Thank you! I am a researcher who works with polarizing microscopes and I am trying to get into fluorescence work. Out of a dozen video lectures on the topic, several textbook chapters, and a pile of research papers, this video is the first time that most of this has made sense. I especially appreciate the detailed description of the filters, most lectures leave a LOT out on those. I still have some advanced questions about the light source, and how UV fits into this (is it also produced by the Hg lamp?), but I finally feel that I am making progress on this!
Thank you for the Lecture Sir,very detailed, I have been working in Diamonds since 1998, I have to study more in Fluorescence of Diamonds, Please provide if you have any information ,
Excellent video, would be nice to see a tutorial on specimens preparation and various staining techniques. I have found specimen preparation and technique is more important than the mechanical understanding of the process. I am sure your techniques would be another excellent tutorial from you! Thanks
This video help me a lot, I use alpha-tubiline antibody, a lot of time to see, reveal microtubules in the cell, but the image (spindle) was not clear, after watching these videos, I did and it works. Thank you.
Thank you for this helpful introduction. I believe the quantum efficiency (QE) explained in this video is more commonly referred to as quantum yield. QE is a term used for the camera or detector that is capturing the photons.
Thank you for the clear and thorough presentation. Is fluorescence microscopy in general really quantitative, though? The local environment of the fluorophore can have an effect on the fluorescence quantum yield and the fluorescence spectrum of the fluorophore, making it difficult to draw a direct correlation between fluorescence intensity and fluorophore concentration. Am I right? I guess this problem can be circumvented by using fluorescence lifetime imaging microscopy, in which the information (i.e., the fluorescence lifetime at each pixel) is independent on fluorophore concentration. Also, no energy is lost during internal conversion, since internal conversion is an isoenergetic process. The excitation energy is lost during the vibrational relaxation process that takes place after the internal conversion process.
Hello, excellent video, I need help, I would like to know which ones, with the advantages and disadvantages of fluorescence, sorry if you do not understand the question well, I speak Spanish.
Is there a microscope which allows to collect all these types of exposures at once for a detailed (colored) video compilation of samples in motion? Rather than via a turret limited to one exposure at a time?
Thank you for the education on fluorescence microscopy. I need to capture a live video of bacteria reaction/dying in presence of house hold disinfectant such as Clorox and Lysol drops. What equipment is best for capturing the video? Would a LED based Dark-field Microscope be good enough or should I consider buying a Fluorescence Microscope and if so what is a good cheaper brand I should look at?
12:00 I'm a bit confused by the diagram. Is S0,0 and S0,2 the same energy or is S0,2 higher? Because it looks like there's a photon of the same wavelength being emitted when an electron drops from S1 to S0,0 and S0,2 but that can't be if they're different energy levels, right?
i have a question. Actually which electron will go upper state. Here the higher energy state of which molecule? If single electron go up, then sometime spin change, why?
hi there, I would like to know about the protein in jelly fish that you can inject into organelles, how does that work. do you have any video you can upload please? thanks
great video tutorial for a starter. Explains really well and elaborated. I appreciate. One thing I need to know is about the bandwidth concept. What is the narrowest bandwidth we can use? SO, is it good to use as narrow range as possible?
Harpreet Kaur hey very good question, it is not about how narrow the bandwidth is, because you need to get as much of the emitted photons as possible (that is the 2nd peak in the graph). Therefore you use a bandwidth that covers as much (wider) of the emitted as possible, without covering the exciting light that you put initially (the first peak in the graph). so to keep it short look at the graph he has in 22:00 and see how the bandwidth is covering as much of the emitted as possible and as little of the exciting as possible. hope that helps
this is really helpful. But please help me how i can adjust emission and excitemnt wavelength 490 to 550nm. And how can i adjust the contrast to make the background totally dark. Please help!! I dont know anything about fluorescent microscope and really i am in need!!!!
You can analyze colocalization in fluorescence microscopy images on iPad using CoLocalizer app, free from App Store: itunes.apple.com/app/colocalizer-for-ipad/id1116017542?l=en&mt
You can see in his eyes all the time he has dedicated in his life to this subject he loves so much.
could not of asked for a better, well presented explanation of fluorescence. Very nicely done.
Thank you! I am a researcher who works with polarizing microscopes and I am trying to get into fluorescence work. Out of a dozen video lectures on the topic, several textbook chapters, and a pile of research papers, this video is the first time that most of this has made sense. I especially appreciate the detailed description of the filters, most lectures leave a LOT out on those. I still have some advanced questions about the light source, and how UV fits into this (is it also produced by the Hg lamp?), but I finally feel that I am making progress on this!
Thank you for the Lecture Sir,very detailed, I have been working in Diamonds since 1998, I have to study more in Fluorescence of Diamonds, Please provide if you have any information ,
Excellent video, would be nice to see a tutorial on specimens preparation and various staining techniques. I have found specimen preparation and technique is more important than the mechanical understanding of the process. I am sure your techniques would be another excellent tutorial from you! Thanks
Thank you so much..no one has explained as good as you. Thank you iBiology🙏
im korean, and i'd appreciate this video that is so intutive and well explained
Thank you so much for this presentation, for the very first time I actually get it!
haha play 32:19, he says fkd up instead of fluorescent :P Great lecture btw
there should be some beeep sound
I went straight to it
i am so impressed how did you even notice that:) great attention, love that for you
Your way of explanation by using tools is really aid in learning....thank you for making such nice and helpful video !!!!
Such a wonderful, in detail presentation. Thank you Sir
It's a very complicated subject, and you are doing a good job here. Thanks
Amazing lecture very simplified but of high level. thank you very much it was very helpful.
