Simple animal demo specimen ideas

[Stephen Cody-2]

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Dear Esteban,

I'm Not really sure what you are asking. I've not done rigorous tests with different NAs and RI mismatching. I've done a few comparisons for illustrative purposes.

But chances are if you are buying an expensive dipping lens you'll want to go as deep into tissue as possible. Confocal is very sensitive to aberrations. Spherical and longitudinal chromatic aberrations will cause the signal to be rejected by the confocal pinhole. This will get worse with depth.

If you are going to spend lots of $$$ on a lens, why compromise performance by using a lens from a different manufacturer which may do the chromatic aberration correction differently. You may end up with no correction or too much.

With any optical zoom system it is the NA that is important. This will limit the resolution. Two similar lenses with same NA . Eg 20 x, zoom 2 NA=1 compared with 40 x, zoom 1 NA=1
Should have identical performance, provided everything else is the same.

So why would you buy the 40x when the 20 x will do the job of both these objectives?

Of course you need to compare many other things, such as WD, flatness, chromatic aberration correction etc etc.

I hope that makes more sense.

Steve

Stephen H. Cody

On 05/03/2011, at 10:45 AM, "G. Esteban Fernandez" <[hidden email]> wrote:

> *****
> To join, leave or search the confocal microscopy listserv, go to:
> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> *****
>
> Steve,
>
> Just to clarify, you think the loss of resolution/signal is more with
> chromatic aberration than by reducing the NA by ~0.2 plus air/water RI
> mismatch?
>
> Thanks,
> Esteban
>
> On Fri, Mar 4, 2011 at 2:46 PM, Stephen Cody <[hidden email]> wrote:
>
>> *****
>> To join, leave or search the confocal microscopy listserv, go to:
>> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
>> *****
>>
>> G'day Aleksandr,
>>
>> Do you require this lens for confocal and or multiphoton microscopy? If so
>> you should follow George's advice.
>>
>> If you purchase a high NA 20x (or 25x) lens. Then this will perform
>> perfectly at zoom 1 with a wide field of view, and as described be George
>> zoom in 2 or 3 times and have the equivalent performance of a 40 or 60x.
>> Saving you the cost of several lenses.
>>
>> If however if you need a 40x for conventional microscopy, then you may need
>> a 40x.
>>
>> Mismatching objectives from manufactures may be possible, but is not a good
>> idea, especially if you want to use confocal microscopy to optically section
>> deep within tissue. The loss of chromatic correction I think would result in
>> very significant loss of signal strength as you go deeper into tissue with
>> confocal. Maybe not so problematic if using for conventional microscopy or
>> multiphoton with NDDs.
>>
>> I wouldn't mix manufactures lenses on confocal if you were hoping for good
>> depth penetration. Which is likely the reason why you want a water immersion
>> lens in the first place.
>>
>> Cheers Steve
>>
>> Stephen H. Cody
>>
>> On 26/02/2011, at 11:43 AM, George McNamara <[hidden email]>
>> wrote:
>>
>>> *****
>>> To join, leave or search the confocal microscopy listserv, go to:
>>> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
>>> *****
>>>
>>> Leica has a 20x/1.0 NA objective lens, and, if I recall correctly, a
>> 25x/0.95NA - if this is on a laser scanning confocal, zoom these lenses to
>> 2x. NA is what matters.
>>>
>>> On 2/25/2011 10:10 AM, Jurkevic, Aleksandr wrote:
>>>> *****
>>>> To join, leave or search the confocal microscopy listserv, go to:
>>>> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
>>>> *****
>>>>
>>>> Hello,
>>>>
>>>> Leica doesn't currently have a 40x water immersion objective although I
>> heard that they have one in the pipeline. Has anyone used a 40x water from a
>> different manufacturer with success on this frame?
>>>>
>>>>
>>>> Aleksandr Jurkevic, PhD
>>>> Associate Director
>>>> Molecular Cytology Core
>>>> University of Missouri-Columbia
>>>> 120 Life Science Center
>>>> 1201 E. Rollins St.
>>>> Columbia, MO 65211-7310
>>>>
>>>>
>>>
>>>
>>> --
>>>
>>>
>>> George McNamara, PhD
>>> Analytical Imaging Core Facility
>>> University of Miami
>>

[Guy Cox-2]

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This thread has got very convoluted.  Leica have their 'beer-can' 20x
water immersion which I think is NA 1.0 - we don't actually have that
lens so can't comment directly on it, but it's a mighty impressive chunk
of optical glass.  They also have a x63 water immersion, NA 1.2, which
we do have.  Surely one or other of these will meet any likely
requirement?  To get Nyquist resolution from an NA 1.2 lens you could
not be at zoom 1 at 40x anyway.  So if you really need the low
magnification you will be just as well off at NA 1.0.   Otherwise, x63
isn't exactly a huge gap from x40.   The rationale for using a
mis-corrected lens from another maker just isn't there.  

