Fluorescence activated cell sorting (FACS)

[@anythinggoes4588]

I'm yet to meet a person on this planet, the milky way galaxy, that's completed their degree without Shomu! :) Thank you!

[@shomusbiologyofficial]

+Anything Goes glad I can help

[@comoplaysdestiny5106]

im 30 seconds in and already know this is gonna be super helpful for my technique write up paper in developmental biology. thanks!!

[@ipeksolmaz4643]

incredible teaching ability

[@shomusbiologyofficial]

Thank you

[@mayukhroy5393]

This video was too helpful Sir.

[@shomusbiologyofficial]

You're welcome

[@ArchismanGanguly]

You can make topic understand for everyone 😉

[@shomusbiologyofficial]

You're welcome

[@carolinegronnier6888]

Thank you so much for sharing this really clear video

[@shomusbiologyofficial]

You're welcome

[@ravianandrao7761]

Thank you so much shomu bro...

[@kusfierce4328]

Sir ..you r always ..a saviour ... Love u sir .. u make the topic soo easy to get understand ... ❤️❤️ Love from heart

[@shomusbiologyofficial]

Thank you so much for appreciating my efforts

[@franciscoperez6132]

Excellent explanation, salam amelicum❤👍

[@shomusbiologyofficial]

Thank you

[@isabellajung9834]

Thank you very much for this good explanation! I have one question: Where exactly is the difference between die amount of proteins and the number of cells expressing this protein. Is this about how much of this proteins are on the cells, so more about the quality?

[@sushreesangitapriyadarshin6441]

This is how the cells get charged... The flow chamber, and the flow stream, is charged (± v where v typically lies between 50 and 150 V) at the moment a cell of interest is inside the droplet . The stream of droplets passes through a pair of charged plates (± 5000 V) so that the charged droplets are deflected and collected together with the cell container

[@The9988111309]

If a potential difference is applied on the flow chamber containing all sorts of cells then the whole stream of droplets will be charged right? Why only the cell of interest? The more complex way is that the cell of interest is charged based on the scattering where the pattern is detected and when the flow stream exits forming droplets at the bottom then they are charged accordingly with an electric ring present, then they are separated with the help of charged plates in the end.

[@rezafarahmand2695]

Thank you,good for you👏👏👏

[@shomusbiologyofficial]

You're welcome

[@vgreddy_velma]

very useful for beginners

[@shomusbiologyofficial]

You're welcome

[@TheSebastiandiego]

Just to clarify his explanation is wrong. First the cells that have the florescence will get charged with an electrical charging ring, they are then differentiated by electrostatic deflection. they are not naturally charge they acquire a charge to differentiate the ones that do and don't have the flourecent antibodies

[@igurujiaadharinstitute]

you must know about Mr. hemant ....he is CEO OF a leading startup FLOW Jo in middle east and south east asia.....EXPERT of FLOW cytometry........MSc LIFE science fron JNU,.......PhD from max plank germany......Post Doc from chicago university..........

[@subhamkarmakar3320]

You are right

[@PureCool-k2o]

This comment may have been from 7 years ago.. but I have a doubt, if cells that emit fluorescence are negatively charged (and hence deflected ) then only the anode side and uncharged side of the cell sorter collects cells right? Is there any cases where cells acquire positive charge to get deflected to the cathode and collect there?

[@kalpanajaiswar5025]

Thank you so much sir 🙏🙂

[@shomusbiologyofficial]

You're welcome

[@risharajkhowa5060]

thank u,so much Sir

[@shomusbiologyofficial]

You're welcome

[@muhammadahmad-ft7fv]

awesome job bro ,,,keep it up ..ALLAH bless u

[@ullaskotihal1911]

Thanks bhai Fan from Karnataka 😍 We wanna meet you

[@shomusbiologyofficial]

You're welcome. Glad to hear that you are getting benefit from the videos

[@ullaskotihal1911]

Where are you from bhai?

[@smarakipattnaik685]

Sir, how many cells can be sorted by charging method in this technique ?

[@gubeshgunaratnam4615]

very nice explanation

[@ankitalankar6042]

thank you sir

[@hemantagrawal3772]

My dear there is no use of radioactivity in flow cytometry. Please correct your sentence. We use fluorescent proteins or dyes but no radioactive material

[@pranjaldhaka435]

+Hemant Agrawal so FACs just record the fluorescence of the tags or flourochromes attached to the cells right?

