Aseptic Technique
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- Опубліковано 21 вер 2024
- For more information, visit www.bio-rad.com....
This video demonstrates basic microbiological aseptic techniques used for transferring bacteria from one culture to another. Techniques include use of a Bunsen burner, transferring bacterial cultures from Petri plate to a liquid culture or from liquid to liquid culture using inoculation loops or needles.
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1:32 Liquid culture to Petri plate (loop)
2:45 Liquid to Liquid (loop)
3:45 Petri plate to Stab culture (needle)
4:40 Petri plate to Liquid (loop)
A few extra tips:
1. ALWAYS gently swirl (mix) liquid media if cells are present -- before taking from the source broth, and after adding to a new broth. You want an even suspension (cells are heavy and sink) for consistency.
2. Always work close to the flame -- the hot air prevents dust from falling and settling in your new cultures.
3. When adding colony from an agar culture using a loop to transfer into a broth, swirl vigorously to release cells from the loop.
'thanks for the addition.
I'm thrilled that I found this video, I will be doing aseptic technique on Friday in lab. This video was easy to follow and demonstrated proper technique! Thank You
just curious. what are you doing now?
Monday i will be doing my practical on aseptic techniques and i must say this was very helpful. i even feel like i have completed my practical already. thanks a lot
Got sent here by my professor! Haha
Me too😂
Wow me too😄
sameeee
Yes, we’re growing vertically!
Same
Nice info. Reminds me of my microbiology courses in the 90's.
I was really taken back when I started working in a medical micro lab. Since the sample isn't sterile to begin with, and most samples are rarely kept for more than 3 days, we don't use flame for anything! No propane, no bunsen burner, no metal loops or stabs, no reusable/auto-claved glassware.
Disposable plastic loops, plastic test tubes, pre-poured/QC'd plastic agar plates... aseptic technique is a research only technique I suppose.
That's very interesting.
Aseptic technique is used in tissue processing. Hope you're doing well today with much success!
1:30 Transfer bacteria liquid culture to petri plate
2:45 Transfer Bacteria Liquid to Liquid.
3:45 Transfer Bacteria petri plate to stab culture
4:42 Transfer Bacteria petri plate to liquid
Love this channel, makes it easier to understand the lab before going to class!
my professor said we should avoid having the handle go inside the tube as its not sterilized. In this demo he goes in with handle.
yo who's here in 2020, nah I'm just kidding
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Me
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I was always told to do with out gloves when using flame as they can “melt” we used longer equipment that you kept your hands bellow a level
Watching this a night to my Practical Pharmaceutical microbiology exams🤝
Hey @Bio-Rad Laboratories can you tell me why you're doing this in open air rather than under/in front of a Laminar Flow Hood/Biosafety cabinet, I'm a mycologist and in my experience open air transfers pretty much always contaminate.
I watched the whole thing thinking the exact same thing. This would surely cause contamination in my experience in mycology
I like microbiology but I only have one arm. Can I do all these things with hand without creating major contamination of culture such as by lying the lid of petri dishes on the table while holding something else? I can also close the lid quickly. inoculation loop I know I can stand it up in a test tube holder like the one on the video. I took microbiology lab but for the classes, errors and minor contamination weren't taken in consideration. However, I'd like to work in a lab for school to make some money and some of the duties are aseptic techniques. If I contaminate everything, I assume I'll get fired. is closing the lids quickly really important?
It's important to close the lids quick because there are air microbes (that I've seen) that could contaminate the petri dish or tube that you're trying to transfer to. The quicker/more closed the best chance you have to transfer the bacteria and only the bacteria into the dish. Edit: I'm sorry if I sound like a smarty but I'm currently taking microbiology and thats what my professor told me to do. So, hope it helps.
same but i have no arms
@@bennytoyz how do u type?
This video is very helpful! Thank you!
You are perfect. Thank you so much, it is so useful video. Good luck!
Thank you so much for this video. It will be of a great help in my laboratory class.
1:03 I cannot see the difference but for flow :( yellow is not appearing only little orange.
And he did not sterilize the last agar tube by passing it through the flame as you did for others when he transferred microbes from plate ?? is it because the tube contained another microbes in it ?
Great Job by the way I liked this great explanation.
how do you determine the sterile area that you can work in around the flame?
would the updraft from the flam draw fresh air and contiminates toward you if you work at the table level?
or it is best to hover over the flame as you work?
Not the aseptic technique I was looking for but cool.
I wonder how this compares to working under a laminar flowhood.
Thanks for making this video . supporting for laboratory class thank you so much...!
