Re: [micro-manager-general] HCS plugin

[Kurt T. <kur...@uc...>]

We have found that many plates are very far from flat - we have seen 
plates where the well bottom differs by 200 um over the surface of the 
plate.  This company makes plates that claim to be +/- 15 um over the 
plate: 
http://www.porvair-sciences.com/en/services-menu/life-sciences/glass-bottom-assay-plates/

We haven't tried them, but that might be a good starting point. Other 
companies may have similar products; I don't know.

Kurt

On 1/25/2016 8:26 AM, PEARSON Matthew wrote:
> Hi all,
>
> I was wondering how many of you use the High Content Screening plugin within Micro-Manager?  And whether you can auto-focus reliably? I have a plastic 96 well plate and struggling to achieve consistent focus using either the 3 point auto focus or JAF (H & P) methods.  There seems to be considerable Z offset between the wells and the multi-well plate insert and stage has no grub screws for adjusting any tilt.  I am imaging at 10x so perhaps at the limit of what the 3 point interpolation focus can deal with.  I have tested JAF (H&P) be defocussing the live image and trying different z steps and i have been able to get it to work sometimes like this but as soon as i set it to capture all the imaging sites, many are out of focus.  We may try using a glass bottomed plate which should at least decrease the error inherent in the plate itself.  I just wondered how much this is being used by the community for HCS.
>
> Thanks,
>
> Matt
>
>
> --
> Matt Pearson
> Microscopy Facility
> MRC Human Genetics Unit
> IGMM
> University of Edinburgh
> Crewe Road
> EH4 2XU
>
>
>
>
>


-- 
Kurt Thorn
Associate Professor
Director, Nikon Imaging Center
http://thornlab.ucsf.edu/
http://nic.ucsf.edu/blog/