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Hi Lloyd,
I've found that the red and blue emission aren't quite lined up vertically,
at least with our "blue" 63x objective, so have to cut the top one or two
slices off a blue vertical series (depending on depth of slice), and the
bottom slices off a red series and reassemble to get the emissions aligned -
i.e. from the same depth in the tissue. The objectives do vary a bit,
perhaps ours isn't as well corrected as some.
cheers,
Rosemary
On 11/01/11 9:16 AM, "Lloyd Donaldson" <[log in to unmask]>
wrote:
> *****
> To join, leave or search the confocal microscopy listserv, go to:
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> *****
>
> Hi Rosemary
>
> We don't really work with live tissue but we do have a Leica 63x water lens on
> our old TCS NT. I never really found it particularly useful. I haven't tried a
> direct comparison on our new SP5 II but I would expect the glycerol lens to be
> much better unless you have to be in water.
> If your blue 63x is the same as ours I have imaged blue green and red both
> sequentially and simultaneously without any obvious problem. We image lignin
> at blue and or green emission and combine with alexa568 or alexa647 for immuno
> work. There is sometimes some quenching going on so we separate the lignin and
> alexa as far as we can. We have to mount in oil for this work which is why I
> am using the blue lens.
>
> Lloyd
>
> -----Original Message-----
> From: Confocal Microscopy List [mailto:[log in to unmask]] On
> Behalf Of Rosemary White
> Sent: Tuesday, 11 January 2011 10:48 a.m.
> To: [log in to unmask]
> Subject: Re: Glycerol Objectives - experience with
>
> *****
> To join, leave or search the confocal microscopy listserv, go to:
> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> *****
>
> Dear Lloyd,
>
> Does the 63x glycerol objective provide substantial improvements over the
> 63x water (with correction collar) for live tissues? We have one of the
> "blue" Leica 63x objectives, corrected for the blue-UV end of the spectrum
> (though only have 405 laser), which is great except when you want to image
> both red and blue emission....
>
> thanks,
> Rosemary White
>
>
> Dr Rosemary White
> CSIRO Plant Industry
> GPO Box 1600
> Canberra, ACT 2601
> Australia
>
> T 61 2 6246 5475
> F 61 2 6246 5334
> E [log in to unmask]
>
>
>
> On 11/01/11 6:21 AM, "Lloyd Donaldson" <[log in to unmask]>
> wrote:
>
>> *****
>> To join, leave or search the confocal microscopy listserv, go to:
>> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
>> *****
>>
>> Jeremy
>>
>> We have a Leica 63x glycerol objective with adjusting ring which gives very
>> noticeable improvement over 63x oil which we also have. We also have a 20x
>> multi immersion objective that can use glycerol. We have both oil and
>> glycerol
>> objectives, although we prefer glycerol as a mounting medium it is
>> incompatible with some of our samples because of induced swelling so we have
>> to mount in oil for those samples. We also have the oil lens because it is UV
>> compatible - we have a 355 nm laser.
>>
>>
>> Dr Lloyd Donaldson
>>
>> Senior Scientist, Project Leader - Microscopy/Wood Identification
>> Scion - Next Generation Biomaterials
>> Private Bag 3020, Rotorua
>> New Zealand 3010
>>
>> Ph: 64 7 343 5581
>>
>>
>>
>> -----Original Message-----
>> From: Confocal Microscopy List [mailto:[log in to unmask]] On
>> Behalf Of Jeremy Adler
>> Sent: Monday, 10 January 2011 9:39 p.m.
>> To: [log in to unmask]
>> Subject: Glycerol Objectives - experience with
>>
>> *****
>> To join, leave or search the confocal microscopy listserv, go to:
>> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
>> *****
>>
>> We are considering buying a new confocal for examining fixed and
>> potentially thick biological specimens.
>> Given the need to have to an RI match between the specimen, mounting
>> medium and immersion medium, I want to ask about the pros and cons of
>> glycerol objectives.
>>
>> Oil objectives appear to have no obvious advantages, the higher RI
>> will only apply if specimens are mounted in a medium with a RI close
>> to that of oil - while most media seem to have lower RIs. Why would
>> anyone choose an oil objective and which mounting media work for
>> thick specimens ?
>>
>> A possible disadvantage of glycerol objectives is that glycerol is
>> hygroscopic which could change its RI. Are there any RI equivalent
>> immersion media ?.
>>
>> So the pros and cons of oil or glycerol objectives.
>>
>>
>>
>>
>>
>>
>>
>> Jeremy Adler
>> Genetics & Pathology
>> Rudbeckslaboratoriet
>> Daghammersköljdsväg 20
>> 751 85 Uppsala
>> Sweden
>>
>> 0046 (0)18 471 4607
>>
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>
> Disclaimer: This e-mail and any attachments may contain information which is
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