Invitation to Vendor Webinar - Advances in Super-resolution Microscopy

[Clements, Ian]

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Greetings and a Happy New Year to all list members.

I would like to extend the following invitation to join our Free webinar on "Advances in Super-Resolution Microscopy" on January 19, 2011.  The webinar will be hosted at 3 times, just register for the event that is most convenient.

6 a.m. PST
11 a.m. PST
6 p.m. PST

Register at http://apiwebinars.webex.com

During this informative webinar, we will cover:
* Methods for overcoming diffraction limited microscopy
* New super-resolution capabilities on the DeltaVision OMX platform
* Making live cell super-resolution imaging a reality

For more information, please e-mail us at [hidden email].

Ian Clements
Product Manager - DeltaVision OMX Systems


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This email message, together with any attachments, is for the sole use of the intended recipient(s) and is the confidential information of Applied Precision Inc. If you are not the intended recipient, your review, use, disclosure, copying or dissemination of this email message or its attachments, or the information contained therein, is strictly prohibited. If you are not the intended recipient or if you think this email was sent to you in error, please notify the sender by reply email and delete this message and its attachments, as well as all copies, from your system.

[Jeremy Adler-3]

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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

[Adrian Smith-6]

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On related note our Olympus rep just pointed me to their new silicone oil objectives...

http://www.microscopy-analysis.com/news/olympus-introduces-uplsapo-30x-and-60x-silicone-oil-immersion-objectives

http://microscope.olympus-global.com/en/ga/product/fv1000/sf03.cfm
http://microscope.olympus-global.com/uis2/en/uplsapo30xs/
http://microscope.olympus-global.com/uis2/en/uplsapo60xs/

Anyone tried one?

Regards,

Adrian

On 10/01/2011, at 7:39 PM, Jeremy Adler wrote:

> *****
> 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

______________________________________________
Dr Adrian Smith
    Manager, Cytometry & Imaging Facilities      
    Centenary Institute
    http://www.centenary.org.au     
    Locked Bag No.6 Newtown, NSW 2042 AUSTRALIA.                                                                    
    Ph: 61-2-9565-6189     Fax: [phone]




______________________________________________
 Dr Adrian Smith
     Manager, Cytometry & Imaging Facilities      
     Centenary Institute
     http://www.centenary.org.au     
     Locked Bag No.6 Newtown, NSW 2042 AUSTRALIA.                                                                    
     Ph: 61-2-9565-6189     Fax: [phone]

[Craig Brideau]

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We have a Nikon 20x multi-medium lens that works quite well.  It works with
air/water/glycerol or oil.  We've even used it for decent 2-photon pictures.
 It has a collar for adjusting to different mediums, and fine adjustment of
the collar can improve the image a bit by correcting for some aberration.
 One side bonus is that since it is rated for oil or water, it doesn't
matter if it accidentally gets either on it.  We primarily use it in either
water or oil regimes.  We have used it as a water immersion lens to image
samples without coverslips.  For samples with coverslips we use oil.  Oil
does allow for a higher NA, if your sample is right on the surface of the
coverslip.  Again, fine adjustment of the correction collar helps here.

Craig


On Mon, Jan 10, 2011 at 1:39 AM, Jeremy Adler <[hidden email]>wrote:

> *****
> 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
>

[Lloyd Donaldson]

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To join, leave or search the confocal microscopy listserv, go to:
http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
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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:[hidden email]] On Behalf Of Jeremy Adler
Sent: Monday, 10 January 2011 9:39 p.m.
To: [hidden email]
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

Disclaimer: This e-mail and any attachments may contain information which is confidential or subject to copyright. If you receive this e-mail in error, please delete it.
Scion does not accept responsibility for anything in this e-mail which is not provided in the  course of Scion's usual business or for any computer virus, data corruption, interference or delay arising from this e-mail.

[Ignatius, Mike-2]

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I was told at SFN that one of the reasons for silicon immersion oil, is that glycerol changes its RI during prolonged imaging sessions.  Anyone know who much or how soon?

If it is significant, it is a good argument for silicon corrected lens - especially if doing long 3D reconstructions of thick tissue.

