Re: small and variable working distance

Posted by mmodel on
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That's right, that's how deep the water layer should be, and we do use all kinds of sandwiches, even without microspheres. But when you need to continuously exchange solutions, especially while keeping everything at 37C, it becomes a little inconvenient. A Bioptechs flow chamber with a thin spacer works well when cells can be grown on glass, but for cells on a plastic dish the simplest thing would be simply to bring the objective close enough to the bottom. The working distance of common water-immersion objectives is too large for that.

Mike
________________________________________
From: Confocal Microscopy List [[hidden email]] On Behalf Of Jeremy Adler [[hidden email]]
Sent: Friday, December 09, 2011 7:22 AM
To: [hidden email]
Subject: Re: small and variable working distance

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Out of interest, is the limitation that the water layer is only 20-30
microns thick or that the objective really needs to be that close ?

a 20-30 micron thick layer of water can easily by sandwiched between a
slide and a coverslip using microspheres as spaces. Then image with a
water objective that works thru a coverslip.




Quoting "MODEL, MICHAEL" <[hidden email]>:

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> 20-30 microns in water. I doubt such lenses exist but who knows... Thanks!
>
> Mike
>
> ________________________________________
> From: Confocal Microscopy List [[hidden email]] On
> Behalf Of Martin Wessendorf [[hidden email]]
> Sent: Thursday, December 08, 2011 9:20 PM
> To: [hidden email]
> Subject: Re: small and variable working distance
>
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> Dear Mike--
>
> How close do you need to get and what kind of resolution do you need?
> And in what medium?  Air?  Water?  Oil?
>
> Good luck!
>
> Martin
>
> On 12/8/2011 3:47 PM, MODEL, MICHAEL wrote:
>
>> Hi Martin - that's correct, I want to bring the objective very
>> close to the specimen, almost touching it.
>>
>> -----Original Message-----
>> From: Confocal Microscopy List
>> [mailto:[hidden email]] On Behalf Of Martin
>> Wessendorf
>> Sent: Thursday, December 08, 2011 4:27 PM
>> To: [hidden email]
>> Subject: Re: small and variable working distance
>> Dear Mike--
>>
>> On 12/8/2011 3:05 PM, MODEL, MICHAEL wrote:
>>
>>> Dear Confocal Microscopists - Does anyone of an objective with
>>> variable, down to zero, working distance? Thanks!
>>
>> I'm not sure I'm getting the concept--do you want an objective that
>> *needs* to be in contact with the specimen to be in focus?  --It might
>> help me if you explained a bit more about what you're trying to do.
>>
>> Thanks--
>>
>> Martin Wessendorf
>
> --
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