Fluorescent microscopes are amazing!
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- Опубліковано 13 бер 2019
- Fluorescent microscopy was always one of my favorite parts of working with mammalian cells as it always made for spectacular images. There are a variety of techniques that make use of fluorescence microscopy, many of which are used regularly in the best labs in the world. But fluorescence microscopes themselves come with a steep price tag long before you think about reagents.
Over the last few months I've been exploring how to convert a cheap OMAX entry level microscope into a fully functional fluorescence scope which I'm happy to report was a huge success. The mod uses a 5mw laser and a special but readily available filter and produces stunning images on par with some of the commercial models.
This is the filter I used: www.ebay.ca/itm/RGB-System-Op...
If this link stops working just search "reflect blue pass green and red" on ebay and it should come up. Currently it costs ~5 dollars. When I recorded this script I thought it was ten. So total mod cost 7 whole dollars. 5 for this filter, 2 for the laser.
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_____________________________________________________ - Наука та технологія
You took everything I knew about biology, threw it out the window and gave me one of the most interesting sciences there are.
In case some don't know, if you have a low to medium end DSLR of the CANON brand, then there is a third party firmware called 'Magic Lantern', which adds many features found in much higher models.
but are any of those features relevant here? wouldn't the limiting factor be sensor quality?
@@nicktohzyu probably, but I'm more thinking about the potential of using timelapse and/or the hdmi out not enabled by default [on lesser models].
People like you are why I go on the internet. Thank you.
Brainbow does not label different neuron types with different colors. It instead labels a set group of neurons one of 4 different colors due to a stochastic recombination event occurring in each cell (each cell that will eventually glow starts out colorless and then a really cool genetic tool randomly picks a color and then remains that single color forever). This gene can be inserted multiple times giving the neurons the possibility of having multiple random colors selected. Then by imaging the brain in all 4 colors and then calculating the number of each copy being express by each cell(using differences in intensity to differentiate a neuron with two green genes and one red from a neuron with one green and two red for example). By individually labeling the neurons with these random color sets it is less likely that two identically colored neurons will be right next to each other. This lets you see the structures of these neurons.
This is my different than labeling different cell types (for example having green pyramidal neurons, red PV positive Interneurons) because each of the cells of a single type would have the identical color.
thank you for your hard work and brilliant videos
This is amazing! You have been a huge inspiration for me for quite some time now, broadening my horizons and showing how much amazing equipment and devices can be build in one's workshop. Cheers and keep doing what you're doing! :)))
Wow! I know i found my forever favorite youtube channel! I love how you get in depth with details! You post very simulating content! Keep doing great!
wow, congrats on the budget build, glad you were able to save so much money, cant wait to see what you do with this :)
Thanks for putting these videos out. I have tried a simpler process to get fluorescence microsopy and it has worked incredibly well. I replace one of the eye pieces in the binocular ocular with an LED source, that is, LED light enters the "eyepiece" and is focused onto the sample by the standard optics. The camera is attached to the other eye piece. There is some reflectance of the LED color which needs to be blocked by a filter (on the camera lens in my case). I have to make a youtube about this - hopefully in the next few weeks.
Keep up the awesome work.
Nice work!
If you could control the focus digitally somehow, either by interfacing with the camera or motorizing the focus on the microscope with a stepper motor arrangement.This would enable it to focus in and out over a set range and take images repeatedly over the entire focus range of the sample. With it being automated you could even do long exposures. You could then use focus stacking to get an image of the entire sample's depth in focus.
Thanks a lot !!
I've used these in medical research labs many times, but I somehow never stopped to think about the actual engineering side of it. Fascinating.
To solve your focus problem I suggest that you act by trial-and-error : the focus point of the two wavelengths are always at the same distance to each other so if you know the position of one you can determine the position of the other by using the experimentally discovered offset. To speed up your search space use binary search.
there are uv leds that work around 365nm than the 407nm region and may have a less significant bleed. also, have you tried filtering out unwanted wavelengths prior to exposing the sample?
太好了!感謝你
Neat-O, as expected
First of all, thank you for being amazing and making such great videos.
Would it have been possible to just shine the laser in at a 90 degree angle to the optical path and reflect it up with a 45 degree mirror instead of repackaging the whole thing?