Mike Ignatius,
Molecular Probes/LifeTechnology

No commercial interest.


-----Original Message-----
From: Confocal Microscopy List [mailto:[hidden email]] On Behalf Of Lloyd Donaldson
Sent: Monday, January 10, 2011 11:22 AM
To: [hidden email]
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
*****

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:[hidden email]] On Behalf Of Jeremy Adler
Sent: Monday, 10 January 2011 9:39 p.m.
To: [hidden email]
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

Disclaimer: This e-mail and any attachments may contain information which is confidential or subject to copyright. If you receive this e-mail in error, please delete it.
Scion does not accept responsibility for anything in this e-mail which is not provided in the  course of Scion's usual business or for any computer virus, data corruption, interference or delay arising from this e-mail.

[Rosemary.White]

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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 [hidden email]



On 11/01/11 6:21 AM, "Lloyd Donaldson" <[hidden email]>
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:[hidden email]] On
> Behalf Of Jeremy Adler
> Sent: Monday, 10 January 2011 9:39 p.m.
> To: [hidden email]
> 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
>
> Disclaimer: This e-mail and any attachments may contain information which is
> confidential or subject to copyright. If you receive this e-mail in error,
> please delete it.
> Scion does not accept responsibility for anything in this e-mail which is not
> provided in the  course of Scion's usual business or for any computer virus,
> data corruption, interference or delay arising from this e-mail.

[Lloyd Donaldson]

*****
To join, leave or search the confocal microscopy listserv, go to:
http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
*****

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:[hidden email]] On Behalf Of Rosemary White
Sent: Tuesday, 11 January 2011 10:48 a.m.
To: [hidden email]
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 [hidden email]



On 11/01/11 6:21 AM, "Lloyd Donaldson" <[hidden email]>
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:[hidden email]] On
> Behalf Of Jeremy Adler
> Sent: Monday, 10 January 2011 9:39 p.m.
> To: [hidden email]
> 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
>
> Disclaimer: This e-mail and any attachments may contain information which is
> confidential or subject to copyright. If you receive this e-mail in error,
> please delete it.
> Scion does not accept responsibility for anything in this e-mail which is not
> provided in the  course of Scion's usual business or for any computer virus,
> data corruption, interference or delay arising from this e-mail.

Disclaimer: This e-mail and any attachments may contain information which is confidential or subject to copyright. If you receive this e-mail in error, please delete it.
Scion does not accept responsibility for anything in this e-mail which is not provided in the  course of Scion's usual business or for any computer virus, data corruption, interference or delay arising from this e-mail.

[Rosemary.White]

*****
To join, leave or search the confocal microscopy listserv, go to:
http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
*****

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" <[hidden email]>
wrote:

> *****
> To join, leave or search the confocal microscopy listserv, go to:
> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> *****
>
> 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:[hidden email]] On
> Behalf Of Rosemary White
> Sent: Tuesday, 11 January 2011 10:48 a.m.
> To: [hidden email]
> 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 [hidden email]
>
>
>
> On 11/01/11 6:21 AM, "Lloyd Donaldson" <[hidden email]>
> 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:[hidden email]] On
>> Behalf Of Jeremy Adler
>> Sent: Monday, 10 January 2011 9:39 p.m.
>> To: [hidden email]
>> 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
>>
>> Disclaimer: This e-mail and any attachments may contain information which is
>> confidential or subject to copyright. If you receive this e-mail in error,
>> please delete it.
>> Scion does not accept responsibility for anything in this e-mail which is not
>> provided in the  course of Scion's usual business or for any computer virus,
>> data corruption, interference or delay arising from this e-mail.
>
> Disclaimer: This e-mail and any attachments may contain information which is
> confidential or subject to copyright. If you receive this e-mail in error,
> please delete it.
> Scion does not accept responsibility for anything in this e-mail which is not
> provided in the  course of Scion's usual business or for any computer virus,
> data corruption, interference or delay arising from this e-mail.

[Craig Brideau]

*****
To join, leave or search the confocal microscopy listserv, go to:
http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
*****

That's odd.  Doesn't Zeiss have a blue correction optic in the microscope to
keep things chromatically aligned?

