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> 1- I am looking for the publication or book (or any reference) which shows
> that
> a correctly acquired image has a histogram distribution of pixel intensity
> with
> a count of 0 at intensities 0 and 255.  I know I read this somewhere...

I'm sure that there are many sources for this.  You will
find it in Chapter 6 of my book 'Optical Imaging Techniques
in Cell Biology' (CRC Press / Taylor & Francis) and it must
also be in Jim Pawley's encyclopaedic Handbook of Biological
Confocal Microscopy.

> 2- I have a technical question about ensuring that the detector is not
> saturated
> during acquisition.  I usually set the laser power and detector gain with
> a 1
> second scan (512x512 pixels) and then select averaging to collect the
> final
> image.  If I were to select a longer dwell time during set up, I may
> select a
> different gain setting for the detector because saturated pixels are less
> probable (but did I saturate the detector during the scan...). So how is
> the
> signal being processed for each pixel?

If you select averaging the final result should be
essentially equal to to single scan value since averaging
is summing the signal then dividing by the number of frames.
So you should certainly NOT change dwell times after setting
contrast and black level.



> 3- I have a further technical question about the Zeiss LSM410.  Are line
> and
> frame averaging both using the Kalman averaging approach?

I don't know too much about the Zeiss but in the Leica they
are equivalent and it's hard to see how they could not be.

                                                 Guy


-- 
Associate Professor Guy Cox
Electron Microscope Unit,
University of Sydney,
NSW 2006, Australia

Phone:+61 2 9351 3176    Fax:+61 2 9351 7682
http://www.guycox.net