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Glen
If you want to do a quick check to see if the stage and objective are
in parallel planes you can use this procedure.
1. Get two partially reflecting mirrors or beam splitters (percentage
age does not matter).
2. Place one laying flat on the hole in the nosepiece where the
objective belongs the other on the stage where the specimen belongs.
3. Remove the condenser, close down the field diaphragm all the way
and adjust the transmitted light source for comfort.
4. Remove the eyepieces and observe the alignment of the reflections
between the two mirrors.
If the reflections align you are good to go, if not, apply a little
pressure on the beamsplitter on the stage and observe the direction
of the reflection.
The relationship of the nosepiece and the stage mounts are supposed
to be set at the factory at time of assembly. However, due to
variations in all the supportive contact points between the specimen
plane and the shoulder of the objective threads it does not hurt to
check it once and a while.
Dan
On Mar 28, 2007, at 8:00 AM, Michelle Ocana wrote:
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Glen,
It sounds like the objective is contacting your slide. What is the
objective that you are using? I can tell you what the working
distance is.
Michelle
Michelle Ocana
Confocal and Imaging Specialist
Optical Analysis Corporation
3 Bud Way, Suite 25
Nashua, NH 03063-1700
c: 617-447-4391
-----Original Message-----
From: Confocal Microscopy List [mailto:[log in to unmask]]
On Behalf Of Glen MacDonald
Sent: Tuesday, March 27, 2007 3:45 PM
To: [log in to unmask]
Subject: Image shift
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Hi,
I'm trying to diagnose an odd problem with an FV-1000 and IX-81
microscope, to get speed things in getting this serviced. When
collecting a z-series, around 330 to 400 µm into the volume, the
image shifts in the x-axis by several pixels to the left, sometimes 1
or 2 lines upwards. The shift may occur within 1 line, or up to 15
lines resembling a vibration, but trending to the left. in the
subsequent portion of the stack, additional shifts will occur every
40 to 120 µm. If I capture the full volume in increments of less
than 400 µm, the shift will still occur. I've used box sizes from
256 to 1024 at dwell times of 2 µs to 20 µs. At 2 µs with a 256x256
box, the shift doesn't occur within a frame, but seems to be between
the frames. Also, the shift always occurs in the upper 1/3 of an image.
In line sequential acquisition, the shift appears in all channels.
With sequential frame acquisition, the shift appears within a frame
of the first channel but the slice images for the other channels
shift in their entirety.
The air table is up, although vibrations shouldn't be this regular.
Also, the starting point of the z-series may vary, relative to the
coverslip.
Any suggestions?
Regards,
Glen
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
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************************************************************************
******
The box said "Requires WindowsXP or better", so I bought a Macintosh.
************************************************************************
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Dan Focht
Bioptechs Inc.
V (724)282-7145
F (724)282-0745
www.bioptechs.com
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