Building a Nanodrop Style UV/Vis Spectrometer
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- Опубликовано: 17 янв 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 ^_^' )
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!
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.
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
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 ;)
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.
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.
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
I’ve been looking for this tutorial for months and didn’t even know it. Thank you so much.
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!!
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.
What a prize to find this channel. Thank you for sharing this great content!
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 RUclips.) ;^)✓
Very nice, I look forward to seeing the additional modifications to this design.
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
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.
AMAZING! can't wait to see what you use it for!
Lovely video mate! keep it up, big support!
Brilliant video! Loved it thoroughly.
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.
This thing is amazing! I can't wait to build one :-) Thanks for sharing
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.
your videos blow my mind every time
I just love your videos 😍. Waiting for the spider silk video series
Sounds nice!
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 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.
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 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 didnt know There was a software to do that!
As always nice video!
Underrated video, honestly
The world could use more scientists like you :)
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
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 :)
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.
Do you have to adjust to the metal of the mirror absorbing certain wavelengths (probably UV)?
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.
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 is pretty awesome :)
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!
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.
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.
AMAZING! Thanks
WTF thats absolutely amazing wow
Yesssssssss another upload ❤❤
You can measure DNA with absorbance at 260nm, don't need fluorescence if sample concentration above 5 ng/µL or so
Dude, you are amazing. Thank you.
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?
Хочется, увидеть спектр обычной лампы накаливания ;)
Только он может показать точность измерения спектра.
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.
Excellent!
Great video. I’d like to build this to measure the color of vintage postage stamps. Do you have any advice for doing this?
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, RUclips 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
Now I'm sure that RUclips can read your mind. I was thinking of Thought Emporium's DIY spectroscope and this video showed up in my feed right away.
Love ur vids!
this is so cool!
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.
What will be the size of the slit that can be used?
Is there any specific formula to calculate the entrance slit
Thank you so much
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
Very clever.
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.
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.
Sry if this is a dumb question but would you be able to use this design for a crude IR spectroscopy as well?
Ohhh, please does anyone know what the bottom section of 2:54 is? Ive been trying to find what UV light things put out when burnt, but emission spectrum always leads me to the visable spectrum, not UV.
Something I've always wanted to do see the other portions of the spectrum. There's such a world we don't know about
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?
How do you achive the same thickness in every one of the mesures?. The absorvance also depends on the thickness of the sample.
Could you use this to (for example) detect and identify chemical contaminants of drinking water?
Nice! I think next is a flow cell and a pump and some silica for a column
Pretty interesting video. I've used NanoDrop 2000 to measure DNA concentrations.
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.
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.
Does it have a decent resolution? Can you make quantitative analysis?
Love it!
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.
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
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
Nice job. I wanna know, what the type sensor u use??
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.
did you ever modify the spectrometer with the filter to quantify DNA?
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.
Is it possible measure the light spectrum of environment light, for example, the sun light spectrum at determined time of time?
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.
I love you videos man
@The Thought Emporium Do you have the actual blueprint/CAD files for it with all the dimensions and stuff? Every wood cutting place I've contacted says all the blueprints provided here and from their main site give nothing?
Look at the interlocking "pins". They are dimensionated for the panels thickness of 3 mm plus some tollerance. Ok measuing in for ex. Inkscape is a little off but I also used Blender to remodel it with roughly 5.8 mm papel wood laminate from local home improvement store. But it got to messy so I ordered 3 mm panels. And today in addition to already received framed translucent diffraction foil I got 1000 and 13500 lines/mm sheets. I had printed grey PLA (about 3 years not in use) on the framed foil and also with paper frame removed but still I got mostly only in the corner a resemling of a refracting diffraction piece as the rest weren't a consistent surface as the foil did not lay flat. Sorry for long sentences as I am german. But my next proplem is getting the light guide.
Hello, I'm lloking for a device able to tell the exact wavelength of a UV radiation from the lowest WL (100-110nm) to the highest (405nm). Do you know if such a device exist ?
I can see sometimes UV lights that are not 405 but lower and these may be harmfull.
I can make this. Thank you!
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!
Other than just concentrations, could you use this to identify a solution by its absorbance spectrum?
Can you build a software or there is a free one to analyze the image ? Thanks
feeling smarter. thanks
Wisdoms more important than knowledge - Einstein
An I'm a master of wisdom""
are just feelings
What kind of material are you using as uv filter?
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.
hey how is the lactose intolerance going? has it popped back up yet?
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.
Hi! Just a couple of questions: What kind of resolution can you expect from a cheap plastic grating like this? Also, why do you use a white LED as a light source, given that these have quite a terrible output spectrum in terms of linearity?
You can use an LED and ignore nonlinearities because it's all just about comparing to a reference. If you pick a wavelength to measure, you're just comparing the difference between a control solution and a test solution. That's why absorbance is in arbitrary units ranging from 0-1. So long as there's a difference between the two it'll show up properly in the measurement. Unless you're doing something super finicky, this is sufficient.
@@thethoughtemporium Thanks for your reply! I am still worried though that you are pretty much 'guessing' at significant portions of the spectrum. While you can calibrate out the non-linearities to some extent, the emission of white LEDs (the cheap ones in particular) is only meant to approximate white light to human eyes, and there are actual gaps in there. You can tell theremino to cal them out so things 'look' okay, but that doesn't change the reality of there being no emission there from your source in those gaps. Also, other non-linearities of your setup might begin creeping in and wreaking havoc once you start applying all sorts of correction curves to the actual output. You're right, this is not a deal-breaker for some applications, but it is a severe limitation to be aware of. There is a reason why commercial units use broadband emitters such as xenon arc lamps. Even a small halogen would be better I think, and quite easy to implement. (EDIT:) Going back to the part of the video where you show the raw output of your grating, you can clearly see a region between the blue LED emission and the first green phosphor where there is very little light visible.
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
hey, how would you measure absorption at 200 - 400 nm?