 

 
Guy

 

Optical Imaging Techniques in Cell Biology

by Guy Cox    CRC Press / Taylor & Francis

     http://www.guycox.com/optical.htm
<http://www.guycox.com/optical.htm>

______________________________________________

Associate Professor Guy Cox, MA, DPhil(Oxon)

Australian Centre for Microscopy & Microanalysis,

Madsen Building F09, University of Sydney, NSW 2006

 

Phone [phone]     Fax [phone]

             Mobile [phone]

______________________________________________

      http://www.guycox.net <http://www.guycox.net>

 

 

From: Confocal Microscopy List [mailto:[hidden email]]
On Behalf Of Stephen Cody
Sent: Saturday, 5 March 2011 7:59 PM
To: [hidden email]
Subject: Re: 40x water immersion objective on Leica miscroscope

 

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

Dear Esteban,

I'm Not really sure what you are asking. I've not done rigorous tests
with different NAs and RI mismatching. I've done a few comparisons for
illustrative purposes.

But chances are if you are buying an expensive dipping lens you'll want
to go as deep into tissue as possible. Confocal is very sensitive to
aberrations. Spherical and longitudinal chromatic aberrations will cause
the signal to be rejected by the confocal pinhole. This will get worse
with depth.

If you are going to spend lots of $$$ on a lens, why compromise
performance by using a lens from a different manufacturer which may do
the chromatic aberration correction differently. You may end up with no
correction or too much.

With any optical zoom system it is the NA that is important. This will
limit the resolution. Two similar lenses with same NA . Eg 20 x, zoom 2
NA=1 compared with 40 x, zoom 1 NA=1
Should have identical performance, provided everything else is the same.

So why would you buy the 40x when the 20 x will do the job of both these
objectives?

Of course you need to compare many other things, such as WD, flatness,
chromatic aberration correction etc etc.

I hope that makes more sense.

Steve

Stephen H. Cody

On 05/03/2011, at 10:45 AM, "G. Esteban Fernandez"
<[hidden email]> wrote:

> *****
> To join, leave or search the confocal microscopy listserv, go to:
> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> *****
>
> Steve,
>
> Just to clarify, you think the loss of resolution/signal is more with
> chromatic aberration than by reducing the NA by ~0.2 plus air/water RI
> mismatch?
>
> Thanks,
> Esteban
>
> On Fri, Mar 4, 2011 at 2:46 PM, Stephen Cody <[hidden email]>

wrote:
>
>> *****
>> To join, leave or search the confocal microscopy listserv, go to:
>> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
>> *****
>>
>> G'day Aleksandr,
>>
>> Do you require this lens for confocal and or multiphoton microscopy?
If so
>> you should follow George's advice.
>>
>> If you purchase a high NA 20x (or 25x) lens. Then this will perform
>> perfectly at zoom 1 with a wide field of view, and as described be
George
>> zoom in 2 or 3 times and have the equivalent performance of a 40 or
60x.
>> Saving you the cost of several lenses.
>>
>> If however if you need a 40x for conventional microscopy, then you
may need
>> a 40x.
>>
>> Mismatching objectives from manufactures may be possible, but is not
a good
>> idea, especially if you want to use confocal microscopy to optically
section
>> deep within tissue. The loss of chromatic correction I think would
result in
>> very significant loss of signal strength as you go deeper into tissue
with
>> confocal. Maybe not so problematic if using for conventional
microscopy or
>> multiphoton with NDDs.
>>
>> I wouldn't mix manufactures lenses on confocal if you were hoping for
good
>> depth penetration. Which is likely the reason why you want a water
immersion
>> lens in the first place.
>>
>> Cheers Steve
>>
>> Stephen H. Cody
>>
>> On 26/02/2011, at 11:43 AM, George McNamara
<[hidden email]>
>> wrote:
>>
>>> *****
>>> To join, leave or search the confocal microscopy listserv, go to:
>>> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
>>> *****
>>>
>>> Leica has a 20x/1.0 NA objective lens, and, if I recall correctly, a
>> 25x/0.95NA - if this is on a laser scanning confocal, zoom these
lenses to

