DIY Tissue Culture PART 2: Temporary Immersion
ฝัง
- เผยแพร่เมื่อ 10 มิ.ย. 2023
- Live. Laugh. Autoclave. Plants are science! Let's propagate them the scientific way.
/ plantsinjars_tc
Where I buy tissue culture supplies: www.plantcelltechnology.com/?...
Supplies List:
• The BioCoupler: bit.ly/3p2bvwj
• Murashige & Skoog: bit.ly/3pMpAOr
• Kinetin: bit.ly/41IpXaa
• BAP: bit.ly/3MYJ2zv
• IBA: bit.ly/3N0cghE
• Scale (0.01g accuracy): amzn.to/3I9fDRM
• pH Meter: amzn.to/3NkTiU6
• pH Up & Down: amzn.to/3pAM19r
• Forceps: amzn.to/42K3VFa
• mL Droppers: amzn.to/44X8cXl
Use code PLANTSINJARS for 10% off your Plant Cell Technology order.
Protocol: journals.ashs.org/hortsci/vie...
This protocol uses PPFD to measure light. To convert PPFD to lux, I use this calculator: www.waveformlighting.com/hort...
Timestamps:
1:51: What is the BioCoupler?
3:09: Why does the BioCoupler work?
4:13: Overview of the Tissue Culture Process
6:19: How to Make Liquid Tissue Culture Media
10:16: How to Sterilize a BioCoupler
12:44: Setting up the Still Air Transfer Box
14:06: Still Air Box Demo
16:49: Laminar Flow Hood Demo
17:40: Bonus Experiment
Please note that some of the links in the description are affiliate links, which means that if you click on one of the product links and make a purchase, I may receive a commission. This commission comes at no additional cost to you.
I own a licensed cannabis micro-cultivation facility. I’ve always been taking clones. Just ordered everything needed to start testing with tissue cultures. Wouldn’t love to be able to preserve genetics, and not have to maintain mother stock.
Thanks for the videos. I always assumed TC was outside of my reach being a small time craft grower.
i wish this channel existed a couple years ago, woulda saved me thousands
I cultivate Mushrooms so i got 98% of the necessary equipment. I’m going to give this a try. Thanks!
Can we cultivate Shiitake mushrooms this way at home?!
@@ImAshlynnCarter To cultivate Shiitake Mushrooms is a multi-step process starting with the creation of a liquid culture. This liquid culture is then used to inoculate a Grain spawn, which consists of sterilized grains. Once the grain spawn is fully colonized, it is transferred to bags filled with a mixture of wood dust and woodchips, (bricks) where the mycelium can continue to grow and eventually produce shiitake mushrooms.There are similarities in the techniques she is showing, but they are two different cultivation methods.
I was thinking exactly the same! Bonus, cultivating mushrooms takes an enormous amount of time for sterilizing all the grain (I have a small pressure cooker, EU problems), while plant tissue propagation appears to be less time intensive and more actual cloning work :)
Same!
I’m looking for something to make money when I retire in a few year. Realistically, is there money in this gig?
If you aren't planning to do a plant science PhD then you should! I worked as a postdoc on GM crops for 10+ years, sat through loads of presentations and 90% of the presenters were unable to explain a method like you do in these videos. A true you tube gem!
Oh and kinetin is also used in anti aging creams for anyone watching.
I love how she called everyone out for being tempted to click away. 😆
As a "stoner" I appreciate and use your info...but also I use psilocybin as medicine...oops. I find your information can parallel/apply to the propagation of both cannabis AND fungi...and maybe other stuff too🤔. I appreciate that you consider people without money to set up a viable lab...even though you know you want all the tech lol
Tip. It’s a lot easier to drop the plants in the bottom jar with the liquid media than placing them in the top jar. Also, because you have more media per jar compared to traditional tissue culture jars. I sterilize the BioCouplers for 30-40 minutes.
Thanks for the tip! The ones I set up in this video haven't been showing any signs of contamination (yet...lol, fingers crossed), but in the future I'll definitely increase the sterilization time
Such great explanations and thorough video, Laur! That African violet is looking amazing, we cannot wait to see the rest those plantlets grow and multiply in the BioCouplers!
