.
[00:18:41.145]Again, wanna support your legs,
[00:18:43.115]so we have a decent chair.
[00:18:44.678]We don't wanna have pressure points under the knee.
[00:18:49.211]They, again, have to be able to stretch,
[00:18:50.787]change your posture, change your position.
[00:18:52.414]That's good for your legs.
[00:18:53.460]That's also good for your arms, your shoulders,
[00:18:55.422]and particularly your eyes.
[00:18:56.970]You've gotta be able to move to several different
[00:18:58.917]postures or positions.
[00:19:01.212]Again, you should be able to pull up all the way.
[00:19:03.172]There should not be anything underneath there.
[00:19:04.904]I know it's a temptation.
[00:19:06.664]But the only other worse thing you can do
[00:19:08.025]is put something on top of the cabinet
[00:19:09.236]and block the exhaust air flow.
[00:19:10.743]I see people with boxes up on top of there, so,
[00:19:13.107]you really shouldn't be storing anything around the cabinet.
[00:19:16.986]And, again, take breaks.
[00:19:18.834]You're not gonna work, you really probably
[00:19:20.755]are not gonna work very well in the cabinet if you say,
[00:19:22.569]?I'm gonna go work four or six hours straight in this thing.?
[00:19:25.077]The last couple hours are probably gonna be pretty brutal.
[00:19:27.658]So you need breaks.
[00:19:29.438]Alright, again, a good chair,
[00:19:31.538]looking for something reasonably ergonomic.
[00:19:33.785]Weight should be distributed evenly,
[00:19:35.748]and use the backrest, alright. Again,
[00:19:38.972]match the back rest to your back.
[00:19:40.518]Most people don't do this,
[00:19:41.976]they just quick get up and sit at the machine.
[00:19:44.882]Whoops.
[00:19:48.059]Alright, again, make sure you're comfortable
[00:19:50.267]with what you're doing.
[00:19:52.093]Make your adjustments,
[00:19:53.376]be able to shift several times during your work,
[00:19:56.461]play with the adjustments in the chair.
[00:19:58.389]You should have several different positions of that chair
[00:20:00.465]where you feel good in working in that cabinet.
[00:20:02.356]And if you do have multiple operators at different heights
[00:20:04.596]you may wanna have a chair for each one of those operators
[00:20:07.371]because they're gonna be fiddling with that chair.
[00:20:09.491]And a lot of times people don't readjust it,
[00:20:11.874]they just well, I'll just suffer through this
[00:20:13.602]for an hour or two, and they're miserable by the end of it.
[00:20:16.115]So, you might wanna look at having dedicated chairs
[00:20:18.236]if your operators are, you know, vastly different heights.
[00:20:24.476]Ergonomics, again, a lot of people don't think
[00:20:26.278]about their hands and wrists.
[00:20:27.846]Yes, this is the working of a mouse,
[00:20:29.123]but the same things apply here.
[00:20:30.972]Now this is a nice shot, here's a person,
[00:20:32.993]if you look at their wrists, their wrists are aligned,
[00:20:35.559]they're not cranked, they're not angled,
[00:20:38.546]they're nice, straight, natural, neutral position.
[00:20:41.947]Alright, don't anchor your wrists
[00:20:44.198]Alright, don't anchor your wrists
[00:20:45.756]like they have here with the mouse.
[00:20:47.246]Don't anchor the wrist on the grill of the cabinet,
[00:20:49.646]the front, that's gonna cause you some grief.
[00:20:52.229]Again, keep everything straight if you can.
[00:20:55.738]Move the work around
[00:20:56.713]so that you don't have to twist your hands
[00:20:58.369]or arms or wrists around to get to it.
[00:21:01.069]Shoulders. Again, you wanna mainly natural position,
[00:21:04.412]slightly relaxed, if you're working in the cabinet.
[00:21:07.108]You may have to have, for taller operators,
[00:21:08.921]To either raise the height of the cabinet,
[00:21:11.395]and in some cases, if you have a really tall operator
[00:21:13.610]over six foot two,
[00:21:14.513]you may have to look at having a 10-inch sash opening
[00:21:16.468]for that cabinet so they can work comfortably
[00:21:18.419]in the cabinet.
[00:21:21.966]Again, relax.
[00:21:23.718]hThe operators are gonna have to do this,
[00:21:25.509]move around, relax, take deep breaths.
[00:21:30.568]Vision. Again, if you're doing detailed work here,
[00:21:34.174]it's going to be very strenuous on your eyes.
[00:21:40.497]Take breaks, that's what we really say,
[00:21:42.385]and again, periodically look away
[00:21:43.843]And have a distant focal point.
[00:21:44.983]Have a poster, art work, something else
[00:21:47.088]at a different focal point that you can
[00:21:49.357]take a look away at that for several minutes
[00:21:52.126]to give your eyes a break.
[00:21:54.835]Again, make this part of the stretch, break, rest routine.