This video help me a lot, I use alpha-tubiline antibody, a lot of time to see, reveal microtubules in the cell, but the image (spindle) was not clear, after watching these videos, I did and it works. Thank you.
I have my first course in nanoengineering, this really helped, thanks a lot!
Thank you for this helpful introduction. I believe the quantum efficiency (QE) explained in this video is more commonly referred to as quantum yield. QE is a term used for the camera or detector that is capturing the photons.
Thank you for the clear and thorough presentation. Is fluorescence microscopy in general really quantitative, though? The local environment of the fluorophore can have an effect on the fluorescence quantum yield and the fluorescence spectrum of the fluorophore, making it difficult to draw a direct correlation between fluorescence intensity and fluorophore concentration. Am I right? I guess this problem can be circumvented by using fluorescence lifetime imaging microscopy, in which the information (i.e., the fluorescence lifetime at each pixel) is independent on fluorophore concentration. Also, no energy is lost during internal conversion, since internal conversion is an isoenergetic process. The excitation energy is lost during the vibrational relaxation process that takes place after the internal conversion process.
Hello, excellent video, I need help, I would like to know which ones, with the advantages and disadvantages of fluorescence, sorry if you do not understand the question well, I speak Spanish.
thank you so much - really helped me get into my term paper about fluorescence microscopy
Thank you for knowledge. 😊❤
Is there a microscope which allows to collect all these types of exposures at once for a detailed (colored) video compilation of samples in motion? Rather than via a turret limited to one exposure at a time?
Thank you for the education on fluorescence microscopy. I need to capture a live video of bacteria reaction/dying in presence of house hold disinfectant such as Clorox and Lysol drops. What equipment is best for capturing the video? Would a LED based Dark-field Microscope be good enough or should I consider buying a Fluorescence Microscope and if so what is a good cheaper brand I should look at?
at 16:02, he says that there's an intereference of light. what causes the phase difference for interference to occur ?
Excellent lecture and demonstration. Really impressive.
12:00 I'm a bit confused by the diagram. Is S0,0 and S0,2 the same energy or is S0,2 higher? Because it looks like there's a photon of the same wavelength being emitted when an electron drops from S1 to S0,0 and S0,2 but that can't be if they're different energy levels, right?
i have a question. Actually which electron will go upper state. Here the higher energy state of which molecule? If single electron go up, then sometime spin change, why?
Great, clear, and informative presentation. Thank you
frodo teaching me biology, gotta love it
Great video, do you know how to clean the oil inside of the objective? Thanks.
very well explained,video makes clear the principle and working of Fluorescence microscopy,thanks
Great job, well explained, thak you for everything
These videos are amazing. Thank you
Good job. This is a very thorough lecture.
You cannot really claim FM to be quantitative, because in most cases what you are imaging is the fluorescent labels you have added to your sample.
great explanation! thank you!
hi there, I would like to know about the protein in jelly fish that you can inject into organelles, how does that work. do you have any video you can upload please? thanks
love it! At last I understand how those "cubes" really work!
Perfect video!!! Thanks so much
Does immunofluorescence microscopy emits x-ray? Or gamma rays? How much immunofluorescence microscop safe??
good video!
p.s did you have a haircut between the scenes? :D
what is the name of the multiple cube holder?
great video tutorial for a starter. Explains really well and elaborated. I appreciate. One thing I need to know is about the bandwidth concept. What is the narrowest bandwidth we can use? SO, is it good to use as narrow range as possible?
Harpreet Kaur hey very good question, it is not about how narrow the bandwidth is, because you need to get as much of the emitted photons as possible (that is the 2nd peak in the graph). Therefore you use a bandwidth that covers as much (wider) of the emitted as possible, without covering the exciting light that you put initially (the first peak in the graph). so to keep it short look at the graph he has in 22:00 and see how the bandwidth is covering as much of the emitted as possible and as little of the exciting as possible.
hope that helps
yes. It makes sense. Thanks
can you explain a negative stoke shift?
that was great, i like it so much, it helps so much... thank so much for all of this great demonstration and information thaaaaaaaaank u sir
this is really helpful. But please help me how i can adjust emission and excitemnt wavelength 490 to 550nm. And how can i adjust the contrast to make the background totally dark. Please help!! I dont know anything about fluorescent microscope and really i am in need!!!!
Thank you! Very clear lecture.
Really awesome explanation Sir
Gratefully, thank you ❤❤❤
Very well explained, thank you so much!
Thank you so much! you explained really well.
Perfect presentation, thank you very much
what is differance between
Fluorescence imaging and florescence microscopy
microscopy is a part of imaging ! bioimaging is a wide approach for diagnosis and molecular studies, which includes microscopy, x ray imaging etc etc
Great presentation, thank you!
Really, really great! So helpful!
why dying inner cell and extracellular ?
Thank you for this video! Very helpful
excellent video!! thank you!
Muchas gracias, muy útil, muy amable, muy bien explicado
Incredible ! Than you
Great video!
Love it, thanks Nico! :)
> Thank You so much for uploading it
Well presented, thank you
Excellent!
Thank you
Amazing!!!
Thank you very much
thanks for your video
Thank you for nice video.
Thanks.. awesome video!
thanks for the video!
good one
bbbeautiful sir
that's my uncle :)
You can analyze colocalization in fluorescence microscopy images on iPad using CoLocalizer app, free from App Store: itunes.apple.com/app/colocalizer-for-ipad/id1116017542?l=en&mt
很棒啊
👍🏻♥️
Rutger Hauer between acting jobs.
helpfulvideo
good
unsharp images for much money
Fluorescence die during experiment ;P
IGNOU BSCBCH 😁😊🚩