Search the CONFOCAL archive at
http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal
One possibility lies in combining holographic microscopy (see
http://www.lynceetec.com/downloads/ScientPubli/20051114_OX13_Rappaz.pdf)
with confocal microscopy.....although I don't know of anyone who has
tried it yet.
Bill Miller
At 12:15 PM 2/13/2007 +1100, Guy Cox wrote:
>Search the CONFOCAL archive at
>http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal
>
>It is rather an intractable problem. Without actual
>measurement useful correction is really pretty much
>impossible. People have tried schemes such as normalizing
>each slice but eventually this just brings up ever fainter
>out of focus images, which is not what you want.
>
>If there is some structure which you are confident is
>labelled uniformly you could use that as a standard and
>work out a correction, if not maybe you could put fluorescent
>beads on the top and bottom of your sample. You also need
>to allow for bleaching, so you should collect both a 'top-
>down' and a 'bottom-up' stack. (In principle, at least,
>multiphoton would avoid the need for that last step.)
>
> Guy
>
>Judy Trogadis wrote:
>>Search the CONFOCAL archive at
>>http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal
>>I am hoping to re-visit the issue of depth-related signal loss inherent
>>in confocal z-series and possible corrections. As I sit here and think
>>about it, I realize that it is an inevitable phenomenon, however, people
>>do use the intensity information collected from slices so there must be
>>some solution to this dilemma.
>>Some parameters we cannot control (e.g. the homogeneity of our tissue)
>>but of the ones we can, I would like to know what solutions people have
>>found. Is laser power a factor? Dye chemistry? Some confocals have
>>incorporated a correction factor into their software - is this fair? Is
>>correction ever possible after the data has been collected, as part of
>>the analysis (an Image J plugin)? Is signal loss less with a higher NA
>>objective?
>>Some colleagues are trying to do 3D modeling with quantitation of
>>microcapillaries and their algorithm is having major problems.
>>Thanks in advance
>>Judy
>>
>>Judy Trogadis
>>Bio-Imaging Coordinator
>>St. Michael's Hospital, 7Queen
>>30 Bond St.
>>Toronto, ON M5B 1W8, Canada
>>ph: 416-864-6060 x6337
>>pager: 416-685-9219
>>fax: 416-864-6043
>>[log in to unmask]
>
>--
>
>Optical Imaging Techniques in Cell Biology
>by Guy Cox CRC Press / Taylor & Francis
> http://www.guycox.com/optical.htm
>______________________________________________
>Associate Professor Guy Cox, MA, DPhil(Oxon)
>Electron Microscope Unit, Madsen Building F09,
>University of Sydney, NSW 2006
>______________________________________________
>Phone +61 2 9351 3176 Fax +61 2 9351 7682
>Mobile 0413 281 861
>______________________________________________
>http://www.guycox.net
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