[00:21:58.275]Set an alarm, to do that maybe every 15 minutes,
[00:22:00.716]or every half hour so that you know
[00:22:02.501]it's time to take a break for a couple minutes.
[00:22:03.961]While you're sittin' there, stretch, and reposition.
[00:22:08.125]And, obviously, keep the glass clean.
[00:22:10.620]A lot of these, we go into the labs
[00:22:11.922]and the glass is absolutely filthy.
[00:22:13.418]You have to look through that for hours on end,
[00:22:15.827]the glass should at least be clean.
[00:22:20.926]Alright, obviously you wanna have good eyesight.
[00:22:23.677]Make sure it's corrected.
[00:22:25.192]And my wife has this problem,
[00:22:26.665]she wears bifocals now.
[00:22:28.658]If she's working in the cabinet,
[00:22:29.874]she has a pair of monofocal glasses,
[00:22:31.936]that are set for like doing computer work,
[00:22:33.963]so she's fixed at several feet distance.
[00:22:36.610]That's what she uses when she works in the cabinet,
[00:22:38.455]rather than trying to use bifocals
[00:22:39.784]cause you're constantly doing this,
[00:22:41.482]trying to find the right focal length.
[00:22:43.765]So, monofocal glasses
[00:22:45.454]would probably be more comfortable than bi or trifocals.
[00:22:49.596]Again, resting the eyes, remember to blink.
[00:22:52.319]Most people get intense about this,
[00:22:54.124]their breath shortens, they get focused,
[00:22:56.656]their eyes don't blink, they get tensed up,
[00:22:59.179]and again, after a while you get to be very uncomfortable.
[00:23:01.018]So remember to take the breaks.
[00:23:05.837]Glare. You may have glare issues in the lab,
[00:23:08.700]and really, the point of that is,
[00:23:12.109]don't change your body position to fix the glare.
[00:23:15.043]Actually fix the glare.
[00:23:17.108]So, if you can turn off a light,
[00:23:18.376]or block a light or obstruct the light,
[00:23:20.422]or shield the light,
[00:23:21.723]that's gonna be a lot better for you in the long term,
[00:23:23.847]than you cranking around to some unnatural position
[00:23:25.958]so that the glare is not there on the sash screen.
[00:23:34.157]Alright, so how do we work in a safety cabinet?
[00:23:35.879]And this is just kinda a generic procedure or routine.
[00:23:39.949]Most people should go through them one way or another.
[00:23:42.612]So, the most important thing is,
[00:23:45.361]plan your work and work your plan.
[00:23:48.113]Know what you're gonna do in that cabinet
[00:23:50.178]for however long you're gonna be in that cabinet.
[00:23:51.927]How long is it gonna take
[00:23:53.104]and then what do I need to do that job?
[00:23:55.761]So don't drag half the lab over to the cabinet
[00:23:58.014]and try and put it in there because well,
[00:23:59.737]I may need this, but I'm not sure.
[00:24:01.817]Plan it out, it's gonna go a lot smoother.
[00:24:05.398]If you can, hopefully the cabinet is in an area of the lab
[00:24:07.757]where there's not a lot of traffic.
[00:24:10.232]If there is, you should alert your colleagues to
[00:24:12.453]not walk back and forth in front of the cabinet
[00:24:14.289]because they're gonna disrupt the airflow in and out
[00:24:16.386]and they're gonna disrupt me.
[00:24:17.916]So, if you're in a traffic area,
[00:24:19.817]try to arrange for signage or warning as a courtesy
[00:24:23.004]to not have a lot of cross traffic
[00:24:24.547]in front of that device while you're working in it.
[00:24:28.225]Alright, yes, you should have
[00:24:29.543]some kind of a disinfectant and spill clean-up kit.
[00:24:31.981]You may have a large or gross spill kit
[00:24:33.678]next to the cabinet or near the cabinet,
[00:24:35.659]and then only say a spray bottle
[00:24:37.526]and the paper toweling you need for the small jobs
[00:24:39.667]inside the cabinet. But it should be,
[00:24:42.267]that you have something in the cabinet
[00:24:43.416]so that if there is a breach,
[00:24:44.413]you shouldn't have to be reaching in and out of the cabinet,
[00:24:46.741]because what's gonna happen,
[00:24:47.656]well, I'm dragging whatever it is I just aerosolized
[00:24:49.305]out of the cabinet trying to clean up the mess.
[00:24:51.918]So have some disinfectant
[00:24:53.781]and clean up materials ready to go.
[00:24:57.051]Now, does campus have a rule about UV lights?
[00:25:03.095]Okay, what?
[00:25:04.308](audience member mumbles)
[00:25:05.317]Don't recommend them, okay.
[00:25:06.705]Some people like using UV lights.
[00:25:08.485]Obviously turn off the light,
[00:25:10.254]most cabinets have an interlock
[00:25:11.560]where you can't open the sash
[00:25:13.183]or the UV light will automatically turn off.