How long of an exposure did you need? Looks amazing.
Good work. Try epifluorescence approach. I got some decent results just by using that.
Hi, noticed we have the same microscope! I was wondering what camera adapter you are using as I am having some trouble picking out mine (have a micro 4/3 camera instead of a DSLR, but any info would be great). Thanks!
Question: Have you considered something which diffuses the UV from the laser enough so it is more like the original LED in the microscope?
Would remove the hassle of having to arrange the collimator lens, just pop the UV laser in the right place, maybe even have both the white LED and UV one in place?
> budget research lab with non standardized and non health regulation approved methods
I wanna try it 🤔
It's not that far off from how things are done in actual academic labs. One of the people who started work in the lab I did my post-doc in remarked that she had always thought of labs as sleek modern places with new, gleaming equipment. She was surprised to discover that in reality most labs are cobbled together, with duct tape, wire, and glue holding everything together.
5:57 you will get way better diffusion if you add a diffuser filter between the lens and the led and if you leave a gap between the added filter and the lens
Excellent presentation, although my big question is directed towards safety or prevention, since the risk of contracting skin cancer (melanoma) is quite high, since not just any plastic insulator is worth to protect yourself. Regardless of the fact that vinyl is an excellent protector, what filter do you use to protect yourself?
UV protection glasses are also very important.
A fluorescence microscope is very expensive, but it offers an isolating chamber where the conjugation of ultraviolet rays to the slide is safe.
Many times one has to wonder if it is worth seeing beautiful images knowing the great risk involved.
All the best.
It might be a better idea to mount the entire laser horizontally and use a mirror instead. (lense after the mirror)
That way the laser´s housing can still help cool the diode. I believe regular mirrors should reflect the UV fine.
You could place a visible light blocking filter infront of the uv light
You are very smart :)
is there a way to get those yeast cells that express well?
You should get software for your camera so you can see your exposure on a computer screen as soon as it is taken.
Maybe you could dial in your focus that way?
I know Cannon camera's come with software that lets you control focus. iso, exposure... Almost every function really
i'm not sure about your model?
Really awesome pictures i'm sure they will only get better moving forward best of luck
Can one change the light source of a regular microscope and use the microscope for fluorescence work.
What about using a UV source on a dark field microscope? It wouldn't be a total loss of pumped light, but would it be partially viable? Thanks for your great work!
That technique (transmitted darkfield UV) was used decades ago and worked ok. It was replaced by "direct fluorescence" which just worked a lot better. This is a technique where the beam of UV light is directed from above the objective lens onto the slide using dichroic mirrors along with objective lenses designed to pass UV light.
You can find parts or whole sustems for either technique on ebay though it can get pricy even for old obsolete microscopes. Having said that, if you can afford to spend that kind of money on your hobby, the results can be spectacular.
What do you think about AuREUS Solar this from this Philippine student?
Have you tried cooling the laser junction? This would let you run the laser harder.
Can this perhaps be used to colour different species of algae in different colours?
I've got a microscope ad on this video
Could you please share the RFP plasnid for ecoli?
We are happy to see your passion on fluorescence microscope. Welcome to study fluorescence microscope with LED light source for Carbon neutral society purpose.
you might have a little bit better reliability with your laser mod if you current limit the power source, rather than voltage limit
If you focus with visual light, it will be ever so slightly off in UV. For Near IR photography, the lens transmits broad but the medium(film) on reacts to IR. For digital near IR you open the sensor to full CMOS range but cut away everything visual and keep IR. However. On analog cameras. You focus with Visual, and IR is a lot off often. Best is to build a reference scale due to the lenses not really made to correct chromatic abbortations beyond visible light.
Could a future project be to build an optical bio-photons scanner? To measure the quality of the food we grow? Maybe it is way to high-tech but it would immensly help people out to check if the food they grow is of good quality or not. And yes, I am serious.
can you please upload the upper green photo at 0:31?
I have an OMAX scope and I use it in my fish room and I like to make videos about it. I would like to figure out how to make my OMAX Fluorescent.
We need to protect this man, and make sure he procreates.
I support this message.