In the last part, why didn't he streak the agar with the needle obtaining a broth? He just dipped it in the broth and sterilized it and the end. I dont understand
He took some microbes from the plate first and put them in the broth.
this viddeo is very easy to follow. Thanks so much.
Thank you for very helpful tips
If some bacteria, fungus spores, and dust float then what if they float in when you open the lid? Does it not possibly contaminate the sample?
Before you start anything, make sure bench top is first sterilised with ethanol and wiped dry. You need to try to do this asap, but don't rush. Sterilise anything that's not plastic (though some of us did a quick one in school previously) with bunsen burner after opening, and sterilise again before closing the lid quickly as shown in the video. The periphery near and above the flame is taken as sterile (but please work from a safe distance carefully). Did some quick googling, and it seems like air current is brought upwards and away when there's a flame. Managed to do this during secondary school days with no contamination.
During my diploma, we will usually work within a laminar flow hood instead. I'm just getting back into studying again 8 years later, but I hope this helps. :)
@@MomoChanhs I was thinking more of the part where he opens the petri dish and smears the bacteria in it. You'd think every time you open it it would make a vacuum that sucks all surrounding air in which could suck in any spores from spore forming bacteria or molds that might happen to be in the room. Maybe I'm too picky but seems like it's about impossible to collect a sample of anything that could be proven to not be cross contaminated from another source. So in a sense I almost wander what how lab samples are proof of anything at all. I mean you can't prevent outside air and contamination from entering the dish. How do the compensate for these possible contaminations I wander? Plus the fact that some spore forming bacteria are heat resistant and not killed by cooking etc. Plus bacteria that can form endospores can lie dormant for thousands or millions of years even. Endospores can survive without nutrients. Endospores can survive without nutrients plus they are supposedly resistant to ultraviolet radiation, desiccation, high temperatures, extreme freezing, and chemical disinfectants. I've even seen where things such as alcohol pads get recalled for bacterial contamination such as this recall in this link. www.accessdata.fda.gov/scripts/cdrh/cfdocs/cfres/res.cfm?id=100988
I guess a combination of these things leave me totally unclear of how all this is suppose to mean anything as far as scientific evidence if it could all seemingly be impossible to prove its not cross contaminated?
@@naturestrail2296 As long as aseptic technique (the bunsen burner should be on throughout) is employed, chances of contamination is reduced; but I won't say it's 100%. If the flame is big enough, it should control the air flow nearby and not allow spores to get sucked in. In research projects or for company usage, usually a laminar flow hood will be available to reduce/eradicate the possibility of contamination. Laminar flow hood was only invented in 1962, so scientists had probably just settled with aseptic technique throughout history.
Meanwhile, lecturers may choose to stick with aseptic technique for efficiency in a typical classroom (I'm guessing you're a student).
Some bacteria are heat resistant, but exposure to naked flames should get them killed. Cooking temperature is usually around 100 deg. Celsius, which is insufficient. Thus autoclaving is needed. Usually a product will be required to pass QA/QC checks (and can include bacteria and endospores count) before releasing into the market. Products that do not pass the requirements are usually rejected; based on the stringency level of the production company.
In the case which alcohol pads were recalled for contamination, it might be caused by human error/mechanical faults when packing/producing the product. If the seal of a product is not properly done, the product will no longer be sterile and thus, a recall will be warranted. If an intern/a staff in the production line happened to use the wrong dilution factor (diluted too much), or even mixed up solutions, the product should be recalled. While I was in the QA/QC field previously, some suppliers would supply faulty packaging materials/make some mistakes occasionally... So it is up to the QA/QC team to check through and decide if a product is safe for release.
It is responsible of you to be picky about all these though. Keep it up! :)
I had worked with classmates/colleagues who did not care a single bit about contamination nor lab management fundamentals, putting lives of others and themselves in danger... I'd very much prefer working with someone who ask the right questions and ensure everyone is safe!
@@MomoChanhs actually my job requires lab sampling and just curious how all of it works when we send samples to a lab. To me is easier to understand and remember the sampling preparation procedures for shipment if I have a better idea of what's going on at the other side is all. Thanks for the responses. I'm sure it can help others that want to read as well. I've just always preferred to have a thorough understanding of how things are conducted.
@@MomoChanhs which I knew the burner kills on contact of the actual flame but I was kind of wandering more regarding the air flow and possibility of contaminating within the air floating around. A burner running I'm assuming would increase air flow as they pull air into the bottom to create the air/fuel mix and the heat rising with that combined would create a circular draft in the area. But I'm assuming the hood your referring to would solve that although I can't picture what the hood looks like as I've never actually seen one. Wasn't thoroughly sure how that spore forming bacteria actually works. Do the spores develop ultra fast? Or can someone easily tell the contamination resulting from spores floating in the air vs direct bacterial contamination? Is this one of the reasons for a time controlled period for the sample to be at the lab? Just kind of curious to learn more about them. So far I gather they form spores when stressed basically as a typical survival type response. About the extent of what if gathered I guess.