[00:25:15.857]But obviously turn the thing off.
[00:25:18.355]The sash has to be at its proper height.
[00:25:21.022]These cabinets draw in a certain volume of air.
[00:25:23.659]The cabinet doesn't know what the sash height is,
[00:25:26.061]it just knows to alarm if it goes too high.
[00:25:28.232]If I open that sash up really high,
[00:25:30.771]what happens to the inflow velocity?
[00:25:34.820]It goes down, it's a bigger open area,
[00:25:36.779]so that inflow of oxygen is gonna drop,
[00:25:38.601]my protection just dropped,
[00:25:40.506]So that sash has to be set at its proper height.
[00:25:44.298]and we'll get those calls occasionally,
[00:25:45.498]:How do I kill that sash alarm??
[00:25:47.770]You don't wanna kill that alarm.
[00:25:49.070]It's telling you something's wrong,
[00:25:50.930]don't use me this way,
[00:25:53.239]put the sash in it's proper position,
[00:25:55.451]let the cabinet run.
[00:25:57.131]Obviously check all the grills for obstructions.
[00:25:58.996]We talked about what happens if that front grill is blocked.
[00:26:01.064]Make sure it's clear.
[00:26:02.653]After five minutes, the air in that cabinet's
[00:26:04.398]gonna be as clean as it's ever gonna be,
[00:26:06.584]actually, within about two or three minutes,
[00:26:07.808]it's as clean as it's ever gonna be.
[00:26:09.346]It doesn't have to run and warm up for half an hour.
[00:26:11.947]All you're doing there is just basically wasting filter,
[00:26:14.841]alright, refiltering air that's already clean.
[00:26:17.172]So, within five minutes it's in great shape.
[00:26:21.131]Big thing - think about this,
[00:26:23.020]I'm gonna stick part of my body inside that cabinet,
[00:26:26.457]and I'd feel the sterile air to work on something.
[00:26:29.337]I should wash my arms and hands down first,
[00:26:31.560]and then I should glove up and I should have
[00:26:33.279]some kind of protection over my arms. Why?
[00:26:36.307]Because when I stick my arms inside that air curtain,
[00:26:40.008]what's happening to all the stuff on my arm?
[00:26:41.879]All the dead skin, the bacteria, hair,
[00:26:44.613]everything else is gonna be blowing down on that field.
[00:26:48.101]So I wanna really wear a lab coat
[00:26:49.894]preferably with over-the-cuff gloves,
[00:26:52.798]and preferably with knit cuffs if I can get 'em.
[00:26:57.446]Alright, you should wipe down the cabinet
[00:26:59.518]every time you use it.
[00:27:00.638]I know people say, I use the UV light on,
[00:27:02.761]I don't have to wipe down the cabinet.
[00:27:04.369]Yes you do.
[00:27:05.494]UV will not penetrate dirt or grease or oil or protein,
[00:27:09.814]if that stuff is on the work surface.
[00:27:12.532]You can leave the UV light on till Doomsday
[00:27:14.996]and it will not disinfect that material.
[00:27:18.207]So, employ good housekeeping.
[00:27:19.596]You should always use an appropriate disinfectant.
[00:27:21.656]Most people use ethanol or isopropyl.
[00:27:24.737]Be aware though if I'm dealing with
[00:27:27.637]a contaminant that's a spore former,
[00:27:30.693]all that ethanol's gonna do is basically shine 'em up
[00:27:33.547]and move 'em around.
[00:27:34.523]It's not gonna kill 'em.
[00:27:35.899]So if you have spore formers,
[00:27:37.934]if you are working with more resistant organisms,
[00:27:40.140]you may have to use a stronger disinfectant.
[00:27:44.051]Some people will use a diluted bleach solution,
[00:27:46.900]one to ten percent bleach.
[00:27:48.408]The problem is that leaves a residue
[00:27:50.524]which is oxidative, which will oxidize the stainless steel.
[00:27:54.178]This means you're gonna have to follow that up
[00:27:56.877]with either sterile water or an alcohol solution
[00:27:58.824]to get that residue back off the stainless steel.
[00:28:01.676]So there are a number of different disinfectants you can use.
[00:28:07.910]Again, we talked about only loading what you're gonna need.
[00:28:10.438]If I'm only gonna be in that cabinet for four hours
[00:28:12.752]I don't need to have 500 pipettes in that cabinet
[00:28:15.497]unless I'm going through a lot of pipettes really quickly.
[00:28:17.974]Okay, don't overload the machine,
[00:28:19.830]it's not gonna help you.
[00:28:22.800]Again, lower the sash, make sure it's right.
[00:28:25.226]Excuse me.
[00:28:26.394]After I've loaded everything in,
[00:28:28.388]it's nice to let the cabinet run for a couple minutes.
