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Dear Jerry,
First, which version of LaserSharp and on what computer is it running?
If you are are still on the original Compaq prosignia, I strongly
recommend contacting Bio-Rad about replacement with the Dell PowerEdge,
and running LS 3.2, for far fewer crashes.

The minimum box height with LS 3.2 seems to be 8 lines.  That gives
about 16 scans/sec using for an XYT scan mode with the interval set to
'0' and holding the shutter open between scans.  Box width has no
effect on scan time.  a 512x8 box gives the same scan rate, and also
this is independent of zoom.    I tried making a method but crashed,
learning that there exists a limit of 30 methods.  A rather crude
notice.

However, if you have LS 3.2 with Timecourse, then simply set up a new
timecourse and select the XT option, leaving your box size at 512x512.
Set the Y position of the line and choose the 'save every image'
option.  click the button for timing and background, then click on
Record. you will get a stack of sequential linescan images.

Hope this helps.
Regards,

Glen


On Tuesday, November 19, 2002, at 07:08 PM, Jerry Sedgewick wrote:

> Search the CONFOCAL archive at
> http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal
>
> I am attempting to image at 2 ms resolution on a Bio-Rad 1024, only
> because
> I read that it could be done in the accompanying Bio-Rad manuals.  Fast
> imaging was also alluded to by the person who sold this device to our
> core
> facility years ago (I was told a line scan should give me 100 frames
> per
> second).
>
> At any rate, I have created a method for single wavelength (not ratio)
> imaging using Fluo-3, employing only one PMT.  In the time course
> component
> of the Laser Sharp software, I have chosen XT line scanning with a box
> size
> at its smallest (160X160?).  The user-defined box I had hoped to make
> resulted only in crashing the computer.  I zoomed in to create what was
> listed as an 80 micron line.  I did not collect images at all, and the
> scan
> rate was set at Normal.  For all practical purposes, these settings
> should
> have resulted in something closer to 2 ms resolution than the best I
> was
> able to get: ~3 scans per second.
>
> I had hoped to get an answer from technical help at Bio-Rad, but they
> simply gave me wrong advice.  So I appeal to this group to determine
> whether I should expect scan rates necessary for calcium spark
> acquisition
> with a 1024 (and, if so, what is the elusive thing I have
> overlooked?), or
> whether I should bag it.
>
> Jerry
>
> Jerry Sedgewick
> Program Director
> BIPL (Biomedical Image Processing Lab)
> 312 Church St. SE, 1-205 BSBE
> Minneapolis, MN  55455
> [log in to unmask]
> www.bipl.ahc.umn.edu
> www.quickphotoshop.com
> 612-624-6607
>
Glen MacDonald
Core for Communication Research
Virginia Merrill Bloedel Hearing Research Center
Box 357923
University of Washington
Seattle, WA 98195-7923  USA
(206) 616-4156
[log in to unmask]

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The box said "Requires Windows 95 or better", so I bought a Macintosh.
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