Really nice video. It let us know both, things common between ELISA and elispot and also what makes them different from each other. Such animations in your videos are always helpful to remember things in a better way.
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Good point....thanks for the feedback ...I should have noticed it. These feedback would help to improve the quality of the upcoming content. Thanks again
What is the advantage here in comparison to flow cytometry? In flow cytometry we do not look for "shadows" after cells, but we can actually count how many Th17 cells are there and it is much easier and faster than Elispot. Also in Elispots we do have SFUs often and it is hard to say how many cells were really there. I just don't understand why not just using flow cytometry? If someone uses PBMCs they won't know which cell really produced IFN (many do it) so Elispot will show in this case only shade of a cytokine and not the producing cells (we won't know which cell produced IFN in such case). An honest question, because I am new to Elispots and old to flow ;) I found my answer: "Flow cytometry can determine the number of cells that produce IFNγ, but not their secretory activity and the amount secreted by them. ELISPOT quantifies the frequency of IFNγ-secreting cells, but not the number of producing cells nor does it measure the amount of IFNγ secreted from cells."
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Can you please clear a doubt? One cell can produce many molecules of cytokines which can subsequently bind to more than one capture antibody, and hence produce more than one spot. So why is it that one spot equals to one cell?
I also don't understand. Can someone explain this? How do you know that the certain spot is from a different cell. Can be from different amount of a cytokine produced by the same cell.
@@olamarca7171 i think its because, each cell produce lots of cytokines and these cytokines will bind to nearest antibody. The cytokines naturally will diffuse in a small circular radius around that singular cell. Then, in this small circular area around the singular cell, the antibodies binded with cytokine will form complexes with secondary enzyme - and due to their very high concentration of complexes in that small circular radius around the singular cell, we see a single spot. Hence why one spot is from one cell.
So in ELISPOT, no of spots indicates no of cytokine secreting cells, however, I am not satisfied with this theory because I feel that one cell can secrete multiple cytokines, for example the IFN cytokine represents TH1 cell, and since TH1 cells secrete multiple IFN cytokine, wouldn't it be wrong to state that each no of spots indicates no of cytokine secreting cells. please help me clear this doubt
Sir if i want to analyse that which cytokines are release from the particular immune cell in respnse to a particular infectious agent which approach i can apply?
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@@animatedbiologywitharpan I would use elispot to determine if my pbmc T cell culture expansion worked by determining how many/frequency of specific cytokine (for example IL-2) secreting cells, correct?
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Really nice video. It let us know both, things common between ELISA and elispot and also what makes them different from each other. Such animations in your videos are always helpful to remember things in a better way.
U r a blessing to the biology exam aspirants 🙏... Thanks a lot sir
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Very helpful & easy to understand
Very insightful, helpful and nicely done! Thank you for your work, aand I hope you are feeling appreciated.
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Very good animation. U forgot to mention abt stimuli. U need to add stimuli so that it will stimulate the cell to produce cytokines.
Good point....thanks for the feedback ...I should have noticed it. These feedback would help to improve the quality of the upcoming content. Thanks again
@@animatedbiologywitharpan I wonder how u responds to doubts..is it u watch it again or u knew it.
@@rashmiranjansahoo3975 I watched it quickly and saw i did not mention about a stimulus which might induce cytokine secretion.
Brilliant video, extremely helpful
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Serum and plasma are acellular. You need to use PBMCs for this assay.
What is the advantage here in comparison to flow cytometry? In flow cytometry we do not look for "shadows" after cells, but we can actually count how many Th17 cells are there and it is much easier and faster than Elispot. Also in Elispots we do have SFUs often and it is hard to say how many cells were really there. I just don't understand why not just using flow cytometry? If someone uses PBMCs they won't know which cell really produced IFN (many do it) so Elispot will show in this case only shade of a cytokine and not the producing cells (we won't know which cell produced IFN in such case). An honest question, because I am new to Elispots and old to flow ;) I found my answer: "Flow cytometry can determine the number of cells that produce IFNγ, but not their secretory activity and the amount secreted by them. ELISPOT quantifies the frequency of IFNγ-secreting cells, but not the number of producing cells nor does it measure the amount of IFNγ secreted from cells."
The only benefit for this technique is the low cost compared to flowcytometry
very well explained helped me a lot thank u
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Very nice explanation!
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Helpfull👍✨️
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Very Nice video, thank so much 🇩🇿
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Very nice explanation sir ☺☺
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keep on going,we support yoy
You are gem Arpan fantastic
Thanks a lot my friend ❤
Can you please clear a doubt? One cell can produce many molecules of cytokines which can subsequently bind to more than one capture antibody, and hence produce more than one spot. So why is it that one spot equals to one cell?
Good question....email me your contact we can chat in WhatsApp...too much to write in a comment
@@animatedbiologywitharpan your email?
I also don't understand. Can someone explain this? How do you know that the certain spot is from a different cell. Can be from different amount of a cytokine produced by the same cell.
@@olamarca7171 i think its because, each cell produce lots of cytokines and these cytokines will bind to nearest antibody. The cytokines naturally will diffuse in a small circular radius around that singular cell. Then, in this small circular area around the singular cell, the antibodies binded with cytokine will form complexes with secondary enzyme - and due to their very high concentration of complexes in that small circular radius around the singular cell, we see a single spot. Hence why one spot is from one cell.
Plz send some research area for elispot cytokine secreting for therapeutics and vaccine
So in ELISPOT, no of spots indicates no of cytokine secreting cells, however, I am not satisfied with this theory because I feel that one cell can secrete multiple cytokines, for example the IFN cytokine represents TH1 cell, and since TH1 cells secrete multiple IFN cytokine, wouldn't it be wrong to state that each no of spots indicates no of cytokine secreting cells. please help me clear this doubt
very informative
Sir if i want to analyse that which cytokines are release from the particular immune cell in respnse to a particular infectious agent which approach i can apply?
Thanks! Will done!
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GOD bless. Too satisfied
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🙏
Sir....could u plz clear my doubt
ELISPOT is used for measuring cytokines or the secretory cells like T-cells??
jabi fatma cytokines secreting cells are detected using this assay.....for cytokines only ELISA is enough
Thanku so much
@@animatedbiologywitharpan I would use elispot to determine if my pbmc T cell culture expansion worked by determining how many/frequency of specific cytokine (for example IL-2) secreting cells, correct?
@@user-do2iq8rc5q yes you can use it for this purpose
Sir in which diagnostic center this test is avaliable sir
That depends .....generally these center's deals with infection and immune systems...its there in NII Delhi
Love ya
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Ah, Biorender :)
Unfortunately I do not understand the English from Indian people
Thanks for the feedback, will try to improve in future
jiayous
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you are reading not explaining brother we are getting clarity on the topic 🤨
I didn't get your point....please elaborate