[00:28:30.343]That's gonna purge any contaminates that I generated
[00:28:32.520]in the loading operation.
[00:28:33.801]So let it run for a few minutes before I start work.
[00:28:37.988]Keep everything at least four inches inside the sash
[00:28:40.968](cough)
[00:28:41.801]Excuse me. So I'm well within the air curtain.
[00:28:44.109]Again, we talked about being behind
[00:28:45.790]that smoke split if you can.
[00:28:47.264]Keep the really important stuff
[00:28:49.127]behind the smoke split if at all possible.
[00:28:52.272]Really important, and I don't see most users do this,
[00:28:55.521]segregate clean from contaminated
[00:28:57.498](coughs)
[00:28:58.884]Excuse me. And you want to work from clean to the operation
[00:29:01.903]to discard, and I don't want to cross back over.
[00:29:05.275]So work should move in one direction, left to right,
[00:29:07.929]or right to left, but it should flow in one direction.
[00:29:10.940]That's gonna minimize the chance for cross contamination.
[00:29:15.650]And, again, I don't want to put my arms
[00:29:17.706]in and out of that cabinet
[00:29:18.860]while I'm doing my work because again,
[00:29:20.093]I am dragging contaminants either into that work area
[00:29:22.738]or out of that work area.
[00:29:25.058]Your arms are displacing a volume of air
[00:29:26.807]when you push them inside the cabinet.
[00:29:28.089]That air has to go somewhere,
[00:29:29.408]and a lot of times it goes out.
[00:29:33.892]Alright, really wanna talk about
[00:29:36.762]avoiding a flame if at all possible.
[00:29:40.576]And there's a bunch of reasons why
[00:29:42.143]we really don't want to use an open flame.
[00:29:44.720]First is, the down flow in these cabinets,
[00:29:46.881]in the modern, typically 50 to 60 feet a minute down.
[00:29:50.761]Okay, realistically that works out to about
[00:29:53.039]a half a mile an hour, so I have a body of air moving down,
[00:29:56.004]my laminar flow is moving down at 50 to 60 feet a minute
[00:29:58.965]through that entire work area.
[00:30:00.507]I put a Bunsen burner in there. Now,
[00:30:02.493]does anybody know what the velocity of gas
[00:30:04.228]coming off a Bunsen burner is?
[00:30:06.324]It's about 350 feet a minute or higher, alright,
[00:30:09.204]and then I'm gonna set it on fire,
[00:30:11.783]so it's nice and hot so it wants to rise. Okay,
[00:30:15.256]what's that gonna do to the airflow patterns
[00:30:17.416]in that cabinet?
[00:30:19.173]Okay, it's gonna mess 'em up.
[00:30:21.372]I've got up flow against down flow
[00:30:23.275]I'm gonna have a lot of turbulence,
[00:30:25.761]and there have been studies that show
[00:30:27.383]a Bunsen burner can actually carry contaminants across
[00:30:29.529]the entire work field if it's turbulent enough.
[00:30:32.803]That's the lucky thing that happens
[00:30:34.329]if I use a Bunsen burner in a cabinet.
[00:30:36.488]Okay, I mess up the work area or possibly contaminate it.
[00:30:40.498]The glue that holds those HEPA filters together is
[00:30:43.203]polyurethane which is fairly heat resistant to a point.
[00:30:46.103]But if I get it really hot, the polyurethane melts.
[00:30:50.988]So, my HEPA filter now begins to melt inside the cabinet.
[00:30:54.620]At that point you've lost all containment, alright.
[00:30:57.561]So, about once a year, once every two years,
[00:30:59.998]we'll get the call from a customer,
[00:31:01.458]?My HEPA filter's melting.?
[00:31:03.728]Well, you've got a Bunsen burner in there don't you?
[00:31:05.954]Yeah, I do.
[00:31:07.194]And what did you do?
[00:31:08.472]Well, a little flame's good,
[00:31:10.284]so a big flame must be better.
[00:31:12.034]So, I'm gonna get one of them big old aluminum
[00:31:14.450]Bunsen burners that's about this tall.
[00:31:15.821]I'm gonna light that puppy up
[00:31:17.246]and I'm gonna get that flame going so hot
[00:31:18.911]I could melt aluminum,
[00:31:20.353]because a big flame's good.
[00:31:22.139]Well I've just melted the filter.
[00:31:24.015]In some cases, we've actually had people burn holes
[00:31:26.133]in the filter they get it that hot.
[00:31:28.287]And are you doing in the work area
[00:31:29.989]of the cabinet that has that much heat?
[00:31:31.456]But leave that for them.
[00:31:33.588]The last thing, and the most dangerous thing,
[00:31:35.494]is in A cabinets, because they recirculate their air
[00:31:37.798]Internally. If I have a Bunsen burner in that cabinet,
[00:31:39.966]Einstein lights it up, and then walks away.