Craig


On Mon, Jan 10, 2011 at 3:32 PM, Rosemary White <[hidden email]>wrote:

> *****
> To join, leave or search the confocal microscopy listserv, go to:
> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> *****
>
> 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" <[hidden email]>
> wrote:
>
> > *****
> > To join, leave or search the confocal microscopy listserv, go to:
> > http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> > *****
> >
> > 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:[hidden email]]
> On
> > Behalf Of Rosemary White
> > Sent: Tuesday, 11 January 2011 10:48 a.m.
> > To: [hidden email]
> > 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 [hidden email]
> >
> >
> >
> > On 11/01/11 6:21 AM, "Lloyd Donaldson" <
> [hidden email]>
> > 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:[hidden email]]
> On
> >> Behalf Of Jeremy Adler
> >> Sent: Monday, 10 January 2011 9:39 p.m.
> >> To: [hidden email]
> >> 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
> >>
> >> Disclaimer: This e-mail and any attachments may contain information
> which is
> >> confidential or subject to copyright. If you receive this e-mail in
> error,
> >> please delete it.
> >> Scion does not accept responsibility for anything in this e-mail which
> is not
> >> provided in the  course of Scion's usual business or for any computer
> virus,
> >> data corruption, interference or delay arising from this e-mail.
> >
> > Disclaimer: This e-mail and any attachments may contain information which
> is
> > confidential or subject to copyright. If you receive this e-mail in
> error,
> > please delete it.
> > Scion does not accept responsibility for anything in this e-mail which is
> not
> > provided in the  course of Scion's usual business or for any computer
> virus,
> > data corruption, interference or delay arising from this e-mail.
>

[mcammer]

*****
To join, leave or search the confocal microscopy listserv, go to:
http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
*****

A few years ago we evaluated the Leica 63X glycerol objective side-by-side with the water immersion one on the AOBS SP2 for work with live tissue and found the glycerol one to be enough of an improvement to purchase it.
-Michael C.

-----Original Message-----
From: Confocal Microscopy List [mailto:[hidden email]] On Behalf Of Rosemary White
Sent: Monday, January 10, 2011 4:48 PM
To: [hidden email]
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 [hidden email]



On 11/01/11 6:21 AM, "Lloyd Donaldson" <[hidden email]>
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:[hidden email]] On
> Behalf Of Jeremy Adler
> Sent: Monday, 10 January 2011 9:39 p.m.
> To: [hidden email]
> 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
>
> Disclaimer: This e-mail and any attachments may contain information which is
> confidential or subject to copyright. If you receive this e-mail in error,
> please delete it.
> Scion does not accept responsibility for anything in this e-mail which is not
> provided in the  course of Scion's usual business or for any computer virus,
> data corruption, interference or delay arising from this e-mail.

------------------------------------------------------------
This email message, including any attachments, is for the sole use of the intended recipient(s) and may contain information that is proprietary, confidential, and exempt from disclosure under applicable law. Any unauthorized review, use, disclosure, or distribution is prohibited. If you have received this email in error please notify the sender by return email and delete the original message. Please note, the recipient should check this email and any attachments for the presence of viruses. The organization accepts no liability for any damage caused by any virus transmitted by this email.
=================================

[Lloyd Donaldson]

*****
To join, leave or search the confocal microscopy listserv, go to:
http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
*****

Hi Rosemary

Yes I agree with you. There is sometimes some offset in z but I make a projection and then shift the red channel in xy if there is any obvious mis-alignment. Only have to do this occasionally though.

Craig
Yes Leica have a correction lens but it is for UV not blue. I would say chromatic alignment is much better for a UV/red combination but haven't tried UV/far red.

-----Original Message-----
From: Confocal Microscopy List [mailto:[hidden email]] On Behalf Of Craig Brideau
Sent: Tuesday, 11 January 2011 11:38 a.m.
To: [hidden email]
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
*****

That's odd.  Doesn't Zeiss have a blue correction optic in the microscope to
keep things chromatically aligned?

