Hanging Drop Aggregation Assay

[cardosopedro]

Hi.

I was reading some papers in which this method was employed but I don't understand one part of the procedure. Maybe this is because I'm missing something behind the basis of the method. Can someone explain me?

My doubt is: for quantification, some protocols say "Resulting cell clusters were subjected to 30 rounds of pipetting through a 200-ul Gilson pipette, and the degree of dissociation was quantified by counting the particles after trituration". I don't get this part...

So, after the incubation the drops/clusters of cells are still in the lid of the plate, right? What am I supposed to do with them?

And another question: how many drops per well should I put in the lid of a 24-well plate?

Thanks!