[00:31:43.375]The burner goes out. What happens to
[00:31:46.009]the concentration of gas in that cabinet?
[00:31:48.591]Because it's recirculating air,
[00:31:50.065]the concentration of gas will increase
[00:31:52.131]and eventually you'll reach an explosive concentration
[00:31:54.723]and there'll be a detonation. Okay,
[00:31:57.159]you don't wanna use a Bunsen burner in a cabinet.
[00:32:00.284]What we recommend to users is go to disposable lab wear
[00:32:02.804]if possible, alright, go to a Bacti-cinerator incinerator,
[00:32:07.391]which is a small insulated heating system
[00:32:10.576]to heat up innoculation loops,
[00:32:13.392]okay, go to hot bead glass sterilizer to heat sterilized objects,
[00:32:18.436]rather than have to use a Bunsen burner,
[00:32:20.177]I've only run across one person that had to use a Bunsen burner
[00:32:22.827]because they were trying to heat seal flasks.
[00:32:24.961]If you must use a Bunsen burner,
[00:32:26.089]it better be a smart burner
[00:32:27.723]where I have to have my hand on that button
[00:32:28.896]to get it to light and if I take my hand away it goes out.
[00:32:30.677]Alright, and it better be a small flame,
[00:32:33.898]as small as it has to be, as small as possible,
[00:32:36.588]and it only has to be on while I'm doing my operation.
[00:32:40.571]Alright, again, watch out if
[00:32:42.286]I'm disrupting the airflow pattern.
[00:32:44.263]The last thing you want in there is somebody
[00:32:46.078]with an air nozzle blowing things off inside the cabinet
[00:32:48.335]because you've pretty much lost all containment.
[00:32:50.271]So watch what you're doing to the airflow patterns
[00:32:52.935]in that cabinet.
[00:32:54.009]If you do have a spill or splatter,
[00:32:55.279]keep the cabinet running, it's trying to protect you.
[00:32:57.861]Do the decontamination operation while it?s still running.
[00:33:00.541]Don't shut the cabinet off.
[00:33:03.162]Whoops, let's go back.
[00:33:05.818]This is a really nice setup.
[00:33:08.024]This is from BMBL I believe.
[00:33:09.943]If you look at this setup it's really good,
[00:33:12.144]I've got clean material here,
[00:33:14.353]my sterile media, and
[00:33:15.752]here I've got my cultures. I'm gonna do the
[00:33:17.221]inoculation operation
[00:33:18.054]and then over here I'm gonna discard either
[00:33:19.952]in a pan for the pipettes or in a biohazard bag.
[00:33:23.127]The point of that is, if you look
[00:33:24.635]at the way that work flows, it goes
[00:33:26.121]from sterile to inoculation to disposal.
[00:33:29.416]So I've minimized the risk of cross contaminating
[00:33:32.568]this media with whatever's
[00:33:34.190]happening over there in the disposal operation.
[00:33:39.343]Okay, after you're done with the work,
[00:33:41.336]back away from the machine, let it run for a couple minutes
[00:33:43.502]because if I generated any aerosols,
[00:33:45.058]I want the cabinet to purge those from the work area.
[00:33:47.340]So give it a couple minutes.
[00:33:49.117]Some people keep their hands in there for a minute or two
[00:33:51.287]after they're done working,
[00:33:53.482]and then wipe down obviously with a disinfectant of choice.
[00:33:56.375]And then periodically please lift up the work surface.
[00:34:00.021]If you're sharing that cabinet with somebody else
[00:34:02.602]you'll be surprised what's under there.
[00:34:04.272]Paper towels, small animals,
[00:34:06.196]all kinds of stuff winds up underneath that work surface.
[00:34:08.944]Trust me, information comes back from customers,
[00:34:10.930]there's been stuff like that under them.
[00:34:12.662]It's kind of frightening,
[00:34:13.897]but keep that area clean underneath there
[00:34:15.586]because it will affect the airflow pattern
[00:34:16.980]if it gets blocked.
[00:34:19.435]Obviously, clean up, get rid of the gloves.
[00:34:21.387]Remember if I'm working with something biohazardous,
[00:34:23.596]that coat and those gloves are potentially contaminated.
[00:34:26.319]Handle them appropriately.
[00:34:28.035]So that lab coat, if you're working with biohazards
[00:34:30.493]should be handled as biohazard material to be laundered.
[00:34:35.954]Shut it down, some people like to leave the cabinet running
[00:34:38.475]24 hours a day. It?s a big air filter.
[00:34:40.907]If you leave it running 24 hours a day,
[00:34:42.872]you're probably going to kill the filters
[00:34:45.033]because you're going to block them
[00:34:46.810]with contaminants from your room air.
[00:34:49.610]So I simply just shut it down at the end of the shift.
[00:34:51.917]Leave it alone, and then turn on the UV light
[00:34:53.909]if you wanna use the UV light, and then,
[00:34:57.255]do you guys certify your own,
[00:34:59.633]or do you have a contract certification?
