Hi Yevgeniy,
I’ve never found using a level works either. I’ve assumed this is partly the lack of sensitivity and partly because we don’t care (in this case)
that the stage is level with the floor or “gravity” – only that it’s level with the *imaging plane*.
If I want / need the stage really flat, then I’ve used a method similar to Craig’s – i.e. imaging different positions on a known flat sample and
adjusting the insert each time through a few iterations.
This is probably easiest with something like this slide from Thorlabs (no commercial interest):
https://www.thorlabs.com/thorproduct.cfm?partnumber=R1L3S3P
Focus on one corner of the largest grid, then move the stage to another corner and adjust the stage insert levelling screws. Repeat this a few times
with each corner.
You could also use a multi-well plate (probably cheaper than the above if you have them lying around) for even more accuracy (i.e. larger distance
travelled by stage = more sensitive detection of focus shift). It’s easy enough to focus on the bottom of the wells by stopping down the condenser aperture to increase the contrast.
James
From: Confocal Microscopy List [mailto:[hidden email]]
On Behalf Of Craig Brideau
Sent: 28 October 2016 20:26
To: [hidden email]
Subject: Re: digital levels for microscope stages
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I have used the interference pattern off a coverslip to level a sample. You can also use a thin layer of a fluorescent material or a thin grid to make sure your stage is aligned with your imaging plane.
Craig
On Fri, Oct 28, 2016 at 12:56 PM, <[hidden email]> wrote:
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Hello everybody
Does anybody have any experience with using levels to make sure the stage inserts on their systems are as flat as possible? We use a little bubble level sometimes to adjust our stage inserts as well as make sure that the AFM head on one of our systems is flat. I feel like it's not sensitive enough. Would anybody be able to recommend a good digital level with high sensitivity that can be used for the purposes that I described.
Thanks very much in advance,
Yevgeniy Romin=====================================================================
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