>> 2x. NA is what matters.
>>>
>>> On 2/25/2011 10:10 AM, Jurkevic, Aleksandr wrote:
>>>> *****
>>>> To join, leave or search the confocal microscopy listserv, go to:
>>>> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
>>>> *****
>>>>
>>>> Hello,
>>>>
>>>> Leica doesn't currently have a 40x water immersion objective

although I
>> heard that they have one in the pipeline. Has anyone used a 40x water
from a

>> different manufacturer with success on this frame?
>>>>
>>>>
>>>> Aleksandr Jurkevic, PhD
>>>> Associate Director
>>>> Molecular Cytology Core
>>>> University of Missouri-Columbia
>>>> 120 Life Science Center
>>>> 1201 E. Rollins St.
>>>> Columbia, MO 65211-7310
>>>>
>>>>
>>>
>>>
>>> --
>>>
>>>
>>> George McNamara, PhD
>>> Analytical Imaging Core Facility
>>> University of Miami
>>

________________________________

No virus found in this message.
Checked by AVG - www.avg.com
Version: 10.0.1204 / Virus Database: 1435/3482 - Release Date: 03/04/11

[Jerry (Gerald) Sedgewick]

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

Seats are still open for the Scientific Imaging with Photoshop (also
Image J, GIMP, Bridge) and Quantization Seminar in the Washington D.C.
area (Bethesda, MD), sponsored by the Histochemical Society.

Acquisition and post-processing of scientific images are increasingly
being scrutinized, and so knowledge of best practices in imaging is
becoming more crucial.  It is also critical to clearly show visual data
in publication and presentation: learn how to conform image tones to
outputs so that salient features can be seen.

Here is the information on the seminars:

"Scientific imaging: acquisition, post-processing (Photoshop, ImageJ,
Bridge, etc.), correcting for outputs and quantization" series of
seminars are being held March 21 - 25, 2011, in Bethesda, Maryland
(Washington D.C. area) at the Hyatt Regency Hotel (One Bethesda Metro
Center) near the NIH campus on the GREEN line of the Metrotrain.

These seminars will be given by Jerry Sedgewick (author of "Scientific
Imaging with Photoshop: Methods, Measurement and Output," past core
microscopy facility director at University of Minnesota, consultant for
quantization, including Computer Aided Diagnosis from fundus images) and
will focus primarily on scientific images post-processed in Photoshop,
Image J, and related programs using best methods/practices for image
integrity.

Please go to http://www.imagingandanalysis.com/seminars.html for more
information.

Whether you've used Photoshop and related programs in the past, or you
have limited experience, the combination of an in-depth, hands-on
approach and expert guidance make the seminars appropriate for all but
advanced users.  Don't assume your students and staff use it correctly!

Overview

Here is the outline for the seminars:
http://www.imagingandanalysis.com/course.html.

The 3-Day Seminar is an energetic hands on lecture and practice session.
Take this seminar to really implant acquisition, post-processing,
outputting and quantization techniques ($690.00).

Or sign up for the 2-day portion ($450) or the 1-Day Quantization
portion ($250).

I will be available for looking at your images for a free consultation
on quantization or imaging procedures on Saturday, March 26 at the Hyatt
Regency Hotel near the NIH campus. Please email me to let me know
whether you or a member of your lab is coming.

Sign up for seminars at http://imagingandanalysis.com/seminars.html.

--
IMAGING: Image Integrity, Quantitation, Digital Imaging Instruction

Jerry Sedgewick
Sedgewick Initiatives
965 Cromwell Avenue
Saint Paul, MN  55114
651-788-2261

[hidden email]
[hidden email]

http://www.imagingandanalysis.com
http://www.quickphotoshop.com

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

[Naomi Book]

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

Dear List,

I would like to know if any one is currently using a SIM Nikon microscope,
any information regarding the system would be appreciated.

Naomi Book


Naomi Melamed-Book, Ph.D
Bio-Imaging Unit
Life Science Institute
Hebrew University, Givat Ram,
Jerusalem, Israel
phone: [phone]
Fax:[phone]
[hidden email]

 
 

-----Original Message-----
From: Confocal Microscopy List [mailto:[hidden email]] On
Behalf Of Jerry (Gerald) Sedgewick
Sent: Monday, March 07, 2011 6:59 PM
To: [hidden email]
Subject: FINAL REMINDER: PHOTOSHOP AND QUANTIZATION SEMINAR, BETHESDA, MD

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

Seats are still open for the Scientific Imaging with Photoshop (also
Image J, GIMP, Bridge) and Quantization Seminar in the Washington D.C.
area (Bethesda, MD), sponsored by the Histochemical Society.

