Building a Nanodrop Style UV/Vis Spectrometer
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- เผยแพร่เมื่อ 17 ม.ค. 2019
- Spectrometers are one of the most ubiquitous tools in most labs because an enormous amount of information about a substance can be gleaned from how it interacts with various parts of the electromagnetic spectrum. But for as common as they are, the price tag on a new spectrometer can be massive.
Today we go through a bit of theory on the electromagnetic spectrum so we understand spectrometery, and then build a little nanodrop-esq spectrometer for use in the lab. While it's still a work in progress, it's more than sufficient to preform many basic tests and experiments.
Applied science Zeeman effect: • Zeeman Effect - Contro...
Hackteria: hackteria.org/wiki/DIY_NanoDrop
SVG file of design: github.com/FOULAB/DIYnanodrop...
Webcam: www.amazon.ca/AUSDOM-1920x108...
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Thanks for mentioning me! I watch all your videos and really like your explanations.
Hi there
Dude you, the thought emporium, and Nilered need to start your own online University!
Ben has permanent youtube face from his last video!
Physics, engineering and chemistry!
@Barry Manilowa All absorbance is non-linear! :D
See en.wikipedia.org/wiki/Beer%E2%80%93Lambert_law
Our perception of color operates on spectra in a different way though, causing us to sometimes perceive a change in hue like you described (not just saturation/brightness), a phenomenon which is called en.wikipedia.org/wiki/Dichromatism :>
(but since you mention adding acids and bases, I wonder if you're talking about pH indicator instead, which actually changes its color (from a chemical reaction), not just appears to change color due to a change in concentration)
(but lets not hijack this thread and pester Ben everywhere he goes, heh ^_^' )
Hands down this is what diy engineering channels should aspire for. It's safe to say I have waited for a channel like this for a decade, now. Subbed and Loved!
As a guy that's responsible for training new users in flow cytometry, I used to have this elaborate presentation I would run through, now. I just link them to the start of this video. Excellent explanation of spectrometry and very succinct
Just to remind people that hot glue exists in black for this kind of application.
Must be some *black* market stuff cause I've never seen it in my life.
@@maracachucho8701 I think I can't post a link here, search on amazon "black hot glue"
@@maracachucho8701 It's also on ebay but smells very toxic
I had black hot glue a long time ago and it had a pitch/tar smell to it. I think it was for different application than typical hot glue :P
You would have saved me a whole lot of time and frustration if you had told me this back in 1992. I made a robot that used IR remote control receivers as range finders. Back then, the receivers were in little sheet metal boxes. They worked on the breadboard, but not when hot glued into the robot. The hot glue I used acted as an IR light pipe. Any IR that hit the hot glue blob was carried into that little metal box.
You're videos are great, not only for the actual builds, but for the absolutely superb explanations.
No time to follow this through right now but I had to compliment you on the excellent introduction. Definitely will come back to follow it to the end! Thank you for the quality contribution.
I’ve been looking for this tutorial for months and didn’t even know it. Thank you so much.
i was watching Applied Science video and you video was "Up next" and thats how i found out about your channel and i am subscribed to you ever since ;)
Actually you can estimate the concentration of bacteria by how much light is dispersed, not absorbed. That's why it's called OD instead of absorbance. Awesome video as always by the way
Well, both actually, you can either use nephelometry or turbidimetry
What a prize to find this channel. Thank you for sharing this great content!
cool stuff, your diversity seems vast. btw "Applied Science" certainly is/has been a great channel, Also "Tech Ingredients" is top notch too, just in case anyone wasn't already aware & is interested.
I’d like to add Marco reps and zenodilodon to that list. Especially if you were interested in lasers, spectroscopy, and precision lab equipment
Very nice, I look forward to seeing the additional modifications to this design.
Nice.
I found this a bit funny since, just yesterday, my physics teacher showed us your video on measuring Planck's constant using light diffractions.
Wow! This is incredible! What a nice project I'll have to make :P
I'm so glad you posted this, thank you so much!!
AMAZING! can't wait to see what you use it for!
This thing is amazing! I can't wait to build one :-) Thanks for sharing
Lovely video mate! keep it up, big support!
Brilliant video! Loved it thoroughly.
Nice build and presentation. All systems started with a handmade system for the most part. Increasing accuracy as needed with improvements in tools to make improvements in whats made and so on.
I am a MSc in Biotechnology and I am still learning cool stuff which I could have done thanks for sharing such goods information for free.
If I were ever to be marooned on a deserted island with only a can opener, a flat blade screwdriver and a flashlight, I would hope to have you or your doppelganger with me!
