Home
Creators
Search Recent Missing Random
Posts
Search DMs Popular Hash Lookup Tags Random
Importer
Import FAQ
Register Favorites Keys Review DMs Logout
Community
Telegram ThePornDude
Home Creators Posts Import Register Favorites Logout

DIY UV/Vis Microliter Spectrometer (Patreon)

Published:
2019-01-19 00:34:27
Imported:
2026-06
haven't archived this post yet. have a subscription? use the importer!

Content

 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.  

Files

Previews only

Building a Nanodrop Style UV/Vis Spectrometer

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: https://www.youtube.com/watch?v=OzkcB1lkgGU Hackteria: https://hackteria.org/wiki/DIY_NanoDrop Webcam: https://www.amazon.ca/AUSDOM-1920x1080P-Noise-cancelling-Recording-Computer/dp/B017SRHRR6/ref=sr_1_7?crid=8KPDXFH5IJ93&keywords=hd+webcam&qid=1547857754&sprefix=hd+web%2Caps%2C175&sr=8-7 _________________________________________________________________ Support the show and future projects: Patreon: https://www.patreon.com/thethoughtemporium Store: https://teespring.com/stores/the-thought-emporium Become a member: https://www.youtube.com/channel/UCV5vCi3jPJdURZwAOO_FNfQ/join _________________________________________________________________ My Social Media Pages: Instagram: https://www.instagram.com/thethoughtemporium/ Facebook: https://www.facebook.com/thethoughtemporium/ Twitter: https://www.twitter.com/TTEchironex Website: http://thethoughtemporium.com/ _________________________________________________________________ As always, thanks to my awesome Patrons for helping to make these video possible. Special thanks to: Afrotechmods Alex Alex Vanderpot Alexander Halbleib Alexandre Guhur Andrew Gray Anita Fowler Austin Morris Ben Krasnow Ben Reay Bennet Huch besenyeim Cataract Bumblesnatch Christian Bastos Colby Callahan Danny Chan Daniel Gunn David Golouski Ezekiel Dohmen Elliot Turner Frank Harlad Havana Woody Jase Smith John De Witt Jim Mussared Jonas Abreu Jonathan Anderson Justin Hendryx Kevin Kevin Comerford Kevin Forsythe kn0tsin Kush Agarwal Larry Leon Schutte Matt KI7ZTH Michael Chatzidakis narvutar1324 Nico Schlüter Nicolas Pourtalé Noah Pedersen OmuN Patrick Sweetman Patryk Wycech Paul Emmerich Paul Richmond Peter Scott Philipp Weber Pietro Saccardi Rauni Robert S. Ryan Harvey Simon Hallam Skave Rat Tom Bullock tain TheQuickestBrownFox Vasilis Whence __________ Channel Members: Felix Nica L Weiner Kush Agarwal Anthony M Hunter Bagby

Comments

Leslie Rohde

Justin, that's a great build, and I'm working on improving it specifically for DNA. It start though, it would be great to have a link to the original plans which I did not find using that Google thing. Either way, my improvement is to use a pair of UVC emitters at the key wavelengths required for DNA quantification. At $30 a pop they are not cheap, but should "plug and play" with your design you've already proven. One open question is calibration. Everything I can find references absorption per centimeter, so I'm not sure how the math on a nano-drop spec even works.

thethoughtemporium
>>23140212

I'm actually going a different route for DNA quant. I'm seeing if I can cram all the spec optics into one of the cheap DSLR-microscope adapters. Then I can just take a long exposure, and I've already demonstrated that the laser can be filtered out. Will need to do some calibration, but it's my bet for an easy DNA quantification method. But if your method works let me know. sounds promising potentially. not sure what you mean about calibration? Once you know you are seeing DNA, just make a dilution series from known stock and see what light levels you get. Done. Don't worry about the math, just build a calibration curve and make sure to do everything the same each time

Leslie Rohde
>>23140212

Re: Your new approach. Very interesting. Please tease my mad-Googling skills with some buzz words for what you'll stack into that adapter? Re: My calibration question... purity and concentration are independent factors and the cuvette style devices do the concentration math (in part) on path length through the s ample, but as I was explaining (justifying?) my question, I realized the calibration for a nano-drop is identical. Sure, the cuvette is a fixed size whereas the nano-drop is (more or less) an uncontrolled thickness, but you have to do a water-only baseline on each test anyway. Duh. Size, in this *one* case, really does not matter. That said, building a calibration curve is tricky unless you use LOTS of samples because "the absorbance curve at 280 nm is quite steeply sloped". The technical note doesn't show a graph, but it's bio so I think we can assume it is non-linear? :-) Do I want to characterize that curve imperically with (expensive) samples? And how? Concentration is not the variable, it's purity, and that's orthogonal. Not sayin' no, just thinking. re: Your plan for long exposure. Is it not true that UV degrades DNA? Hence blue light gel-docs? Of course, you need long exposure because you have limited intensity at th e wavelengths of interest, and also therefore, limited impact on the DNA. CA *classic* engineering problem! Which happens faster: DNA damage or development of a usable image. Rather than long exposure, I'm looking to go the other direction: sensitive detector and "flashed" UV lasers at the key wavelengths of 230, 260, and 280. Grab three separate (rapid as possible) exposures and do the ratios in software. Likely more than 100 bucks in lasers, but still a steal compared to commercial, right? Which works better? I sure as hell have not clue (I'm a software/electrical meddling in bio!) but I'll bet that both DNA and image sensors are both highly non-linear in very different ways so it is only by testing we'll know anything at all. Thanks for your feedback! This project is stacked with several others - I'm sure you have no idea what I mean, cough cough - but it is "committed" so I'll keep you posed as details materialize. By all means, please comment further when you have other ideas.

thethoughtemporium
>>23140212

That's a different problem. I personally don't care about purity, just concentration because that's all that matters in my applications. That's why I stain mine with a DNA binding dye so the only thing that glows is DNA and everything else is ignored. To get a proper spectrum and 260/280 ratio to make sure there's no proteins is much harder

thethoughtemporium
>>23140212

Also UV takes a while to damage DNA especially the 405nm I use. Again this is the benifit of the dye. It'll also absorb much of the UV and protect the DNA a bit. That said, another viewer is also sending me some proper deep uv leds and filters to match that he had as overstock at his company so I'll try both methods.

Leslie Rohde
>>23140212

Hmm. Purity is not a problem because you are working on highly robust organisms?

thethoughtemporium
>>23140212

Ecoli, and yeast, ya. I'm not doing mammalian or something where endotoxins are a problem really. And if it was, I'd just find an endotoxin removal kit that works and just always use it if I'm going from bacteria to mammalian as it'll always be contaminated.

Leslie Rohde
>>23140212

It may be that 405 doesn't noticeably damage DNA at all, or is blocked by the dye (although, I am trying unsuccessfully to make a picture of how the dye "gets in the way"). In either case, there are so many variables, many more than we know I think, that testing will out, so I am especially interested in this line of inquiry. Gotta go for now, but I'll post to this thread the UVC emitters I have selected.