Sterilisation without hood.... It seems weird. I saw once in lab that my media accidently got opened outside hood and bacteria developed in it next day.
Well explained. Thanks
I cant hold the jar and the loop at one time ,what is the alternative procedure
helpful and easy video for bactrerial inoculation
it's really helpful.. Thanks before for uploading..
You have to burn a metal rod while IN A CLEANROOM to sterilize it?!? Or does a cleanroom make it so you don’t have to do all this??
Yes. A cleanroom does its best but there is always a chance for contamination. The more precautions you take, the better. Besides, rooms are not safe havens. Imagine if one day the ceiling started leaking because of storm damage? Or a bug got in and was spreading particles? A company could lose all of their current product because people trusted a room. It's simply better to be safe than sorry. You wouldn't want someone to claim your research was inaccurate because of improper technique either. The horror! (I hope I was able to give you a couple reasons why.)
Thank you so much!
Why is it needed sterilize each time needle and loop?
Thank you so much sir❤️
I find it difficult to transfer with the loop, I often break the surface of the gel when using it... The culture medium used in this video seemed very dense. In my uni, it's always very fragile, so I have to use the little glass bubble of a disposable glass Pasteur transfer pipette...
you can practice.... ur hand should be light while doing the inoculation..
I don't mean any offence, but your aseptic technique is horrible 😂 unless you're working in an already aseptic environment (flow hood, isolator, extremely clean room) your just working in the open air. The bunsen burner creates a sterile vortex of air around it, you should be working within that area. The light flaming you give the flask and test tube wouldn't be sufficient. You're better off holding the vessel with one hand, removing the lid with the same hand while as close as possible to the flame. Open the lid as little as possible, retrieve the culture and replace the lid. Keep the inoculation loop near the burner, grab your plate and place it on the bench as close to the burner as possible without touching. Open the plate, keeping hold of the lid. Streak the plate in one go and replace the lid. Flame the loop. Seal with parafilm. Done.
It's also not really the safest practice to wear latex glove around open flames. We were taught not just scrub your hand really well if doing aseptic work under a bunsen. That was both at university and in the industry.
simply
we 'll attempt or transfer the bacteria into
liquid_ to _ liquid
soild to liquid
liquid to solid
soild to semi soild
simple those all process we do in lab with the help of bunsun burner and by the inoculating loop
thk's for the video, but with the bensen berner, avoid wearing gloves .
my prof really said go watch that vid then dipped
Who’s here in 2023 😮
does anyone find out any step is missing??
i havent studied microbioloy in minor but my research topic is related to microbiology lactobacillus. plz help me recommend me gud books and youtube channels.
Interesting
Thnks for u
my bacterial hood back home is underrated :(
rad
Sent by my teacher
very nice
Best vedio
Who's watching in quarantine 😵
Great now to try with an exotic swab for the first time 🤣✌️
Sweet video
4:29 did he forget to sterilize the mouth of the tube?
Donbol Don I guess he didn't flame it coz its a plastic test tube
Thats what I would've done too. I'm not too sure either though. I would've done it just in case though
I wonder about that, too
Who is here 2023 🤔
Huhu return demo later hahaha
Hello friend watching from yolandsa
Hai kisay!
Pandy students doing virtual labs raise your hand.
mr. birney 2019 anyone?
Hi Professor Shrek
Yeh who’s here in 2023
🤚 I am seeing this in 2024
was it just me who thought it was on loop?
Still it's not ideal and need improvisation. microMan 😎
You have exposed arms while trying to be sterile? Cmon
Anyone else here for GCSE?
Bad technique with stab inoculation. Taking off the cap with the same hand that is holding the tube and fiddling with the cap is a sure way of getting contamination. Holding the cap with the little finger of the dominant hand allows for full control of the transfer and inoculation.
Not gonna lie, the organization of this video is annoying. You constantly have to work backwards with some of the information. They don't tell you to sterilize the loop with the first Transfer process, only subsequent transfer processes, same with allow the loop to cool. I am also left guessing whether to sterilize the mouth of the stab culture. I strongly feel it is important to be clear when you are working with bacteria samples and sterilization processes, but I guess this video is better than nothing
.
Wuhan needs to watch this video 😂
This is so hard to follow
kindly writedown whatever u say because its difficult to understand pronunciation
....
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