Acquisition and post-processing of scientific images are increasingly
being scrutinized, and so knowledge of best practices in imaging is
becoming more crucial.  It is also critical to clearly show visual data
in publication and presentation: learn how to conform image tones to
outputs so that salient features can be seen.

Here is the information on the seminars:

"Scientific imaging: acquisition, post-processing (Photoshop, ImageJ,
Bridge, etc.), correcting for outputs and quantization" series of
seminars are being held March 21 - 25, 2011, in Bethesda, Maryland
(Washington D.C. area) at the Hyatt Regency Hotel (One Bethesda Metro
Center) near the NIH campus on the GREEN line of the Metrotrain.

These seminars will be given by Jerry Sedgewick (author of "Scientific
Imaging with Photoshop: Methods, Measurement and Output," past core
microscopy facility director at University of Minnesota, consultant for
quantization, including Computer Aided Diagnosis from fundus images) and
will focus primarily on scientific images post-processed in Photoshop,
Image J, and related programs using best methods/practices for image
integrity.

Please go to http://www.imagingandanalysis.com/seminars.html for more
information.

Whether you've used Photoshop and related programs in the past, or you
have limited experience, the combination of an in-depth, hands-on
approach and expert guidance make the seminars appropriate for all but
advanced users.  Don't assume your students and staff use it correctly!

Overview

Here is the outline for the seminars:
http://www.imagingandanalysis.com/course.html.

The 3-Day Seminar is an energetic hands on lecture and practice session.
Take this seminar to really implant acquisition, post-processing,
outputting and quantization techniques ($690.00).

Or sign up for the 2-day portion ($450) or the 1-Day Quantization
portion ($250).

I will be available for looking at your images for a free consultation
on quantization or imaging procedures on Saturday, March 26 at the Hyatt
Regency Hotel near the NIH campus. Please email me to let me know
whether you or a member of your lab is coming.

Sign up for seminars at http://imagingandanalysis.com/seminars.html.

--
IMAGING: Image Integrity, Quantitation, Digital Imaging Instruction

Jerry Sedgewick
Sedgewick Initiatives
965 Cromwell Avenue
Saint Paul, MN  55114
651-788-2261

[hidden email]
[hidden email]

http://www.imagingandanalysis.com
http://www.quickphotoshop.com

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

[Valeria Berno-2]

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

Hi Aleksander

I actually have one Leica 40X immersion objective. It could be quite old
(it was alreay here when I arrived).

It is a HCX APO L40X/0.80 W U-V-I

why don't you ask for this one?

best

Valeria

On 04/03/2011 22:46, Stephen Cody wrote:

> *****
> To join, leave or search the confocal microscopy listserv, go to:
> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> *****
>
> G'day Aleksandr,
>
> Do you require this lens for confocal and or multiphoton microscopy? If so you should follow George's advice.
>
> If you purchase a high NA 20x (or 25x) lens. Then this will perform perfectly at zoom 1 with a wide field of view, and as described be George zoom in 2 or 3 times and have the equivalent performance of a 40 or 60x. Saving you the cost of several lenses.
>
> If however if you need a 40x for conventional microscopy, then you may need a 40x.
>
> Mismatching objectives from manufactures may be possible, but is not a good idea, especially if you want to use confocal microscopy to optically section deep within tissue. The loss of chromatic correction I think would result in very significant loss of signal strength as you go deeper into tissue with confocal. Maybe not so problematic if using for conventional microscopy or multiphoton with NDDs.
>
> I wouldn't mix manufactures lenses on confocal if you were hoping for good depth penetration. Which is likely the reason why you want a water immersion lens in the first place.
>
> Cheers Steve
>
> Stephen H. Cody
>
> On 26/02/2011, at 11:43 AM, George McNamara<[hidden email]>  wrote:
>
>> *****
>> To join, leave or search the confocal microscopy listserv, go to:
>> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
>> *****
>>
>> Leica has a 20x/1.0 NA objective lens, and, if I recall correctly, a 25x/0.95NA - if this is on a laser scanning confocal, zoom these lenses to 2x. NA is what matters.
>>
>> On 2/25/2011 10:10 AM, Jurkevic, Aleksandr wrote:
>>> *****
>>> To join, leave or search the confocal microscopy listserv, go to:
>>> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
>>> *****
>>>
>>> Hello,
>>>
>>> Leica doesn't currently have a 40x water immersion objective although I heard that they have one in the pipeline. Has anyone used a 40x water from a different manufacturer with success on this frame?
>>>
>>>
>>> Aleksandr Jurkevic, PhD
>>> Associate Director
>>> Molecular Cytology Core
>>> University of Missouri-Columbia
>>> 120 Life Science Center
>>> 1201 E. Rollins St.
>>> Columbia, MO 65211-7310
>>>
>>>
>>
>> --
>>
>>
>> George McNamara, PhD
>> Analytical Imaging Core Facility
>> University of Miami
>

--
Valeria Berno,PhD
Microscopy Facility Manager
MRC Centre for Regenerative Medicine
Institute for Stem Cell Research
University of Edinburgh
Roger Land Building
West Mains Road
Edinburgh
EH9 3JQ
Rm 273
Tel Office 0131 6517265



The University of Edinburgh is a charitable body, registered in
Scotland, with registration number SC005336.