[@hemantagrawal3772]

+Pranjal Dhaka In flow cytometry, cells or cell like particles can be analyzed based upon their physical properties like cell size and cell complexity plus cells of interest can also be identified in a mixture by tagging its specific markers (surface/intracellular) by antibodies labeled with fluorescent tag. Sometimes cells can also be studied using fluorescent dyes without any antibodies for example viability assay, side population determination etc. Please see a link, where flow cytometry is explained in a very nice way www.d.umn.edu/~biomed/flowcytometry/introflowcytometry.pdf

[@igurujiaadharinstitute]

PRANJAL DHAKA you must know about Mr. hemant ....he is CEO OF a leading startup FLOW Jo in middle east and south east asia.....EXPERT of FLOW cytometry........MSc LIFE science fron JNU,.......PhD from max plank germany......Post Doc from chicago university..........

[@Captainlab01]

Great to see u Sir

[@DrRemedius]

37 target channels acquired

[@soniyavincent9116]

May i know that is this answer for flow cytometry and fluroscence

[@hosseinghadamyari3513]

very good

[@wonderfulIndia1]

Sir flow cytometry bhi ise hi bolte h kya sir plzz reply

[@shomusbiologyofficial]

Yes

[@sinamt2982]

How do the different cells get the different charges to seperate them in the electric field?

[@vijetasingh416]

Sir please give lacture on the quenching fluorescence and phosphorescence please

[@eggnogs]

Having used the FACS machine for multiple years, I can verify that this information is incorrect. Also fluorescence is not radiation. You need to go back to school and pay attention

[@shibrajpradhan6849]

😂 something is burning

[@pamelajulia1610]

The intensity is on the x axis. Not y

[@love.vibe.official]

GAL4/UAS.... Please ..... Upload.. This topic

[@avinashkumargudadurkalmath7368]

R u with me I m going to xenotransplantation process practically from monkey genes to human tell what are the steps involved in that

[@rijilfreekdaredevil]

Thank u sir

[@shomusbiologyofficial]

Welcome

[@MarceloZerillo]

Hi Suman, I have a question. I am reading a paper in which FACS was used and for a control of the experiment a batch of the cells was treated with 70% ethanol in order to kill them. They report the counting of the dead cells, but I wonder if it is still possible to detect those dead cells using FACS, since the membrane would be disrupted and the cell content would be lost. Any guess? Thank you.

[@pranjaldhaka435]

+Marcelo Zerillo I think the purpose of the control in the experiment was to show the relative inability of the fluorescent tags to attach to the cells which got destroyed due to ethanol treatment. It still depends on the fluorescent tag used though because some tags attach to a specific protein under specific conditions,.

[@hemantagrawal3772]

+Marcelo Zerillo In Flow cytometry, people use viability dyes like PI, 7AAD, DAPI etc to detect dead cells. The principle is that dye gets bind to nucleic acids once the membrane is compromised and we can easily differentiate between dead and live . It has nothing to do with the non-attachment of fluorescent tags to the dead cells. Please see this link for a flow figure www.uvm.edu/medicine/flowcytometry/documents/ViabilityusingPI_000.pdf

[@MarceloZerillo]

+Pranjal Dhaka thank you so much for your answer.

[@MarceloZerillo]

+Hemant Agrawal thank you so much for your answer.

[@surajsaksena]

totally wrong explanation of sorting..

[@shomusbiologyofficial]

+Suraj Saksena really? Please explain your points

[@2lefThumbs]

I explained quite clearly where you were wrong over a year ago, and you deleted my reply, if you leave this intact, I'll gladly reconstruct my critique

[@mercy9386]

@@2lefThumbs UMMM can I please get the correct explanation then please? I need it ASAP

[@atulanand7993]

Wrong.....👎🏿

[@noorsbiology7580]

ur x and y axes are switched I think

[@sajidmicrobiology]

you need to go back to school dear... why are spreading wrong information ... just delete it

[@tothbence1995]

Shomu's Biology, please delete your video because it's quite popular and spreads false information.