Woooow
Why don't we use di-coloric mirrors to enhance laser safety glasses?
I wonder, could you use that to select single yeast cells that express the fluorescent gene the strongest, and grow them into super bright colonies?
It would be exceedingly difficult with this setup. That's getting into pretty hard core microfluidics at that point. But some styles of flow cytometery do exactly that.
to fix the visible light bleed, couldn’t you just put a UV bandpass filter between the UV LED and your sample? that way most of the visible light is blocked and you won’t get that greenish light contamination?
Now if you can just make a phase contrast mod for basic scopes.
Do you have any onesBilly in the housing
Link for lazer?
Maybe try using multiple lasers.
You should take a look at thermo-fisher scientific.
If you want better imaging, i suggest the sony a7s.. the first one is good enough. You will have over 32000 iso clean image.. even 205000 iso is quite good
If you send me samples i can put them under my florecent microscope
Please make modification on microscope to watch zno nanoparticles if not nano then 500micron m. Nano flower
Ever thought to try a cheap USB microscope for the image pickup?
Also something unrelated, I was wondering are you able to program cells to build different proteins based on a trigger lightwave length.
Just a wandering thought I had last night. (I'm engineer not a biologist :-D)
One of the items I was supposed to recieve a while ago was a plasmid that makes bacteria light sensitive. It's called pDAWN. on blue light exposure it will turn on protein synthesis. In that case it was a version that produces RFP (same protein in this video). Its slow though. You need to expose them to light for like 24 hours to get a good color change.
@@thethoughtemporium Well the reason I asked because I had the idea of using a "nutrient-slime" as a carrier medium for a variety of different cells/organism and light projector to build a organic 3D printer.
Could be interesting to use it to build medical scaffolds or even entire "replacement" limbs if the process is sophisticated enough.
@@DerSolinski Cool idea
What you're describing so is incredibly difficult to actually do, it's really hard to convey just how difficult it is.
@@thethoughtemporium 😎 difficult ~= impossible
So now I just need to persuade someone to devote his life to this project, how hard could that be 🤣
Hello good afternoon.
I find your videos incredible, I work with biodigestion of organic waste, composting and biofertilizers. one of the things that intrigues me most is microorganisms. They are faciantes.
I am agronomist, I have thought about the possibility of working improvement microorganisms In to accelerate the process of decomposition and transformation of nutrients. Do you think this is possible?
Thank you, Dr.
In theory, can you make anything fluoresce with the right lighting? I mean, even non-fluorescent things?
Imagine humans glowing when they get excited!
I mean, sure, technically. You might just need an xray beam to do so.
How stupid am I, I wrote the last question before I watched the video. This is exactly what I want here.
When I was at school, I switched doing Physics and Biology to do Chemistry and Biology because Physics wasn't "compatible" with Biology....... how wrong I was.
If fluorescent microscope manufacturers could just use this technique to make cheaper scopes.
came here hoping to find out how to turn myself fluorescent smh
Would a ZWB2 filter help? budgetlightforum.com/node/41697
Can you translate it in Arabic language please 💗🤗
Blue ray lasers are dirt cheap and 405nm
Good video but those IR lasers are dangerous as fuck. They can make you go blind in milliseconds.
very unsharp and never never use your eye with blue laserlight even with filters. why dont you use epi-fluorescence even your laser is bad aligned
Do you know much about atomic gardens? Is it possible to get jimson weed to produce cocaine via a genetic mutation from an atomic garden? Jimson weed already produces atropine, which is very close to cocaine. Can i take apart old smoke detecters to make an atomic garden?
Be careful with those cheap lasers from China. They are often significantly more powerful than advertised as.
Hey, could you make a video about this paper? ve42.co/magneto it's about the ability to sense magnetic fields and veritasium made a recent video about it. For me it seems a little sketchy so it'd be cool if you could validate or disprove these claims based on the data.
In the veritasium video the experimenters stated that alpha wave activity goes down when a stimulus is present, such as when paying attention to something. Couldn't the counter clockwise and clockwise rotations just be random moments of people paying attention to something else? It seems like this is no different from random chance to judge whether or not you are picking up magnetic fields with just the alpha wave activity or it could have a false positive.
Dude your shit isn’t focused!!