[rjpalmer]

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http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
*****

The Leica global website currently lists 10x, 20x, 40x and 63 x (the
latter NA 0.9) as available objectives.  Perhaps the reality is
different, but these objectives were quite popular and I think it
would have been a bad marketing decision to eliminate any of them.

>*****
>To join, leave or search the confocal microscopy listserv, go to:
>http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
>*****
>
>Hi Aleksander
>
>I actually have one Leica 40X immersion objective. It could be quite old
>(it was alreay here when I arrived).
>
>It is a HCX APO L40X/0.80 W U-V-I
>
>why don't you ask for this one?
>
>best
>
>Valeria
>
>On 04/03/2011 22:46, Stephen Cody wrote:
>>  *****
>>  To join, leave or search the confocal microscopy listserv, go to:
>>  http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
>>  *****
>>
>>  G'day Aleksandr,
>>
>>  Do you require this lens for confocal and or multiphoton
>>microscopy? If so you should follow George's advice.
>>
>>  If you purchase a high NA 20x (or 25x) lens. Then this will
>>perform perfectly at zoom 1 with a wide field of view, and as
>>described be George zoom in 2 or 3 times and have the equivalent
>>performance of a 40 or 60x. Saving you the cost of several lenses.
>>
>>  If however if you need a 40x for conventional microscopy, then you
>>may need a 40x.
>>
>>  Mismatching objectives from manufactures may be possible, but is
>>not a good idea, especially if you want to use confocal microscopy
>>to optically section deep within tissue. The loss of chromatic
>>correction I think would result in very significant loss of signal
>>strength as you go deeper into tissue with confocal. Maybe not so
>>problematic if using for conventional microscopy or multiphoton
>>with NDDs.
>>
>>  I wouldn't mix manufactures lenses on confocal if you were hoping
>>for good depth penetration. Which is likely the reason why you want
>>a water immersion lens in the first place.
>>
>>  Cheers Steve
>>
>>  Stephen H. Cody
>>
>>  On 26/02/2011, at 11:43 AM, George
>>McNamara<[hidden email]>  wrote:
>>
>>>  *****
>>>  To join, leave or search the confocal microscopy listserv, go to:
>>>  http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
>>>  *****
>>>
>>>  Leica has a 20x/1.0 NA objective lens, and, if I recall
>>>correctly, a 25x/0.95NA - if this is on a laser scanning confocal,
>>>zoom these lenses to 2x. NA is what matters.
>>>
>>>  On 2/25/2011 10:10 AM, Jurkevic, Aleksandr wrote:
>>>>  *****
>>>>  To join, leave or search the confocal microscopy listserv, go to:
>>>>  http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
>>>>  *****
>>>>
>>>>  Hello,
>>>>
>>>>  Leica doesn't currently have a 40x water immersion objective
>>>>although I heard that they have one in the pipeline. Has anyone
>>>>used a 40x water from a different manufacturer with success on
>>>>this frame?
>>>>
>>>>
>>>>  Aleksandr Jurkevic, PhD
>>>>  Associate Director
>>>>  Molecular Cytology Core
>>>>  University of Missouri-Columbia
>>>>  120 Life Science Center
>>>>  1201 E. Rollins St.
>>>>  Columbia, MO 65211-7310
>>>>
>>>>
>>>
>>>  --
>>>
>>>
>>>  George McNamara, PhD
>>>  Analytical Imaging Core Facility
>>>  University of Miami
>>
>
>--
>Valeria Berno,PhD
>Microscopy Facility Manager
>MRC Centre for Regenerative Medicine
>Institute for Stem Cell Research
>University of Edinburgh
>Roger Land Building
>West Mains Road
>Edinburgh
>EH9 3JQ
>Rm 273
>Tel Office 0131 6517265
>
>
>
>The University of Edinburgh is a charitable body, registered in
>Scotland, with registration number SC005336.

--
Robert J. Palmer Jr., