This video is great because you smile & sound enthusiastic, & because the video is visually interesting, well organized, technically accurate & funny. You also use relatable examples. I loved the "What?!!!" parts. Nice job!
Im preparing for a biology content exam to teach biology in high school and never in all my years of high school and college did I even hear the word Spectophotometry, so THANK YOU. This helps so much!
@@YouTooBio Can I ask what if I want to measure the absorbance of methylene blue after I put carbon coated sand with it ... If I press ZERO-BLANK in the device it gives me (insert empty cuvette or distilled water)... should I do that or insert a cuvette contains MB from the stock solution ??
@@bxwn8931 1) use the same concentration of MB for the blank, 2) the spec only works for dissolved solutions - if the sand sinks to the bottom you won't get a meaningful measurement (unless the sand changes the color of the MB, in which case mix them for some amount of time, then measure just the liquid portion with the spec)
Omg,i don't have words to express my gratitude to you,you explained the things so well,it was my 4 or 5 video on the same topic but u r the only one who made me clear
On what base do you choose a certain wavelength to measure at ? For example if the sample color is red you choose the wave length of red color? Thanks a lot
Great question! If you don't have a protocol telling you what wavelength to use, then you need to determine the absorption spectrum for your sample - do this by measuring the absorbance (A) across the visible spectrum (~400-700nm), measuring every 10 or 25 nm or so. This will give you a curve showing you the wavelength with the highest amount of absorbance - that's the wavelength you'll use for subsequent experiments testing concentration of that substance. For your second question, if your sample is red, then your sample won't absorb red light - it reflects red light, and that's why it looks red! So, if you use red light (wavelengths 600-700nm) for a red sample, you'll have an absorbance near or at 0! For further reading: chem.libretexts.org/Bookshelves/Physical_and_Theoretical_Chemistry_Textbook_Maps/Supplemental_Modules_(Physical_and_Theoretical_Chemistry)/Kinetics/02%3A_Reaction_Rates/2.01%3A_Experimental_Determination_of_Kinetics/2.1.05%3A_Spectrophotometry
@@YouTooBio Ok great thanks. Last question 🙏🏻 why do we measure at certain wave length? We can get the conc. Of the sample from standard curve. But if we didn't measure at certain wavelength it will give us zero or negative. Why? I googled for the answer a lot but i didn't find a satisfying answer. Second, I heard that If we want to measure wavelength for a specific color we use the color that's before it or the color that completes it I think😅 Is that true?
It's a lot easier to measure at one set wavelength than to do a standard curve for each sample! Generally it's true that if your sample is one color, you will use the opposite color (on the color wheel) for absorbance, which has to do with why colors look the way they do. But it's hard to predict for some samples that don't have obvious color "opposites" - like a culture of bacteria. That's why you do the absorbance curve to determine the optimal wavelength to use.
hi, so is the readout of absorbance is the OD600 value directly? or do i need to calculate the OD based on the absorbance using some formulas? thank you!
This may sound like a dumb question, but do I have to put a blank every time before I put a sample cuvette into the spectrophotometer? Or is the blank's absorbance already recorded once you set it up?
Thank you for this excellent video! Would you mind sharing the make/model of that particular spectrophotometer? And also, any suggestions for a good spectrophotometer that could be used at home? (Less than $5k would be ideal :) I'm researching various components of sunscreen but I don't have access to a professional lab!
Hi Kevin! The spectrophotometer in this video is a Thermo Scientific Genesys 10UV. There are some on eBay. Also, check out Sebastian Cocioba at twitter.com/ATinyGreenCell who is doing lots of biology in a DIY home lab and eagerly helps other DIY scientists!
Can I ask what if I want to measure the absorbance of methylene blue after I put carbon coated sand with it ... If I press ZERO-BLANK in the device it gives me (insert empty cuvette or distilled water)... should I do that or insert a cuvette contains MB from the stock solution ??
Hello, I'm medical student. I need some explanation about the difference between UV-Vis spectrophotometer and Nanodrop spectrophotometer. What is the advantage of both spectrophotometer? Please help me, thank you 🙏
Great job ! Never thought it can be this interesting. I have a question tho. Could using the spectrometer for multiple times say over 20 with only using the blank reagent just onec at the begining alter the results for final measurements? Im in a bio lab in college. And there are over 50 students who use the device before me -cuz im down the list - and the blank reagent is used just once at the begining. Anyhow i always get the results wrong 3 out of 4 times . So i've been thinking could it be the device ? Or do i just suck at using the micropipette?
