Advice on deconvolution software for widefield microscope

[Pedro Camello]

Search the CONFOCAL archive at
http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal

Hi all

We´re considering purchase a deconvolution package to acquire images in a
TE2000 Nikon with a Orca-II ER camera. Deconvol would be oriented to resolve
subcellular structures at high magnification and tissue structures at low
(x10-x20) mag (if that´s possible). I need opinions regarding:

1. Best software
2. Best z control for the TE2000 (objectives or stage?)

I would also to know whether Z motorization is necessary only occasionally,
to calculate PSF, or is a used in each deconv-aimed acquisition.

Thanks in advance

  Pedro

[Sam's Mail]

Search the CONFOCAL archive at
http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal

Pedro,

If you would like to perform deconvolution well, then you will essentially need to utilize your Z-motor of choice for all of those image captures.  2D deconvolution is a technique that is most definitely not the same creature as robust 3D deconvolution methods, and your mileage will significantly vary with that approach.  When you refer to a stage motor, I assume you might be talking about a piezo-z stage from ASI or similar.  These are fantastic if you'd like to go extremely fast and don't have incredibly thick samples to work with.  If fast isn't an issue, than typically the precision from your built-in TE2000 motorization should be adequate for your work. As far as software choice goes, you'll find many different options that are quite viable.   I have had numerous successes at different times utilizing SlideBook from Intelligent Imaging Innovations (No commercial affiliation) for this purpose.  If I'm not mistaken company was initially created while solving some widefield deconvolution challenges, from a hardware and software angle. I have found that their team is knowledgeable about the issues surrounding this line of work, and may still be willing to do a retrofit as needed.  If you'd like more details, feel free to contact me offline.

--
Samuel A. Connell
Director of Light Microscopy
Cell & Tissue Imaging Center
St. Jude Children's Research Hospital
332 North Lauderdale St., E7061
Memphis, TN 38105-2794
Office [phone]
Cell [phone]
[hidden email]



________________________________________
From: Confocal Microscopy List [[hidden email]] On Behalf Of Pedro Camello [[hidden email]]
Sent: Monday, March 31, 2008 6:07 PM
To: [hidden email]
Subject: Advice on deconvolution software for widefield microscope.    .

Search the CONFOCAL archive at
http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal

Hi all

We´re considering purchase a deconvolution package to acquire images in a
TE2000 Nikon with a Orca-II ER camera. Deconvol would be oriented to resolve
subcellular structures at high magnification and tissue structures at low
(x10-x20) mag (if that´s possible). I need opinions regarding:

1. Best software
2. Best z control for the TE2000 (objectives or stage?)

I would also to know whether Z motorization is necessary only occasionally,
to calculate PSF, or is a used in each deconv-aimed acquisition.

Thanks in advance

  Pedro