[00:35:03.539][Male Audience Member] Contract.
[00:35:04.541]Okay, so once a year a certifier should come in.
[00:35:06.417]That certifier's gonna do a couple really critical things,
[00:35:09.387]first of all he's gonna make sure that the airflows
[00:35:11.800]are proper, for proper containment,
[00:35:13.854]and then he's gonna validate the HEPA filters
[00:35:16.158]to make sure they're still containing
[00:35:18.231]the particulate matter.
[00:35:19.235]He is an independent person,
[00:35:20.533]he is not associated with Labconco,
[00:35:22.091]or Baker or Newaire or any of those other manufacturers
[00:35:24.299]so there's no conflict of interest.
[00:35:25.871]He represents a certification house.
[00:35:28.869]And then really, when it's brand new,
[00:35:31.011]when it arrives on campus, it should be certified.
[00:35:33.334]We like to see it annually after that,
[00:35:35.387]and if the cabinet's relocated
[00:35:37.174]and I don't mean from one end of the lab to the other,
[00:35:39.122]I mean we're gonna pick this thing up on a floor jack
[00:35:41.606]put it in the freight elevator,
[00:35:42.747]drop it down a couple floors,
[00:35:44.201]move it across campus to a new building.
[00:35:46.205]Yeah, you'd better recertify that cabinet
[00:35:48.288]cause it's probably gotten bounced around in transit,
[00:35:50.361]so that's when you have to recertify.
[00:35:54.168]Alright, so what have we learned?
[00:35:57.296]Okay, what's wrong with this picture?
[00:35:59.829](audience mumbles)
[00:36:01.692]What?
[00:36:02.694]You've got a couple things going on here.
[00:36:04.680]Yeah, he's got the sash wide open,
[00:36:06.165]so what's happening to the inflow?
[00:36:07.808]The inflow's practically nothing at this point,
[00:36:10.151]so he's lost personal protection.
[00:36:12.269]What else has he done?
[00:36:13.479](audience member mumbles)
[00:36:14.653]He's got what?
[00:36:15.889]Yeah, he's got on the grill,
[00:36:17.259]he's got his arms down on the grill.
[00:36:19.268]The coat's kinda loose and floppy,
[00:36:21.568]what's the biggy?
[00:36:22.647](audience member shouts answer)
[00:36:23.879]His head's in the cabinet.
[00:36:24.883]There are two things you should not stick your head in
[00:36:27.065]while it's operating -
[00:36:28.086]one is a biological safety cabinet,
[00:36:29.086]the other's a woodchipper.
[00:36:30.509]Okay, neither one is gonna end well. Alright,
[00:36:33.316]he's got his head in there.
[00:36:34.792]So as that laminar flow is blowing down over his scalp,
[00:36:37.292]what's happening to the dandruff,
[00:36:38.860]the hair, the skin flakes, the bacteria on his head?
[00:36:42.111]It's blowing right down into
[00:36:43.814]whatever he's doing his dilution on, okay.
[00:36:46.625]Congratulations you've just contaminated that.
[00:36:49.218]Okay, and, again, if he's generating any aerosol,
[00:36:52.175]he's probably gonna get it back in the face
[00:36:54.056]because down here at the grill the airflow's really high.
[00:36:56.807]Up here it's almost nothing, okay,
[00:36:59.287]so if he generates an aerosol and gets back up into here,
[00:37:01.706]he's probably gonna be breathing it.
[00:37:03.674]So, leave the sash where it's supposed to be.
[00:37:07.324]Got two shots here, what's going on here?
[00:37:08.806]Well, let's look at this guy first.
[00:37:12.385]How's his technique look?
[00:37:17.161]Good and bad.
[00:37:18.268]Look how he's holding his sample in the pipettor.
[00:37:21.419]He's holding it properly, right?
[00:37:23.330]he's not holding it above it,
[00:37:24.867]where he's gonna contaminate it,
[00:37:26.359]he's got it angled.
[00:37:27.566]Unfortunately, he's got a short sleeve shirt on
[00:37:29.824]and no gloves, okay.
[00:37:32.972]This guy over here, same school of thought,
[00:37:36.989]except he's got a microscope in here,
[00:37:40.459]so he's got a big lump of machinery
[00:37:42.095]in the middle of that airflow.
[00:37:43.205]They've actually customized this cabinet.
[00:37:44.683]I don't know if you can see it or not,
[00:37:47.028]but they actually cut the sash in here.
[00:37:48.255]So ,I've got this indentation into the front of the cabinet.
[00:37:50.100]What's gonna happen to the airflow patterns
[00:37:52.420]in that part of the cabinet?
[00:37:53.423]They're gonna be seriously messed up, okay,
[00:37:55.378]so I've hamstrung that machine there,
[00:37:57.448]and now I'm in there working with bare arms,
[00:37:59.635]no gloves. And if you look down here,
[00:38:01.878]there are tissue culture bottles.