It depends on 1) the device, does it drift? Probably not but I've seen old specs have unreliable results, so maybe? 2) whether your sample settles between preparation and measurement. The best way to check is to measure your sample as usual, re-blank, then measure your sample again. If you get two different results, then yes it's a problem to blank once before all the students measure their samples. See if your instructor will let you try that.
Ma’am, what if I set the wavelength at 340 nm and set the meter for 100%T, then I noticed that the 100%T meter indication is not stable. What seems to be the problem and what should I do?
That is outside my expertise, sorry! Maybe you can find your answer here? www.chem.uci.edu/~dmitryf/manuals/Fundamentals/Fluorescence%20Spectroscopy.pdf
@@YouTooBio sulphuric acid, water and dissolved wood particles.. i did klason lignin treatment on empty fruit bunches and I wanted to find out the absorbance of my samples after the treatment
Hi my Teacher I am a student phD in Environmental Engineering, Iraq. l need to measure bacteria concrete in media broth, by spectrometer to complete my research..can I get email for you, please.
Thank you for this excellent video! Would you mind sharing the make/model of that particular spectrophotometer? And also, any suggestions for a good spectrophotometer that could be used at home? (Less than $5k would be ideal :) I'm researching various components of sunscreen but I don't have access to a professional lab!
Sorry I don't know the make and model of the spectrophotometer in my video! As a scientist working without lab access, I suggest you check out Sebastian Cocioba on Twitter - he does DIY biology and would likely have lots of advice twitter.com/atinygreencell
Spectrovis plus is the cheapest but accuracy not too good Personally I recommend ocean insight I have STS - Vis as Edu kit with all light source, cuvette holder They used to sell as Edu Kit which is much cheaper for all components But now STS series replaced by ST spectrometer , STS and ST are same size Those ST/STS are small but they rival the big one like Shimadzu, Agilent , Thermo fisher (the one in video) in terms of accuracy of absorbance
This video is great because you smile & sound enthusiastic, & because the video is visually interesting, well organized, technically accurate & funny. You also use relatable examples. I loved the "What?!!!" parts. Nice job!
This is fun to watch! I barely see informative/educational videos like these, pls make more!
Thanks - so glad to hear that it's informative AND fun! I'm definitely making more :)
the way i understand you more better than my laboratory prof 😭😭 thank you for this
Im preparing for a biology content exam to teach biology in high school and never in all my years of high school and college did I even hear the word Spectophotometry, so THANK YOU. This helps so much!
Yay glad it was helpful!
Thank you for helping me to understand what this machine does. You did it in plain english !
I love this chanel. It explains everything simply and to the point. I dont even need to know much English to understand. Great job!
Sounds like the coolest teacher ever..thanks from Jordan
I really appreciate your videos Jamie! Very well planned out and super comprehensive! You keep making 'em and I'll keep watching 'em!
Thanks!! Glad the video was helpful
@@YouTooBio Can I ask what if I want to measure the absorbance of methylene blue after I put carbon coated sand with it ... If I press ZERO-BLANK in the device it gives me (insert empty cuvette or distilled water)... should I do that or insert a cuvette contains MB from the stock solution ??
@@bxwn8931 1) use the same concentration of MB for the blank, 2) the spec only works for dissolved solutions - if the sand sinks to the bottom you won't get a meaningful measurement (unless the sand changes the color of the MB, in which case mix them for some amount of time, then measure just the liquid portion with the spec)
I love your Bill Nye vibe -- making science fun and engaging!
How come this does not have more viewers and suscribers? this is great!!!
I love you. You just saved my exam
Wow! I really enjoyed watching the whole video.
Your way of explaining is very engaging! Thanks!!!
These is amazing.. please make more videos like these on hands on techniques required in laboratory
This is fantastic! You're really enthusiastic in your videos and they're fun to watch. I love the "What?!" sections!