I have never done tissue culture for plants. However, I did spend years doing microbiological research at a university for work (including bacteria, yeast, viruses, parasites, etc) and I have to say, we HAD bleach in the lab but only used it for ONE very very specific thing. Every single other decontamination process was done with autoclaving and 70% ethanol. I’m not saying there aren’t data out there that support bleach being more effective but I do personally have confidence and experience using 70% ethanol. I will also add that ethanol is more effective if allowed to evaporate completely on its own rather than be wiped. I think people who have never done sterile procedures would be inclined to “clean with it” like you’d do in a kitchen perhaps rather than just spray and ignore.
This is just my $0.02 on ethanol but every person has their own way of doing things. If you like bleach then I think you should do bleach and just remind people that ethanol is also an option. Just like you did with the plant growth regulators 😊. I like your videos and it’s exciting to see a fellow young female scientist and plant lover. The longer I watch the more I wonder where I can set up TC in my house lmao
Agreed, spent years doing animal tissue explant culture, only ever used bleach in cleaning animals housing and things like that. Tissue culture stuff has always been 70% ETOH, autoclaves and sterile filters for temp sensitive stuff.
I trust it a lot more than bleach.
I must be really old, back when I was in uni we were using phenol for decom!
Personally as a hobbiest I prefer a %10 peroxide solution over bleach, it's just as effective of an oxidizing agent as bleach and like alcohol, evaporates without residue.
Even mixes with ethanol! (I do, but I don't recommend)
I've done tons of tissue culture for aquaria, fungus propagation from spores/plugs/spawn, and growing moss in plate cultures (still tissue culture, just weird and difficult because there's not a lot of public literature). In all of the above, 90% isopropyl and 70% ethanol both worked fine for me over the years. I will admit to using a blowtorch on a steel table quite a few times though when I didn't have proper equipment, which also worked just fine lol.
Thanks for your valuable input! Do you think ethanol can be fully substituted with IPA? Where I live it's much cheaper than ethanol.
I used to have to clean an artificial insemination lab and we would always wipe it off. They would have us spray it on the towel then wipe everything down
I have 36 or so of these biocouplers. I am growing aquatic plants with the same media formula and these things are taking off! I would say that within a month and a half, the biomass of plant material increased by 20X (no joke).
Which plants are you growing?
Wow! Would love to hear more about your experience!
That's awesome. Which aquatic plants are you growing?
@@plantsinjars I am growing Alternanthera Reineckii Rosanervig and Staurogyne Repens. I keep the plants submerged in the biocoupler and only flip it and swirl them around maybe once or twice a week. Leaving them in the media makes them grow extremely fast. I purchased the explants from a local pet store, separated them into 5 couplers, then after a month subcultured them into another 10 couplers and they have since doubled in size from there (a month and a half total - and still growing).
@@zerocostcompliance5407 oh nice do a video about it
I'm a plant scientist who work in large scale plant propagation on woody plants
I'm inspired to use the Bio-couplers despite doubting whether it will vitrify (since many people said there are more challenging even use TIS for woody plants).
Thank you for sharing. Love your last segment experienment and TC Claim ❤🎉
Hi teacher! Does liquid culture need to add sucrose like solid culture?
Glad i found this channel. Great information & exactly what ive been hunting for.
This is such a great series, thank you!
Awesome!! Thank you for doing these videos! Wife and I are having a blast!
You have the best TC channel by far.
Love your channel. Not into plant cloning etc, but I love learning diverse stuff and I love the academics and scholarly nature of your videos. You’re a great resource and even my lecturers didn’t keep my attention as long as you have. Thank you!
Brilliant, thank you for another fantastic demonstration. You're really doing a good thing here, sincerely thank you.
Thank you for the detailed video. One of the best canals I have seen on this topic. Again well done. So much to learn. 👍🏻👌🏻🙏
I’m enjoying your DIY TC videos. I’m actually thinking about attempting to try it this fall.
Thank you for all the knowledge :) great setup.
You are the GOAT. I love your videos.
I really enjoyed your videos. I first learned of tissue culture in nineteen seventy six. And have always wanted to do it. Now in my retirement I plan on doing it for fun. I am a retired nursery man. My name is Michael.