The quality of your work and narration is simply stunning. I really have hope for the future of humankind when I see that the world is awash in smart people like you.
May you find peace and prosperity all the days of your life. Amen.
(Just a quirky little thing I picked up on TH-cam.) ;^)✓
your videos blow my mind every time
I've used the 'proper' UV-vis machines at my university (I study chemistry), and this was really interesting to watch. I might have to try and build one of these at some point
I didnt know There was a software to do that!
As always nice video!
The world could use more scientists like you :)
I just love your videos 😍. Waiting for the spider silk video series
Sounds nice!
Underrated video, honestly
Dude, you are amazing. Thank you.
I love the idea of making a hyperspectral camera that gives 4D data and the ideas behind it are very interesting.
Goddamit, i wished i had hyperspectral vision, that shit was awesome, btw i.m searching a way to use a high speed camera to do exactly that, there's a group that uses a LCD polarizer to change the phase differential and create a railleight filter
Thanks for the inspireing video!
Looks like I am not the only biologist affected heavily by phisics and electronics.
I am curently building a spectrometer to test the light sources I am going to use to grow my plants.
One note... you actually try to use your "nanodrop" in a fluorimeter mode. It is challenging using the current geometry as excitation light is usually orders of magnitude (3-4) more intense than the emitted fluorescent light. Ususally the emission is measured from a perpendicular direction, so the excitation light does not hit the detector.
You should be able to measure DNA based on absorbtion using 260nm of full spectrum light (e.g xenon flash): A(260nm DNA) - A(260nm Water)
To split off narrow strips of glass I recommend using a pliers that has vinyl, masking, duct, or painters tape wrapped over the pincers. There's actually a couple special bronze tools for splitting glass used by glaziers, but they're all basically variations of pliers. Putting Kerosene or alcohol on the glass before scoring it can help create an uninterrupted scratch which breaks cleanly. It gets more difficult and less forgiving the closer the strip being cut is to the thickness of the pane.
Glass is surprisingly amenable to sanding, and the edges could be sanded flat without compromising the strength of the piece. However, sanding the edges might cause back scattering of light and mess up the function of the spectroscope.
Actually it's pretty simple to cut it using a rotary tool and a diamond blade.
i love this scattered sciencey stuff
Their is SO much more u can do urself
Astounding
@@SystemsPlanet read beyond the light barrier
@@SystemsPlanet as in scattered topics he covers
AMAZING! Thanks
WTF thats absolutely amazing wow
This is really amazing. I'm learning about spectrography. I'd like to make my own to analyze grow lights for indoor gardening. I'm confused about a few things. You use a white led in your device, but my understanding is that white LEDs do not emit very broad spectra. In your computer program the image showed a very even spectrum. Did you use a special LED? Also, I've looked at many DIY spectrography, and one thing they all seem to neglect is the fact that the camera isn't equally sensitive to all frequencies of light. I haven't figured out how to calibrate this away, given that I can't easily generate light with both a known wavelength *and* intensity.
Yesssssssss another upload ❤❤
Now I'm sure that TH-cam can read your mind. I was thinking of Thought Emporium's DIY spectroscope and this video showed up in my feed right away.
Something I've always wanted to do see the other portions of the spectrum. There's such a world we don't know about
Lots of good ideas here. I would like to add that finding a ready made black box would make the manufacturing part much faster. For example, another builder used old VHS cover case made out of black plastic. Hot glue should stick that pretty well and you can easily make the required holes on the plastic.
Excellent!
Nice! I think next is a flow cell and a pump and some silica for a column
This is pretty awesome :)
Thank you so much
Wow, i really like your videos, especially this one since you mentioned works of ELISA tests. as i was misdiagnosed with stage 2 Lyme borreliosis, i did an elisa with a false positive and had to bear with 2 years of therapy for lyme disease. and elisa was veeery expensive at that time, it probably still is.
I remember I made a analog paper recording model, with a motor turning the grating, and a couple of light sensitivite resistors! It was just weird, compared to the instruments of today, 40 years later!
Hi, are there any updates on the Spec? I am probably going to try it out. I am especially interested in the identification and quantification possibilities :)
this is so cool!
Nice job. I wanna know, what the type sensor u use??
You should pick up some black hot glue. It is amazing for projects like this, where you're trying to limit light coming through cracks.
Very clever.
I can make this. Thank you!
Love ur vids!
tip: to get a surface reflecting mirror.. rub paint thinner on a regular mirror.. it strips the color coat revealing the mirror surface.. hobby stores sell small precut mirrors..