[00:38:04.361]Okay, he's gonna have some contamination issues.
[00:38:08.704]Alright, again, a couple related shots.
[00:38:13.747]What's going on here?
[00:38:16.777]Let's look at this one first.
[00:38:19.779](audience member mumbles)
[00:38:20.783]Pardon?
[00:38:21.789]Yeah, he's, well first of all he's got
[00:38:23.918]this blocking the grill,
[00:38:25.345]so he's knocked out any containment in that area.
[00:38:28.663]Where's his position in the cabinet?
[00:38:30.672]Pretty much working right on top of the grill.
[00:38:33.818]Okay.
[00:38:35.688]What else is going on?
[00:38:36.937]Well, there's bare skin there,
[00:38:38.567]so why'd you bother with a lab coat? Alright,
[00:38:41.416]here he's got his plates and back here looks
[00:38:45.500]like some clean stuff,
[00:38:47.134]and over here are some tubes I'm not sure
[00:38:49.802]what he's gonna do with.
[00:38:51.657]What's the workflow pattern going on in here?
[00:38:53.239]It's not linear, it's kind of circular,
[00:38:54.561]so yeah, there's gonna be probably
[00:38:56.767]some contamination issues here.
[00:38:59.235]This person over here,
[00:39:02.074]nice garb, I mean we got hair net, face mask,
[00:39:05.034]nice coat, what else is going on here?
[00:39:09.480](audience member mumbles)
[00:39:10.480]Yeah I've laid this across the grill
[00:39:12.373]so I've knocked out containment in that part of the cabinet.
[00:39:14.687]I've got some tubes that are kinda going out here.
[00:39:17.052]Really, I don't wanna have a lot of tubing
[00:39:19.005]going outside the cabinet cause that's a pattern,
[00:39:22.669]or that's a path for contaminants to get
[00:39:23.989]in and out of the cabinet.
[00:39:25.303]If I must have tubes going in and out of the cabinet
[00:39:27.531]it'd be better to support them up on the side wall
[00:39:29.610]of the cabinet, and not block the grill, cause, again,
[00:39:30.611]I'm messing with the airflow pattern
[00:39:32.360]in that part of the cabinet.
[00:39:36.118]Alright, a couple more shots.
[00:39:39.036]What have we got going on here?
[00:39:41.664]Good question, not sure.
[00:39:44.728]Near as I can figure from this,
[00:39:46.424]we've gotten something off of a fermentor,
[00:39:48.646]and we're running it through a big filter.
[00:39:51.170]So it looks like a filtration.
[00:39:52.614]Occasionally you're gonna have
[00:39:54.448]to have something big in the cabinet.
[00:39:55.675]Great, you've got this thing in the cabinet,
[00:39:57.643]you've got the filter here.
[00:39:59.268]I don't know what this has to do with micropipetters
[00:40:02.018]and two boxes of pipettes
[00:40:04.221]and a bunch of big glass bottles
[00:40:06.128]all in the work field.
[00:40:07.788]I've got this big lump of something in the cabinet.
[00:40:09.638]I really should have nothing else in the cabinet
[00:40:11.970]if I don't need it.
[00:40:13.096]I don't know how you go from filtering this
[00:40:14.991]into micropipetting operations, okay.
[00:40:18.628]Just do one thing.
[00:40:22.319]What's going on over here?
[00:40:25.132]Well, it's kinda nice,
[00:40:26.472]I've got a clipboard up here.
[00:40:28.072]So I'm not blocking the grill at least
[00:40:29.936]with the instructions.
[00:40:30.938]So, everything's up here where I can get to it
[00:40:32.125]reasonably easily.
[00:40:33.206]You've got the gloves up there, that's nice.
[00:40:34.518]This thing's kinda a Jeckel and Hyde.
[00:40:36.312]I look over here, I've got this machine
[00:40:37.540]but it's nice, I've kept it away from the side wall
[00:40:42.175]and it's away from the back,
[00:40:43.376]kinda isolated to itself,
[00:40:44.541]so it's gonna be disruptive but minimally.
[00:40:46.229]It has to be in there, that's fine.
[00:40:48.442]And then I kinda wander over here.
[00:40:50.493]What's going on over here?
[00:40:52.426]I've got half the pipettes in the lab inside that cabinet,
[00:40:54.310]and I've got some test tubes on top of them for good measure
[00:40:58.651]and I've pushed them to the inside side wall,
[00:41:01.468]and then against the back,
[00:41:02.949]and then I've got all kinds of micropipetters in there.
[00:41:04.576]What's happening to the airflow pattern
[00:41:07.134]in this part of the cabinet?
[00:41:08.944]It's just really crazy.
[00:41:11.232]You can't get to this grill where it wants to go,
[00:41:13.119]so it's gonna go that way, alright.
