Re: [micro-manager-general] HCS plugin

[PhilippeP <pog...@un...>]

PEARSON Matthew wrote
> Could you describe a little more how you use DF for HCS.  Where do you
> travel to to "measure" on the plate and how many offsets do you use?  When
> you travel to each first position in a grid and replace it in the xy list
> do you "measure" DF at the same time?

Hi Matt,

First, sorry for the (very very) late answer. I did not see your post
earlier... 
Better late than never, right?... Hmmmm.

We happily use HCS with Zeiss DF. In brief:
1- Perform plate calibration;
2- Use 3-point-AF, by going to the 3 angles of the largest virtual triangle
that would encompass the different wells you want to acquire and focusing on
your cells. Add these 3 points in a list (via "+ Mark point" button), you
can then "Set 3-point list". 
You will see your chosen triangle appear on your virtual plate. You can now
delete the list, and create the real list with you grids. You will notice
that each position created has its own z value, computed from the plane
defined with your three point list. 
Even if focusing is not perfect on all points when doing "Go to" in the
list, you are usually well within the working range of the Definite Focus.
Perform your acquisition with DF selected in MDA, and your pictures should
be fine. Of course, you must first set your DF up!

Hope that helps,
Philippe

PS:
While I am at it, there is still no plan on having an option to create grids
in circular pattern so as not to "waste" cells that are not present in a
square grid? Sorry to just ask without proposing to do it. I wish I could.
But my coding abilities are close to null...





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