I just rather use HDD platters, mirrors corrode over time and HDD platters are perfect mirrors that don't cost anything.
This kind of tech is how a lot of water testing is done. Colorimeters use reagents that change color in the presence of certain chemicals, and using a filter that selects only for reagents color, you can quantify how much of the detected chemical is in water. You’ve done something like this when you mix purple cabbage juice into a liquid although in this case, it’s the PH your detecting.
i know that styropyro has a pure uv laser pointer
but the diode he used was a nichia NDU4116 , 375 nm laser diode but that thing costs like 4300$
or if my calculations are right a 750 nm lazer diode sent through a non linear cristal shuld work too
Damn dude if you know a lot about that stuff, TH-cam sure as hell could use some more quality laser/science content. Don’t get me wrong styroPyros channel is cool but I am much more fan of the science oriented stuff. Like this channel, and Marco reps, zenodilodon, les’s lab, Tech ingredients, and others
Pretty interesting video. I've used NanoDrop 2000 to measure DNA concentrations.
Love it!
Could this work for telling how much UVA, short-spectrum UVB, long-spectrum UVB, and UVC is emitted from some bulbs? (At least assuming the bulb doesn't emit much heat)
I hope someone releases this as a kit
Could be usefull for all sorts of things
That's what's shown here, a kit with some additional modifications. There is a link to the kit in the video description.
I would like to know how you get the thermino spectrum tool to work with a narrow spectrum? I can't ever seem to make it work it seems to expect the blue to be way at the left end. Great info here. Thanks.
feeling smarter. thanks
Wisdoms more important than knowledge - Einstein
An I'm a master of wisdom""
are just feelings
You can measure DNA with absorbance at 260nm, don't need fluorescence if sample concentration above 5 ng/µL or so
Very interested in Updates :D
Great video. I’d like to build this to measure the color of vintage postage stamps. Do you have any advice for doing this?
your contribution in this field is mind-blowing but I have a query.
What was the total cost of febrication?
And which software was use to peak display
Hoya makes a great filter that removes most visible light and only allows ultraviolet light to pass. I use it for viewing florescent minerals under UV-C shortwave 265 nm light. It blocks out everything but the UV-C perfectly.
I like biology but your videos really want me to go into the field
Awesome! So in what spectrum range are cheaply available sensors sensitive?
Can you remove the beyer filter to get a monochrome image with higher resolution / sensitivity (throwing less light away and less noise).
Thanks for the video your channel is really awesome!
Lots tried, you have only some luck amd throw away other semsors you mistreated by rather some chemical-mechanical brute force approach ; it’s a pity real b/w or UV or NIR photography has not more friends.
There are people selling modified camera bodies with abrased filter layer, best re-seal.
You've got an amazing educational system going for private research for the general public. I'd like to discuss working together sometime.
To quantify DNA (or anything really) you will also need to calibrate the distance between the coverslip and the fiber optic cable end on the detector because of Beer's Law you need to know the light path length.
Using paint remover, one can turn any regular mirror into a first surface mirror. It's cheaper too.
If you don't have glazier pliers, you can use cardboard and regular pliers.
Lubricate the glass cutter or it won't make a good score: automatic transmission fluid works great for this. It takes practice to cut glass. I have a friend that's a professional glazier who showed me how to cut glass. Wear gloves and sand down the sharp corners for safety. My friend used a belt sander.
This really should exist as a build kit with precut mirrors and stuff...
After scoring the glass, sharply tap the steel ball end of the cutter ( on the opposite end of many glass cutters) directly under the scored line. When properly done, you will see an internal fracture start. Continue tapping ahead of this fracture.
I can't find that Thereamino software. Is there a link? I did find an open source general tool called Theramino, but it's not the code we see here. Great job here! I want to build this, but fear that the software will be the hardest part. It's worth the effort though.
Next time you need a mirror or glass: scratch it, heat it with boiling water then dunk it in ice water. After a few times it will break on the scratch line. Awsome build!
please make a comparison video between diy and commercial spectroscope, and i have some questions: how do you measure and control the thickness of the sample? and need more details on setting up the camera and its position and lastly, how to calibrate and start with the software? TIA
Far-Red is possibly the most neglected portion of the electromagnetic spectrum. It consists of the portion from 700nm to 800nm, roughly. The visible spectrum and infrared do not have a clearly defined boundary, and far-red is a sort of transition zone between the two. Far-red light is visible, but it is not as bright as light in the rest of the visible spectrum. Far-red will also reflect off of most of the fabrics we call black. Some of you may have noticed that many black, fabrics have a very slight red tint to them when looked at under sunlight or incandescent light especially. This is due to far-red reflecting off of them. I have some 730nm LEDs, and I built a flashlight from a 3 watt one. When I take this light into a room and shine it on my black fabrics, the fabrics shine back red. Even more interesting is the fact that if the black fabric had been written on with black permanent marker, the writing becomes readable. Far-red LEDs make any other red LED look a bit orange by comparision; these far-red LEDs are so red that no screen can display how red they are. Far-red makes up some of the colors of a sunrise and sunset as well. Have you ever noticed that your black fabrics look a lttle bit 'rusty' at sunrise and sunset?