Thanks! Glad you enjoyed them!
great video, about to use one of these for my gymnasium exam (=
Very fun to watch! Thanks 💗💗watching this as a prep for the spectrophotometer bio lab🌚
Omg,i don't have words to express my gratitude to you,you explained the things so well,it was my 4 or 5 video on the same topic but u r the only one who made me clear
This video was so helpful and entertaining! Thank you so much.
Yay glad it was helpful!
What a great and simple explenation, thank you very much!
Such a nice and easy video to watch, great job!
Thanks so much!
beautifully explained and demonstrated.. THANK YOU
Glad it was helpful!
Thank you very much! It's so useful for our online class❤️
Glad it was helpful!
Another winner, Jayme!
On what base do you choose a certain wavelength to measure at ?
For example if the sample color is red you choose the wave length of red color?
Thanks a lot
Great question! If you don't have a protocol telling you what wavelength to use, then you need to determine the absorption spectrum for your sample - do this by measuring the absorbance (A) across the visible spectrum (~400-700nm), measuring every 10 or 25 nm or so. This will give you a curve showing you the wavelength with the highest amount of absorbance - that's the wavelength you'll use for subsequent experiments testing concentration of that substance.
For your second question, if your sample is red, then your sample won't absorb red light - it reflects red light, and that's why it looks red! So, if you use red light (wavelengths 600-700nm) for a red sample, you'll have an absorbance near or at 0!
For further reading: chem.libretexts.org/Bookshelves/Physical_and_Theoretical_Chemistry_Textbook_Maps/Supplemental_Modules_(Physical_and_Theoretical_Chemistry)/Kinetics/02%3A_Reaction_Rates/2.01%3A_Experimental_Determination_of_Kinetics/2.1.05%3A_Spectrophotometry
@@YouTooBio
Ok great thanks.
Last question 🙏🏻
why do we measure at certain wave length?
We can get the conc. Of the sample from standard curve. But if we didn't measure at certain wavelength it will give us zero or negative. Why?
I googled for the answer a lot but i didn't find a satisfying answer.
Second, I heard that If we want to measure wavelength for a specific color we use the color that's before it or the color that completes it I think😅
Is that true?
It's a lot easier to measure at one set wavelength than to do a standard curve for each sample! Generally it's true that if your sample is one color, you will use the opposite color (on the color wheel) for absorbance, which has to do with why colors look the way they do. But it's hard to predict for some samples that don't have obvious color "opposites" - like a culture of bacteria. That's why you do the absorbance curve to determine the optimal wavelength to use.
hi, so is the readout of absorbance is the OD600 value directly? or do i need to calculate the OD based on the absorbance using some formulas? thank you!
This may sound like a dumb question, but do I have to put a blank every time before I put a sample cuvette into the spectrophotometer? Or is the blank's absorbance already recorded once you set it up?
It's already recorded once you set it up, so you don't have to set it every time. See 3:57
I love this how you explaining this stuff. Thank you.
Thank you for this excellent video! Would you mind sharing the make/model of that particular spectrophotometer? And also, any suggestions for a good spectrophotometer that could be used at home? (Less than $5k would be ideal :) I'm researching various components of sunscreen but I don't have access to a professional lab!
Hi Kevin! The spectrophotometer in this video is a Thermo Scientific Genesys 10UV. There are some on eBay. Also, check out Sebastian Cocioba at twitter.com/ATinyGreenCell who is doing lots of biology in a DIY home lab and eagerly helps other DIY scientists!
@@YouTooBio Awesome, thank you!
Well explained. Keep doing this.
nice teaching ,can you teach me about soil fertility analysis by spectrophotometry?
I need your help in understanding something about preparing my solution in the cuvette
Very useful
Thanks maam so much for this informative video
Please make more such amazing videos...
how can we take digital photo of a dyed fabric in spectrophotometer?
Thank you very much, very helpful video
I'm really Thankful for u ...God bless u
This video is great! Informative and fun.
Thanks!
Can I ask what if I want to measure the absorbance of methylene blue after I put carbon coated sand with it ... If I press ZERO-BLANK in the device it gives me (insert empty cuvette or distilled water)... should I do that or insert a cuvette contains MB from the stock solution ??
Thanks, it's easy to understand
Yay! Glad it was helpful
it was amazing video thanks
do u know where I can get an accurate but cheap spectrophotometer?