As someone who as both grown and cloned marijuana I can see how someone might think cloning cuttings is basically going to get you the same result as TC in the end, but where we might get a couple hundred clones from a mother plant you could make thousands (maybe tens of thousands) of TC clones from the exact same amount of plant material. It also seems slower at first glance, but with the ability to propagate in a smaller footprint and multiply clones exponentially faster TC is definitely the way to go if someone is confident in their abilities and can get the supplies needed.
Another great video thanks, very well explained. Love your sense of humour, especially for what can be a dry subject :)
You’re absolutely adorable and super smart. Love watching your videos!! 🥰
I'm so grateful to have found your channel. I've been working in the Colorado medical industry servicing those who truly need medical access... Been working so hard to build the most amazing genetic library, and now I'm trying to get everything tissue culture for archival purposes, as well as master the meristem tissue cultures.. Your channel has so much useful knowledge and information. Thank you. Thank you thank you!!!!!
I love these videos. Thank you so much, really great information on them.
brilliant energy. very fluid delivery
Thank you I find your videos highly entertaining and educational please keep doing them ✊💯🙌 you have inspired me to try this tissue culture!! I'm an avid gardener and plant lover
Great content and you do an extraordinary job of presenting it to your audience on your TH-cam channel! Well done and keep those videos rolling out on your channel! I’m a huge fan!!!
Thank you!
Great videos. Thanks for all the structured information!
Thank you!
Am so happy too watch your videos,u put me on a hole nother level.
Awesome videos, truly inspiring! Thanks!
Wow. Found a gem in your content
You’re a natural!
not gonna lie i watched your video for my favorite plant like the others😅 your information were reliable and i was able to start my own protocol so thank you a lot you and plant cell tech made me look a tissue culture
❤❤ awesome thank you so much for sharing your knowledge❤❤❤
This is so neat! I work as an Aseptic Technician in a clean room, making veterinary pharmaceuticals! I love my job and if I could implement it at home, like in this video, that would be so awesome!
As someone with a molecular biology background, this is definitely awesome and well made! You told us that tissue culture gives you more propagations, but I would love to know if it is truly more effective than traditional water or moss propagation! In other words, what is the success rate like? I am terrible at traditional props and I usually end up with mold or rotting, even after changing the medium often and keeping humidity high, as well as keeping a sterile environment. It seems like tissue culture is more fool proof and less can go wrong. Ironically, I would be more comfortable with this than traditional props because I've done this sort of stuff in a research lab for years!
I have so much crossover lab equipment from mycology but those PGRs got me straight up shook. All just so I can clone an orchid or two.
Cool video! Thank you for such great content!
Love your work
I admire anyone chasing the Nth degree.
The clarity and simplicity with that you describe the very technical process your "hobby" makes a resounding and profound impression of a simple first impression.
You are to be cherished. Hope you heal well.
The Most important and critical thing in life overall is to simply find something to care about- and care deeply. You have done that.
You have a lucky hustband/fiance
I know you're trying to be nice, but you could have left the last part off entirely. You can admire someone's intellect and scientific contributions without making unwarranted subtle references to their perceived sexual appeal or relationship status.
Such a fun learninng experience, great to be here.
Super cool video. You did an excellent job breaking it down
This is very cool, thank you for sharing it with the world!
Having done this at uni, I have always thought about doing this at home. Still doing lots of home horticulture, maybe i should give this a go?
i never thought i would hear a cooking with jack reference on plant youtube 😭 subscribed
I love these videos! I'm in school right now for conservation biology and ecology and my dream is to have a career that would allow me to work with plants all day. I cant wait for the Q&A! I would be seriously interested in finding out where you get your lab equipment from such as the flow hood. I looked in amazon, and while I can imagine they're quite pricey anywhere, something feels wrong about ordering it from amazon lol! I cant wait for the next video!!
Thank you! The flow hood is from a company called Air Science and my hot plate is from Fisher Scientific :)
Hola from Costa Rica! My name is Eugene, and I have a couple of questions for your Q&A video next time:
1. After the actual Tissue Culture process, where do you store the jars/containers? Is it on a rack with bright lights 24/7? Could it be on a bright shade? Under the sun?
2. Just curious, how do you do the acclimation process? And how do you manage space for a number of baby plants?
i looove your videos, thanks so much for all information
I have some rare cycads that I want to try this on :) Heard about tissue culture about 15 years ago but never looked into it. I'm sure I can learn by following your directions. Thanks for making these. New sub from South Africa
Right now all my space is filled with my aquariums, but as soon as I have clean space I am definitely gonna have to join the tissue culture gang!
Neat channel. I've been doin aquatic plants and house plants/gardening for years. Havent done anything like this.
Great content I appreciate how you explain these processes. I have a comment on the bleach rinse tho, some were saying they don’t want to use bleach solution, but if you want a clean plant without viruses of infection that’s what they need. Explain to them that alcohol does not stop disease and the bleach is perfectly safe . Great content. Keep it coming. I may reach out for questions I have. 🌱💜
Nah. Use h202. bleach is made of hypochlorites -salts. Plants do not like salts.
I've always had 10% peroxide as a go-to
Never leaves a saponified residue to rinse, or any residue on anything else.
(not the 3% watered down, 30% watered down 2+1, like hair and tooth bleach concentration)
Спасибо тебе за видео, ты всё очень классной объясняешь )
I sometimes watch gardening videos so I suppose that’s why your channel showed up in my feed. Honestly, I’m not really sure what this is about. I put seeds in soil then sunlight and water make food and flowers. But you seem to enjoy what you are doing and I dig your style, so best wishes.
really great tutorials! thanks!
Thank you for this series. I am learning allot. Can you give an aproxamate total timeframe from stage 1 to stage 4 rooting?
Humor, wit, beauty and technical expertise (with a dash of plant nerdiness) !? What more can we ask for? Incredible content ⚡️ Instant sub.
I was about to relay the same sentiment, well said.😌
*tips hat*
Your funkin awesome!!! thanks for all the wonderful info!!! Its time to stick it to "Big Tissue Culture"!!!
love the videos
My family always had a pressure cooker for Pepperpot at Christmas.
I have purchased ball jars and double lids which I modified into a BioCoupler. I have just rescued my last remaining Bee Balm plant before the frost, and put it into a bio coupler along w/ growth hormones. My goal is to have a great display by next summer.
I keep losing the few good plants that do well in my 5x6 community garden plot. I hope this helps me to keep and propagate plants that I like.
Thank you for well organized information ❤
Awesome video! Wondering about last steps - dividing plants from a biocoupler, putting it in rooting media and last step of putting grown tissue culture into soil
I'm making an updated vid about that topic soon!
Wow i have to thank the yt algo for this recomandation! I wasn't aware that tissue culture is doable at home...now that i want to dive into shroomz & must say that tissue culture will be in my further experiments for now on! thank you so much for sharing this knowledge!
This biocoupler looks like some specialized loose leaf tea tumblers. They have a mesh in the coupler, you place your tea in one chamber and then the hot water in the other chamber(be careful about pressure, hot water in a closed chamber expanding air and such) you flip it to immerse the leaves in the hot water and flip it back to drain the liquid off the leaves when it's brewed to your desired strength. I wonder if those could also be used.
I absolutely love your videos you’re amazing! The memes always get me let’s go cottagecore girlies
I use balls jars in a regular stove top pressure cooker and you can simply cover openings with aluminum foil. You don't need such a large pressure cooker, and most people try to use a microwave because of the convenience.
Great video. Limited applicability for sur, since you need to A: heave firly "valuable" or rare plants and B: want A LOT of them, but I would love to give this a try. I mostly grow vegetables, but the experimental factor of cloning plants in vitro rather than traditional media has a lot of appeal to me!
Tissue culture is huge in the agricultural industry, even for non "valuable" plants!
It's interesting that you use the colorized "pH Up" and "ph Down." I generally reach for phosphoric acid for hydroponics applications to keep system complexity down. Also, the Presto might be worthwhile to have but I was using the InstantPot for brewing and other zymaturgy applications for some time as its automation is just so good.
A simple suggestion, On your still air box, glue, two six-inch Outside diameter PVC pipes, cut 1/2" length, to the outside of your still air container around the arm holes. Then use veterinarian examination gloves which have long sleeves for the examination of cattle. Each can be put through the sterile hole and rubberbanded on the outside of the pipe which gives you an even better chance of keeping out skin cells or outside contamination. A fairly cheap and inexpensive way to add another DIY level to eliminate cross-contamination. Exam gloves can sometimes be acquired from a local co-op.
At that point you’d be making a glove box, which if I’ve educated myself correctly is used for a different purpose than a still air box, namely working with things that need alternate atmospheres. If you can fully sterilize the air in the container then the glove box is the way to go but for most folks allowing the physics of the still air box to work should yield more consistently clean results. You get more eddy currents in the air working in a glove box than a still air box, so if your air isn’t sterile a SAB should treat you better for tissue culture. I could be wrong though
Excellent !
Very informative 👏
Thank you for the info! I've been curious about tissue culture cloning for my aquarium plants like bucephalandra and anubias. I recently started learning mycology and theres a lot of overlap in techniques and equipment. I lol'd at the cannabis remarks.
Its very educational! I wish you can also show the process of corn (maize) tissue culture. Thanks
Nice video. Learned a lot.
Are you going to do a video on introduction of these plants to soil? I've always had issues with soil shock and heartiness going from hydroponic to soil.
I have an older video on the topic that I plan to remake soon :) I usually add a little vitamin b-1 when I water freshly transplanted plants and I spray them with wilt stop if the leaves start to wilt
i am hooked!
Gracias, muchisimas gracias.
I grow cannabis and it is the only reason I am watching your video. That being said I love your video and really appreciate the information! Attagirl
This is so cool I really wanna try doing this, Thank you so much for your videos!!!
I'm glad the videos are helpful :)
Thanks a lot for sharing.
Up to now, only succeded from orchids seeds but never from leaves and stems cuttings.
How you sterilize your leaves?
Awesome video and thanks for all the content, I've been learning a ton from your videos. I also have a question that you could possible answer for Q & A video (or it might make a good video on its own some day as its a fairly packed question).
How would you go about germinating old seeds? What method would you use to germinate them and how would you sterilize them. What media would you select for the old seeds to grow in? Is there a way to sterilize seeds once they have germinated or will all sterilization methods hurt root radical?
If you wanted to germinate seeds in tissue culture, you would need to sterilize them before they germinate. Seeds can handle a pretty high concentration of bleach to clean them. I remember reading that UCLA germinated 500 year old lotus seeds successfully, but the viability of seeds is usually much shorter than that, so it probably depends on what you are trying to grow. You could also germinate the seeds in soil and then put a tissue sample of the resulting plant into tissue culture, which is probably what I would do!
THIS IS SO COOL! is there any plant that can't be micro-propagated?
If you could also discuss ways to make money doing this hobby that would be so awesome.
Great vid btw!
Keep up the good work 👍👍
Hello, you convinced me to get started ! THOUGH I've only seen videos where a baby plant is going in the biocoupler, but can you use a leaf cutting, or a stem like with "traditional" TC ? thank you !
18:25 best answer ever. Love your videos i am just beginning to look into tissue culture after many years of doing cuttings, Dont get me wrong I will always love cuttings but mt answer for why tissue culture would be the same as yours
👍 Thank you!
I like your videos, good information presented in a way that is easily understood but not overly simplified. I am just starting to research this, and my question is if there is a time where you can suspend growth and save for multiple growing seasons? I know they freeze embryos, is there a way we pause the process? Thank you.
What would you use instead of PGR's if it's edible plants you're growing?
After you clean your steel air box if you wet the sides down with soapy water it will work even better. The idea is that the drops of soapy water catch dust particles before they fall onto your clean work. If you do choose to use isopropanol be very careful not to have any open flame nearby because the vapors can catch on fire
Thanks!
Thank you so much!!!
I like ur videos content so much
Would you be able to use an instant pot with a pressure cooking setting? Would it need to be one of the ones with an interchangeable lid for the setting? There's some options around 50$, would cut the price in a third
Thank you.❤
Tissue culture on cannabis seems to present many new challenges, refer to cannmed chanel for some interesting talks on that.
TC on cannabis is mainly used to clean a plant from hop laten viroid, and you must only cultvate the meristeem with much care abour leaving the vascular parts out of the sample.
Great vid series
Were can I buy biocuplars in South Africa
For pressure cooker sterilization you can also get away with using an instant pot if you already have one. You just have to see in the manual what pressure it can get up too (usually around 10 or 12 psi) and essentially just cook it longer.