Building a Nanodrop Style UV/Vis Spectrometer
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- เผยแพร่เมื่อ 7 ก.พ. 2025
- 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.
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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 ^_^' )
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
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!
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
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
...and don't forget that OD is a logarithmic quantity
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.
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 ;)
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.) ;^)✓
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.
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.
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.
You're videos are great, not only for the actual builds, but for the absolutely superb explanations.
I’ve been looking for this tutorial for months and didn’t even know it. Thank you so much.
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.
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.
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 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
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.
The world could use more scientists like 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
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 just love your videos 😍. Waiting for the spider silk video series
Sounds nice!
What a prize to find this channel. Thank you for sharing this great content!
Underrated video, honestly
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!
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!!
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.
AMAZING! can't wait to see what you use it for!
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.
Very nice, I look forward to seeing the additional modifications to this design.
Lovely video mate! keep it up, big support!
You are the smartest man on the Internet. I used to be the smartest man on the AOL years ago.
WTF thats absolutely amazing wow
You can measure DNA with absorbance at 260nm, don't need fluorescence if sample concentration above 5 ng/µL or so
This thing is amazing! I can't wait to build one :-) Thanks for sharing
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.
I didnt know There was a software to do that!
As always nice video!
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
your videos blow my mind every time
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).
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!
Brilliant video! Loved it thoroughly.
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.
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.
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.
Yesssssssss another upload ❤❤
@8:36 "On a microscopic scale, a diffraction grating is essentially a series of tiny prisms"
No, no it is not. Refraction is NOT diffraction. They are 2 completely different wave phenomenon. Refraction is light bending towards the normal as it moves from a less dense medium to a more dense medium (or vice versa) because the speed of the wave is different; diffraction is light recombining into a new beam vector based on the distance between the holes in the grating. And diffraction gratings are on the sub-microscopic scale, they're on the order of the wavelength of light, you're not going to see the lines of a diffraction grating in a microscope. You sounded so promising before you said that...
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.
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.
Near Slit , What is there ??? Diffraction grating or Mirror ?How to select wavelength for this caase? for example 400-700nm for analytical analysis of chemical solution or dye /Where you had connected DVD Diffraction grading? Have you connected the mirror? At the edge of the slit? Or you have connected in front of the camera-Lens?How to get absorbance -wavelength curve for a cuvette
This really should exist as a build kit with precut mirrors and stuff...
Black hot glue is fantastic for sealing light tight boxes.
You could accumulate data over multiple frames of the video in order to get more precise results, it's not like there's much if any external noise here, so your exposure time could be arbitrarily long.
I like biology but your videos really want me to go into the field
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.
Well 10k dollars for a spec that can measure DNA.....well i have the exact same problem with identifing metal alloys. A handheld XFR is also many 1000's of dollars...Can you make this work to detect aluminium alloys for example. One would need to "excite" the alloy somehow, but it might be possible.
0:52 Raises the question: how is it that the Sun is seen as white? In fact, it's taken as the primary reference for white, isn't it? ~~~ The question triggers all my understanding of color psychology and developmental psychology. The best answer I have at this point is that we get wired that way when we grow up going out to play. ~~~ Well, there's a question that's going to have my little gray cells occupied for the next couple of decades.
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.
This project is amazing. My question is: can I measure the soil's nutrients with this device?
Upwards of $100,000 for a spec that can do DNA? I guess I got to play with some pretty expensive hardware during my time as a lab tech/intern.
feeling smarter. thanks
Wisdoms more important than knowledge - Einstein
An I'm a master of wisdom""
are just feelings
Хочется, увидеть спектр обычной лампы накаливания ;)
Только он может показать точность измерения спектра.
Great video -- just a note, Black 3.0 is now available and is way blacker!
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)
AMAZING! Thanks
8:43 1000 lines per mm... What are the other line counts you tried? Are there higher counts? ~~~ Now I'm wondering what it would take to make these at home, as well as how to ensure the highest "geometric" quality of the physical grating.
Cool to see another subber to BenNbuilds
Excellent!
This is pretty awesome :)
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.
Nice! I think next is a flow cell and a pump and some silica for a column
How important is having a really high quality diffraction grating here? Is that cheap 1000 line the best possible option? Would a glass 1000 line version be any different? How does using a CD change things? What does the diffraction grating component in the project impact beyond focal length?
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.
The amount of light absorbed is also affected by the distance light travels through the medium( the thickness of liquid on the glass). Your equipment would be more accurate if that variable was controlled.
The original design is done to consider this, based on the original nanodrop. For some reason he discarded that.
Pretty interesting video. I've used NanoDrop 2000 to measure DNA concentrations.
this is so cool!
Is it possible to calibrate it with commercial calibration samples to measure some substances within eaxact ratios, like sulphur in appel juice, potassium in water?
I would think so,
You might could make your own calibration standards.
Like maybe make 3 known concentrations of potassium chloride, then use that to extrapolate out what the potassium would be in sampled water.
Or maybe use ammonium sulfate to get the sulfur concentrations
@@Abdega thank you, it is worth to give a try
Technically it's a spectrophotometer, not a spectrometer, since it deals with spectroscopy, not spectrometry...But still, you're a genius.
Dude, you are amazing. Thank you.
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.
Thank you so much
You should rent a high end Spectrometer and make comparisons, could be really cool and give an idea on if you can get away with DIY.
In a Nanodrop, the light source is a xenon bulb, unfortunately a LED don't emit in UV-C (230 nm, 260 nm and 280 nm are important wavelengths in UV-Vis spectrophotometry), so you can't measure the absorbance of nucleic acids and proteins.
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.
I have done some study in the subject. It is a fascinating thing, Light and color; The way our eyes see color and how our brains perceive it. But, if our eyes and brains could see what really is, we wouldn't enjoy it very much. Luck to be human.
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 :)
Very clever.
Great video. I’d like to build this to measure the color of vintage postage stamps. Do you have any advice for doing this?
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?
Love ur vids!
Dude you should totally make a spectroheliograph
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
Don't you guys have a 3D printer? Back in the end of the 90s (no 3d scanner then, mostly lego) we took a scanners ccd sensor (usually about 2048 pixels in a line) and collimated a spectrum onto it. This might be more work but ultimately should lead to a much better spectral resolution than the webcam (in that crop, how much do you have? 500 pixels?) not sure about how to get it into such a software though, maybe talk to the authors to enable them to write data input plugins.
You've got an amazing educational system going for private research for the general public. I'd like to discuss working together sometime.
11:10 why would the infrared light damage someone's eyes? since it's at a higher wavelength (ie lower frequency ie lower energy level) than visible light, it should be safer than visible light at any rate, no?
No. Just because we lack receptors to see it, doesn't mean it can't enter the eye and cause damage
Try to rub a water on the crack lines next time before you break the glass. It helps.
Hi for linux, you can use LPUWS.