Verifying Serial Dilutions with the Microscope: Multi-well Breeding
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- เผยแพร่เมื่อ 21 ก.ย. 2024
- This video is the first step in our new Multi-well Breeding Technique! Enjoy and MUSHLOVE!
Amscope link
AmScope - 40X-2500X LED Digital Binocular Compound Microscope
amzn.to/3j93sLw
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I've watched all your videos over the past 4 years. I think you're putting out your best content ever. Getting people excited about fungi is just a piece of it, but providing people with an understanding of the incredible work that goes into determining the WHY is just as crucial. Thank you so much for your contribution to the field of mycology.
Gary, you rock! You creat some of the best (most detailed) mycology lab work on TH-cam. Thanks so much for sharing your passion and knowledge!
In spite of a booming industry selling spores for microscopy, you are one of the very few people I have ever seen actually looking at spores under a microscope. :-)
holding down the fort 😎💪
Your channel doesn't get enough love 💙
Amazing to think we can isolate a single spore!!! I can't wait to get into this lab work
Here is some more info that may help.
th-cam.com/video/nViO9Y4Yxfk/w-d-xo.html
3 new videos in 4 hours? nice!
On the other hand, maybe have a look at if scheduling videos, every Friday at 4pm for example, might make the youtube algorithm a little bit happier
thanks for the suggestion I have a hard time holding them back! 🙏🏻🍄❤️
You by far have the best /cleanest methods out of all the vids i've seen and I appriciate the fact that you're showing ppl proper technique. there are so many vids spreading bad practices to the new generation of mycos I cant believe it...there using SAB's with no gloves, coughing into there workspace's or saying you dont have to pc this or sterilize that..a few fails is enough to discourage one from something as amazing and addictive (Mycology I mean) as fungi over some bad tek or info so 5 star brotha you got my sub for life and will def recommend you to anyone I come across looking to get into this field...Mush Luv!
thanks homie! I just document my process - sometimes I get contams even with this much experience but I try to encourage people to start small like pftek so they wont give up
Thank you very much gary for your sharing,as usual your videos of great mycological interest,greetings for you and all the crew of fresh farm fungi,fom Algeria 🇩🇿🍄Love.
Cool video but I am curious why you chose concave slides. By doing so wouldn’t the spores be suspended at varying depths, and therefore different focal planes, potentially making them more challenging to quantify?
yes this is true, I mostly chose it because it would fit the same volume as the wells Im using. The other slides would not hold that much volume and I don’t have another pipetter - ideally you could scale down the volume of the pipette and use a flat slide and everything would be much clearer thanks for the critique! 🍄❤️
@Fresh from the Farm Fungi, I was just wondering. It would be cool to use a hemocytometer and really pin down your dilutions. But I only say that because I think such procedures are fun. From a practical stand point, as it relates to what you are doing, it’s probably not really necessary.... but would fun. I bet you would dig it. Hemocytometers are pretty inexpensive and kind handy for nerding out on stuff like this. And from your videos it does seem as though you quite enjoy incorporating microscopy into your processes.
Hey Gary! You uploaded this video 15 months ago, and I don't see any follow-up videos to this series on the relevant playlist. Did you abandon this project? Or are the videos still on the way?
Thanks for this video! I found it very interesting -- I can't wait until I can afford to get a nice microscope 😁🔬
Thank you for the informative video Gary.
Gary, how would you elevate the cap when capturing spores ?? Thank you for all your content. Love the deep dives and all the technical procedures to help us become better.
check out this method th-cam.com/video/GaO2q-mTvJg/w-d-xo.htmlsi=OARLult_IFyWCbUk
gonna show this to my bio professor and see if I could do a summer project based on this method
awesome!
Thank you for another great video! You are a great teacher!
I have a question: you mention that the serial dilution tubes are not sterile, and that this is OK because the spore print itself is non-sterile. However, one of the properties of the serial dilution is that it will also dilute the contaminants. If the final dilution tube itself harbors contaminants, those would not be diluted! Because of that, a contaminated tube seems to me to be a greater problem than the contaminated spore print. Isn't it then quite risky to use non-sterile dilution tubes?
Hi Gary my bags at first was little condensation I read they had be warm they are in my wiring cupboard on a box out of direct heat , will they dry out , there’s no condensation now look dryer it’s only been a week , thank you great channel
they should be fine maybe the first room was cooler and this caused the condensation (cooler air around warm bag)
🔥nice😎
could pipette out to slides from 10^4 downwards without changing tips for some economy
When are your live streams bro
Quick idea, not related to this video. If you have a windows PC, maybe have a look at passing your finished videos through Nvidia RTX Voice (works with all recent GTX cards too) to remove noise from you flowbench. It works like a charm for me when recording videos in noisy environments
I have a mac I will look into it
That was hugely informative, thanks again Gary!
I do have a question that you might hopefully see. If you're going to to the single spore level of detail, I would imagine that, apart from hoping it is viable, you would want to make as sure as possible it will carry a given trait you're after, right?
Does that mean this spore solution you start dilution from comes from a print of a multiple generation cloned fruit? I mean rather than use spore prints from chosen fruits generation after generation, is it not more deterministic to do this selection via cloning?
Or is it that karyogamy shuffles things so much during sporulation that it supersedes any genetic selectivity provided via cloning? I'm really puzzled and find it difficult finding any significant answer out there, everybody seems to have diverging opinions :)
the only true way to know with precision is to do DNA sequencing on selected haploids, grow them out and trace any characteristics back to the haploid culture after its paired. Then you can systematically create a haploid bank with known traits and link them to genes. Cloning will stabilize a certain trait but you can’t take that clone and cross it with another known clone to get desired traits (with a few exceptions) unless you do many many crosses and “hope for the best” if that makes sense. Watch my video on Buller phenomenon and di-mon mating - it is natures way of keeping things consistent. We, as breeders must hack this system and the best way I know is to collect haploid cultures, figure out their strong genes and bank them for later breeding. It’s a long term but powerful project.
@@FreshfromtheFarmFungi That it is! And thanks for your response.
That sounds like work for the pros, which I'm very far from being... sequencing is way out of my reach. But I think I get what you're saying. Which is that trying to pair two haploids from selected clones involves low chances of success and more crosses than I thought would be needed. I was indeed aiming for the "hope for the best" approach, seems like a bad idea!
I'll jump to your di-mon lesson, the way I understand it this re introduces diversity, increasing chances of something "taking".. hopefully I can one day get this working, at my humble level, and we'll see after that about hacking the system! 😅
Thanks again man, been following your contribution to the community pretty closely, it's invaluable.
What pipetter is this. Sorry for the spelling
Here is another video breaking down some the things Gary is covering.
th-cam.com/video/nViO9Y4Yxfk/w-d-xo.html
sure thing! good simple
video explaining serial dilutions
can you do a start to finish tutorial on cultivating cordyceps?
you have some info but not a lot of info.
#LastOfUs
yes look up hyperbreeding it’s all there
Gaz, what's up either yout scope flickering around?
Ps. Thanks for sharing