[00:41:14.427]So, I've, again, got the chance for cross contamination
[00:41:15.954]in here because I've got all this stuff in the cabinet.
[00:41:17.956]I'm not sure what it's there for.
[00:41:23.562]These guys, we'll start here.
[00:41:27.279](audience mumbles)
[00:41:29.437]Lot of things going on here.
[00:41:31.071]Yeah I've got the nice ergonomic steel stool.
[00:41:33.958]So, that's gonna be real comfy after a couple hours.
[00:41:36.980]I've got an extra pair of gloves
[00:41:38.666]that I put in my jeans pocket,
[00:41:40.488]so those are gonna be nice and sterile,
[00:41:43.975]I'm working bare-handed or bare-armed.
[00:41:46.461]But look at her posture.
[00:41:48.274]Her back's arched forward,
[00:41:50.745]her feet are kinda weird with lotsa stuff down here.
[00:41:53.312]I'm not close to the cabinet so I'm reaching in,
[00:41:55.689]my back's arched,
[00:41:56.931]my head is resting against the bottom of the canopy
[00:41:59.662]because I don't have the right height relationship here.
[00:42:02.428]So I'm gonna conk my head against this piece of steel
[00:42:05.086]for a couple hours, you know crap like this.
[00:42:10.032]What's she gonna be like in a hour or two?
[00:42:13.010]She's gonna be very uncomfortable, okay.
[00:42:15.603]So, lot of things there, what do we have here?
[00:42:19.921]Again, a couple things.
[00:42:21.581](audience mumbles)
[00:42:22.793]What?
[00:42:23.795]Yeah, well this guy's in a lab coat.
[00:42:25.686]These people are in street clothes.
[00:42:26.969]they've got gloves on which is nice.
[00:42:28.427]She's got a nice fuzzy sweater or something on there,
[00:42:30.298]so that's not gonna shed any particles inside the work area?
[00:42:33.642]But I got two people in a four-foot cabinet.
[00:42:36.532]There's four feet of real estate
[00:42:38.038]between these verticals here,
[00:42:39.374]so I've got two people working in four foot spaces.
[00:42:42.317]And if you look, they've both got their legs cramped
[00:42:45.825]almost 90 degrees to where their torso is.
[00:42:47.800]How are you gonna work like that for any period of time?
[00:42:53.486]You've got two feet of space for you to work in
[00:42:55.919]and for your body to be in,
[00:42:57.343]and then there's another body.
[00:42:58.860]Within a couple hours these people
[00:43:00.246]are either be best of friends or at each other's throats,
[00:43:02.507]okay, probably at each other's throats is
[00:43:05.275]the way this usually works out.
[00:43:06.909]So, that's not the right size cabinet for two people.
[00:43:09.490]Most people say you should never
[00:43:10.812]have more than one operator in a cabinet
[00:43:12.545]because of the chance for cross contamination
[00:43:14.346]between these two people.
[00:43:17.781]And last but not least,
[00:43:19.121]and I'm sorry this is a jaggy picture,
[00:43:21.063]but what do we got going on here?
[00:43:22.809]So, I've got the cabinet here and a stack of stuff behind it.
[00:43:26.023]Do you see what's down there?
[00:43:31.036]It's a propane tank, yeah,
[00:43:34.244]somebody went out to Quick Trip or whatever
[00:43:36.455]and bought a propane tank cause they
[00:43:38.177]didn't want to hook it up to house gas.
[00:43:39.817]So I've got a propane tank lying down here
[00:43:41.126]that's unsecured, connected to a rubber hose,
[00:43:43.323]that goes into the cabinet's gas jet.
[00:43:45.736]This is going to end really badly, okay.
[00:43:50.036]Excuse me.
[00:43:51.912]Alright.
[00:43:53.424]If you want further information,
[00:43:55.475]here's my contact information,
[00:43:57.802]you can go to Labconco.com.
[00:44:00.293]We have literature on different types of enclosures,
[00:44:03.811]sales literature obviously,
[00:44:05.517]you can download those as PDFs.
[00:44:09.370]Once you register, we don't spam you, send you a lot of junk,
[00:44:12.345]but you're free to download that information if you wish.
[00:44:16.719]And there's my email. Are there any questions?
[00:44:20.715]Yes?
[00:44:21.968](inaudible talking)
[00:44:27.870]Is there a minimum size of cabinet
[00:44:30.113]where two people can work simultaneously and safely?
[00:44:33.709]Excuse me.
[00:44:37.215]I've heard of some people doing it with a six foot cabinet.
[00:44:40.613]That gives each user about three feet of space.
[00:44:44.136]I wouldn't try any smaller than that.
[00:44:46.422]Excuse me.
[00:44:48.583]Any other questions?
[00:44:51.798]Okay, well thank you all for your time,
[00:44:53.790]I really appreciate it.

Using Biosafety Cabinets

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Using Biosafety Cabinets

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