Hello, I'm medical student. I need some explanation about the difference between UV-Vis spectrophotometer and Nanodrop spectrophotometer. What is the advantage of both spectrophotometer? Please help me, thank you 🙏
The difference is in the volume needed to measure the sample. Nanodrop needs only a couple microliters
This was AWESOME, thank you so much :)))
Great job !
Never thought it can be this interesting.
I have a question tho. Could using the spectrometer for multiple times say over 20 with only using the blank reagent just onec at the begining alter the results for final measurements?
Im in a bio lab in college. And there are over 50 students who use the device before me -cuz im down the list - and the blank reagent is used just once at the begining. Anyhow i always get the results wrong 3 out of 4 times .
So i've been thinking could it be the device ? Or do i just suck at using the micropipette?
It depends on 1) the device, does it drift? Probably not but I've seen old specs have unreliable results, so maybe? 2) whether your sample settles between preparation and measurement. The best way to check is to measure your sample as usual, re-blank, then measure your sample again. If you get two different results, then yes it's a problem to blank once before all the students measure their samples. See if your instructor will let you try that.
Great video.
Can it measure the concentration of dye in drinks?
Yes!
Can you show us how to change the nm 🥺
2:40 "Step 1: set the wavelength"
can spectrophotometer detect mineral content of the cells through emitting light through the skin?
Thnanks alot great vid!
great video! thank you for making this!
Glad you liked it!
Good job
very good video thanks! ☺️
Glad you liked it!
very helpful!
Thanks!
Ma’am, what if I set the wavelength at 340 nm and set the meter for 100%T, then I noticed that the 100%T meter indication is not stable. What seems to be the problem and what should I do?
Interesting problem! I don't know! It may depend on your spectrophotometer make and model. Consulting the operating manual would be my next step.
what is the difference of primary filter and differential filter?
I'm not sure - are you talking about fluorescence spectrophotometry?
Yes :(
That is outside my expertise, sorry! Maybe you can find your answer here? www.chem.uci.edu/~dmitryf/manuals/Fundamentals/Fluorescence%20Spectroscopy.pdf
Thanks you very much !!!!
nice video ... thank you :)
Thanks!
some of yall r gonna get mad at me 4 this but im writing a fan fictional work and i had to watch this to write a scene
Wow that's a new one! Share the fan fic when it's ready?
I want to read your fanfic now
amazing video. thanks loved it. haha
Glad you enjoyed it!
Why my reading changes consecutively after the first try
What is your sample? If it has heavy particles, they can settle out of the solution over time, which will change the reading.
@@YouTooBio sulphuric acid, water and dissolved wood particles.. i did klason lignin treatment on empty fruit bunches and I wanted to find out the absorbance of my samples after the treatment
However, I just took the supernatant of the samples
Yeah sounds like large particles. Be sure to shake your sample before pipetting into the cuvette and then take the reading quickly
👏👏👏
nice job explaining Spec but you need to speak slowly. You speak toooo fast. Thank you
Hi my Teacher
I am a student phD in Environmental Engineering, Iraq. l need to measure bacteria concrete in media broth, by spectrometer to complete my research..can I get email for you, please.
poggers video !
hi Jayme you very fast speak😁
who’s here from my biology class watching this 🤡
Buy one and play in home 😂
Thank you for this excellent video! Would you mind sharing the make/model of that particular spectrophotometer? And also, any suggestions for a good spectrophotometer that could be used at home? (Less than $5k would be ideal :) I'm researching various components of sunscreen but I don't have access to a professional lab!
Sorry I don't know the make and model of the spectrophotometer in my video! As a scientist working without lab access, I suggest you check out Sebastian Cocioba on Twitter - he does DIY biology and would likely have lots of advice
twitter.com/atinygreencell
This model is Thermo fisher genesys 10 spectrophotometer
go to 2:40
Spectrovis plus is the cheapest but accuracy not too good
Personally I recommend ocean insight
I have STS - Vis as Edu kit with all light source, cuvette holder
They used to sell as Edu Kit which is much cheaper for all components
But now STS series replaced by ST spectrometer , STS and ST are same size
Those ST/STS are small but they rival the big one like Shimadzu, Agilent , Thermo fisher (the one in video) in terms of accuracy of absorbance