I am still bit confused about charging the drops which we want to sort. We need to charge them at the break-off point but in the picture it shows that the flow stream is charged which means all type of cells will be charged. It makes sense to charge them according to the scattering spectrum observed, in that way we know which to sort or not. The charging is programmed according to scattered pulse values we want. Please do elaborate this otherwise I will pull all my remaining hair out. Regards, Akshit
In a cell sorter, particles are delivered in a buffered salt solution, capable conducting electrical current. We decide (based on the pattern of scatter and fluorescent signals) at the laser interrogation, if we want to sort a particle or not. We have no a bit of time to execute this sort decision, because the particle needs to travel from the laser interrogation to the droplet break off point. Once arrived there, we are charging the whole stream (here you´re right) and the leaving drop is carrying this charge and can be deflected. This works, because electrical current is travelling fast and is arriving at the same time at the break off point as the particle. (PS: Watch the slide around minute 10 “Defining drop delay - Accu Drop”: In the bottom of the screenshot you see numbers with sliders for 2nd, 3rd and 4th drop. This is correcting for potential remaining charges on the stream, once a drop has left. It ensures, that the next drop has a net-charge of zero before it can be charged again for the next sort decision.)
first we decide if we want to sort this particle and in which bucket, then we apply positive or negative charge accordingly to the whole stream, then we wait for the droplet to break off, and then we ground the stream so it loses all charge and becomes neutral again until new sort
![Multicolor flow cytometry - Understanding the basics [WEBINAR]](http://i.ytimg.com/vi/8KQmiQksubY/mqdefault.jpg)
Nice introduction to FACS! Greatly presented, Steffen, thank you!
SIR COULD YOU PLLZ PROVIDE SOME REFFENCE BOOK FOR FACS ??
Loved the video. Thanks for the upload!
Very focused on optimization. (14:14)
3:20 Apparatus.
I am still bit confused about charging the drops which we want to sort. We need to charge them at the break-off point but in the picture it shows that the flow stream is charged which means all type of cells will be charged. It makes sense to charge them according to the scattering spectrum observed, in that way we know which to sort or not. The charging is programmed according to scattered pulse values we want. Please do elaborate this otherwise I will pull all my remaining hair out.
Regards,
Akshit
In a cell sorter, particles are delivered in a buffered salt solution, capable conducting electrical current. We decide (based on the pattern of scatter and fluorescent signals) at the laser interrogation, if we want to sort a particle or not. We have no a bit of time to execute this sort decision, because the particle needs to travel from the laser interrogation to the droplet break off point. Once arrived there, we are charging the whole stream (here you´re right) and the leaving drop is carrying this charge and can be deflected. This works, because electrical current is travelling fast and is arriving at the same time at the break off point as the particle.
(PS: Watch the slide around minute 10 “Defining drop delay - Accu Drop”: In the bottom of the screenshot you see numbers with sliders for 2nd, 3rd and 4th drop. This is correcting for potential remaining charges on the stream, once a drop has left. It ensures, that the next drop has a net-charge of zero before it can be charged again for the next sort decision.)
first we decide if we want to sort this particle and in which bucket, then we apply positive or negative charge accordingly to the whole stream, then we wait for the droplet to break off, and then we ground the stream so it loses all charge and becomes neutral again until new sort
Great video!!! Thank you!
I wonder how this setup compares to Sphere Fluidics: ua-cam.com/video/AajNacI_HEo/v-deo.html
FACS sorting hehe
Cknc
Tfc nghkn😂😂😂😂