Craig


On Mon, Jan 10, 2011 at 3:32 PM, Rosemary White <[hidden email]>wrote:

> *****
> To join, leave or search the confocal microscopy listserv, go to:
> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> *****
>
> 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" <[hidden email]>
> wrote:
>
> > *****
> > To join, leave or search the confocal microscopy listserv, go to:
> > http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> > *****
> >
> > 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:[hidden email]]
> On
> > Behalf Of Rosemary White
> > Sent: Tuesday, 11 January 2011 10:48 a.m.
> > To: [hidden email]
> > 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 [hidden email]
> >
> >
> >
> > On 11/01/11 6:21 AM, "Lloyd Donaldson" <
> [hidden email]>
> > 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:[hidden email]]
> On
> >> Behalf Of Jeremy Adler
> >> Sent: Monday, 10 January 2011 9:39 p.m.
> >> To: [hidden email]
> >> 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
> >>
> >> Disclaimer: This e-mail and any attachments may contain information
> which is
> >> confidential or subject to copyright. If you receive this e-mail in
> error,
> >> please delete it.
> >> Scion does not accept responsibility for anything in this e-mail which
> is not
> >> provided in the  course of Scion's usual business or for any computer
> virus,
> >> data corruption, interference or delay arising from this e-mail.
> >
> > Disclaimer: This e-mail and any attachments may contain information which
> is
> > confidential or subject to copyright. If you receive this e-mail in
> error,
> > please delete it.
> > Scion does not accept responsibility for anything in this e-mail which is
> not
> > provided in the  course of Scion's usual business or for any computer
> virus,
> > data corruption, interference or delay arising from this e-mail.
>

Disclaimer: This e-mail and any attachments may contain information which is confidential or subject to copyright. If you receive this e-mail in error, please delete it.
Scion does not accept responsibility for anything in this e-mail which is not provided in the  course of Scion's usual business or for any computer virus, data corruption, interference or delay arising from this e-mail.

[Martin Wessendorf-2]

*****
To join, leave or search the confocal microscopy listserv, go to:
http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
*****

On 1/10/2011 4:32 PM, Rosemary White wrote:

> 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.

Can anyone explain where the difficulty arises in correcting for axial
chromatic aberration?  I've consistently seen serious axial chromatic
aberration in good quality oil objectives from one very reputable
manufacturer, between red, green and far-red (1 um off in the z-axis
between green and red, and between red and far-red; 2 um between green
and far-red).  I have always heard that axial chromatic aberration was
easy to correct for and would've thought that a solution for 3-color
correction would've been found 100 years ago.  However, I don't know
enough optics to understand the subtleties.  Are there trade-offs to
trying to obtain good correction, besides the cost in scattering of
adding additional lens elements?

Thanks--

Martin
--
Martin Wessendorf, Ph.D.                   office: [phone]
Assoc Prof, Dept Neuroscience                 lab: [phone]
University of Minnesota             Preferred FAX: [phone]
6-145 Jackson Hall, 321 Church St. SE    Dept Fax: [phone]
Minneapolis, MN  55455                    e-mail: [hidden email]

[Craig Brideau]

*****
To join, leave or search the confocal microscopy listserv, go to:
http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
*****

To design a chromatically corrected optical system you need to use multiple
optics made of different types of glass.  The gist of it is that each glass
has slightly different wavelength-dependent indecies of refraction.  When
you put them together in the right way you can partly cancel out the
chromatically sensitive effects of the system.  The problem is that you can
usually only nail down two or three specific discrete wavelengths at a time.
 The simplest lens for this is an achromatic doublet.  It uses two types of
glass and has the same focal point for two specific wavelengths.  Operating
away from these two wavelengths will cause the focal point to drift.  The
better the design the less drift occurs, but it is really only 'perfect' for
two very specific wavelengths.  A triplet uses three pieces of glass
(typically) and nails down three specific wavelengths.  Basically you are
limited to designing for specific wavelengths.  The difficulty becomes which
specific wavelengths do you want to design for?  You can get 'good'
correction over a range, but you can only be dead-on for discrete values.
 You can get 'close' over a fairly wide range, but it takes a lot of design
work to pull this off.  This is further complicated by the fact that most
glasses have a steep change in optical properties towards the blue/violet/UV
portion of the spectrum, so if you are working in that range it becomes
trickier.  The balance between the different glass types and their
interactions with the overall design are highly complex, so as you make the
lens better it becomes astronomically more complicated and thus expensive.