The CIE and lumens are not perfect, because they ignore far-red and say that "anything over 700nm is infrared." I have some 3W 730nm LEDs, and they are visible, so not infrared. Before anybody says, "well maybe its just the portion of light under 700nm that you see.", nope, I have a lens from a broken digital camera that cuts off all light above 700nm, and when I look through this lens at these LEDs, almost no light is seen. When you look through the cut-off filter, if you have incandescent bulbs, you'll notice a few small color details missing as well.
What a great project and a great video! I'm always amazed how a diffraction grating instantly creates a spectrum c.f. FFT et al that require time to produce the same spectrum...I know, "duh"...but it still amazes me - the universe works in parallel and near on instantaneously....man we have a long way to go!
Keep on plugging other science sites...the world is suffering from far too much BS.
Please tell us when you find a way to quantify the data. Im currently working on a similar project and it would be very nice to compare the data properly.
Muito Obrigado (thank you).
Do you guys have a video on orgon? I tried searching and didn't see your channel come up. It would be cool to see how you make a generator.
Hi I am trying to build a spectrometer to do reflectance spectroscopy of a powder. Do you have advice for this? I'd like to be able to collect the diffuse light but also measure the light collected as a function of angle.
I love you videos man
Do you have to adjust to the metal of the mirror absorbing certain wavelengths (probably UV)?
Where can I find the yellow, translucent glass at 14:11 that you were using to filter out the violet portion of the spectrum for DNA quantification..? 🤔 Did this filter ever end up working out?
Great video -- just a note, Black 3.0 is now available and is way blacker!
Cool to see another subber to BenNbuilds
Black hot glue is fantastic for sealing light tight boxes.
How to Calibrate the Device? Means ROI of Image EM Visible Spectra. Does it give emissions? Thermino?
Great breakdown to how this stuff works! loved it :)
I was going to ask why you were planning to get DNA concentration from fluorescence, as opposed to absorbance (i.e. log(Transmission)), which is what a nanodrop/any decent professional UV/vis spec would do. But then I realized its either your camera, or your laser doesn't go down to at least 260 nm, which is where DNA absorption is usually measured. I think with the laser/camera combo you have right now, you aren't actually getting much more functionality out of having two light sources.
Also, why use a UV laser, as opposed to a more broad spectrum UV LED, or at least smaller wavelength laser? That might help you with the DNA fluorescence - if you are using ethidium bromide, it absorbs most between 210 and 285 nm (for excitation at 600 nm). So even if your camera can't see those photons, it will excite EtBr without needing to jack the laser up as high as you do right now.
what dye are you using to run your DNA gels? it would be more accurate to quantify DNA if the dye only florese when it binds to DNA, but does not when it is on its own. If you could make your home-made nano drop compare absorbance in ratio (A260/A280 and A260/A230) then you wouldn't need to add any dye to quantify pure DNA and RNA
It is great. Can you make an adaptor for an old spectrophotometer as a nanodrop?
Really cool project. I have recently thought about trying to make a "Near Infrared Spectrometer" to analyse the properties of different polymers. I thought about having multiple IR LEDs with various NIR wavelenghts and one photo sensor for each wavelength. Do you think something like this might be possible?
It exists as ready chips. AMS builds them. 6point and 11point. Calibrated. Some 20nm fwhm
Could you use this to (for example) detect and identify chemical contaminants of drinking water?
did you ever modify the spectrometer with the filter to quantify DNA?
What will be the size of the slit that can be used?
Is there any specific formula to calculate the entrance slit
The "darkest commercially available" black paint seems still pretty shiny. I have seen demonstrations of black, that looked like black hole compared to the environment.
Next time , cutting glass, score the glass with your glass cutter, then hit it a few times with intense heat (butane torch maybe?). The thermal shock will cause the glass to shatter along your scored line (which is hopefully a straight line).
How do you achive the same thickness in every one of the mesures?. The absorvance also depends on the thickness of the sample.
I wanna use this but for my DIY HPLC, is there any tips for me to use some silica tube instead of manually pipetting the samples?