Craig


On Mon, Jan 10, 2011 at 4:18 PM, Martin Wessendorf <[hidden email]> wrote:

> *****
> To join, leave or search the confocal microscopy listserv, go to:
> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> *****
>
> On 1/10/2011 4:32 PM, Rosemary White wrote:
>
>  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.
>>
>
> Can anyone explain where the difficulty arises in correcting for axial
> chromatic aberration?  I've consistently seen serious axial chromatic
> aberration in good quality oil objectives from one very reputable
> manufacturer, between red, green and far-red (1 um off in the z-axis between
> green and red, and between red and far-red; 2 um between green and far-red).
>  I have always heard that axial chromatic aberration was easy to correct for
> and would've thought that a solution for 3-color correction would've been
> found 100 years ago.  However, I don't know enough optics to understand the
> subtleties.  Are there trade-offs to trying to obtain good correction,
> besides the cost in scattering of adding additional lens elements?
>
> Thanks--
>
> Martin
> --
> Martin Wessendorf, Ph.D.                   office: [phone]
> Assoc Prof, Dept Neuroscience                 lab: [phone]
> University of Minnesota             Preferred FAX: [phone]
> 6-145 Jackson Hall, 321 Church St. SE    Dept Fax: [phone]
> Minneapolis, MN  55455                    e-mail: [hidden email]
>

[George McNamara]

*****
To join, leave or search the confocal microscopy listserv, go to:
http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
*****

Hi Jeremy,

A lot cheaper to optimize the specimen refractive index tl an oil
immersion lens.

$63.20 ... 2,2'-thiodiethanol, 1 liter (Staudt ... Hell, Microsc Res Tech).

Pricing from one of the microscope companies (other companies will be
similar):

$12,445 ... 63x/1.2 water w/correction collar (coverglass thickness 0.14
to 0.19 mm), apochromat

$10,732 ... 63x/1.3 Imm Corr DIC (corr = coverglass thickness 0.15 to
0.19 mm)

$6,723 or $7,065 ... 63x/1.4 oil iris NA 0.7 to 1.4), plan apochromat

$6,075 ... 63x/1.4 oil DIC (for coverglass 0.17 mm)

So, you can buy the last objective lens (DIC prisms not included) and a
whole lot of TDE for a lot less than the other lenses. See Stan Vitha's
email message to the listserv for his recommendation on gradual
transition into TDE. Ignoring bulk volume discount, you could buy 1,000
bottles of 1 liter TDE plus the last objective lens for about the same
price as the first lens listed above. It will be a long time to use up
1,000 liters of TDE!

Philippe Clemenceau <[hidden email]> sent me a PDF of his
company's adaptive optics device for microscopy (he sent an email to the
listserv recently). Ask him for it. See also
http://www.imagine-optic.com/iop_applications_adaptive-optics_microscopy-life-sciences_en.php
I am looking forward to finding out the pricing (and compatibility with
cameras and imaging software) for use with improving standard widefield
fluorescence images. Could be a lot more useful than the manual (and
therefore pretty useless for thick specimens) correction collars on
various lenses. Philippe mentioned that his company's device can enable
some types of 3D PALM/STORM, and also reduce spherical aberrations for
both widefield fluorescence microscopy and biplane FPALM nanoscopy. 3i
has also been offering the "SAC - spherical aberration corrector" for a
couple of years.

enjoy,

George
TDE price from
http://www.sigmaaldrich.com/catalog/Lookup.do?N5=All&N3=mode+matchpartialmax&N4=thiodiethanol&D7=0&D10=thiodiethanol&N1=S_ID&ST=RS&N25=0&F=PR 



On 1/10/2011 3:39 AM, Jeremy Adler wrote:

> *****
> 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
>

[Andreas Bruckbauer]

*****
To join, leave or search the confocal microscopy listserv, go to:
http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
*****


 Hi,
a year ago Olympus has launched a  60x lens which is corrected for 405 nm in contrast to the 435 nm commenly corrected for.
http://www.microscopy-analysis.com/news/olympus-launches-super-corrected-60x-14-na-objective-lensc
They claim 0.1 - 0.2 micrometer chromatic shift from 405 - 650 nm.


 best wishes

Andreas


 

 

-----Original Message-----
From: George McNamara <[hidden email]>
To: [hidden email]
Sent: Tue, 11 Jan 2011 2:17
Subject: Re: Glycerol Objectives - experience with


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Hi Jeremy,
 
A lot cheaper to optimize the specimen refractive index tl an oil immersion lens.
 
$63.20 ... 2,2'-thiodiethanol, 1 liter (Staudt ... Hell, Microsc Res Tech).
 
Pricing from one of the microscope companies (other companies will be similar):
 
$12,445 ... 63x/1.2 water w/correction collar (coverglass thickness 0.14 to 0.19 mm), apochromat
 
$10,732 ... 63x/1.3 Imm Corr DIC (corr = coverglass thickness 0.15 to 0.19 mm)
 
$6,723 or $7,065 ... 63x/1.4 oil iris NA 0.7 to 1.4), plan apochromat
 
$6,075 ... 63x/1.4 oil DIC (for coverglass 0.17 mm)
 
So, you can buy the last objective lens (DIC prisms not included) and a whole lot of TDE for a lot less than the other lenses. See Stan Vitha's email message to the listserv for his recommendation on gradual transition into TDE. Ignoring bulk volume discount, you could buy 1,000 bottles of 1 liter TDE plus the last objective lens for about the same price as the first lens listed above. It will be a long time to use up 1,000 liters of TDE!
 
Philippe Clemenceau <[hidden email]> sent me a PDF of his company's adaptive optics device for microscopy (he sent an email to the listserv recently). Ask him for it. See also http://www.imagine-optic.com/iop_applications_adaptive-optics_microscopy-life-sciences_en.php 
I am looking forward to finding out the pricing (and compatibility with cameras and imaging software) for use with improving standard widefield fluorescence images. Could be a lot more useful than the manual (and therefore pretty useless for thick specimens) correction collars on various lenses. Philippe mentioned that his company's device can enable some types of 3D PALM/STORM, and also reduce spherical aberrations for both widefield fluorescence microscopy and biplane FPALM nanoscopy. 3i has also been offering the "SAC - spherical aberration corrector" for a couple of years.
 
enjoy,
 
George
TDE price from http://www.sigmaaldrich.com/catalog/Lookup.do?N5=All&N3=mode+matchpartialmax&N4=thiodiethanol&D7=0&D10=thiodiethanol&N1=S_ID&ST=RS&N25=0&F=PR 
 
On 1/10/2011 3:39 AM, Jeremy Adler wrote:

> *****
> 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
>

 

[Jerry (Gerald) Sedgewick]

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Hello All,

To all users of scientific imaging systems: If you or users of your
facility could benefit from a free forum on scientific imaging and image
analysis, I'm hosting one.  This is not a listserv like the Confocal
Microscopy group, but a web forum to give users the option of uploading
images for evaluation.  For more information and to register, go to:

http://www.imagingandanalysis.com/instruction.html

Cheers,

Jerry

--

Jerry Sedgewick
Sedgewick Initiatives
965 Cromwell Avenue
Saint Paul, MN  55114
651-788-2261
[hidden email]
http://www.imagingandanalysis.com

Author of: "Scientific Imaging with Photoshop: Methods, Measurement, and
Output"

[Ian Dobbie]

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Adrian Smith <[hidden email]> writes:

Sorry I'm a bit (3 months!) late to this discussion.

> On related note our Olympus rep just pointed me to their new silicone oil objectives...
>
> http://www.microscopy-analysis.com/news/olympus-introduces-uplsapo-30x-and-60x-silicone-oil-immersion-objectives
>
> Anyone tried one?

Yes we have the 60x SI objective and use it extensively to look at
Drosophila oocytes and embryos in halocarbon oil. It is especial
effective on our spinning disk system, where is leads to massive signal
increase compared to either an oil or water immersion objective.

Ian
--
Ian Dobbie
Micron Imaging Facility Manager,
Biochemistry,
University of Oxford,
South Parks Road,
Oxford
OX1 3QU
Tel: [phone]
Email: [hidden email]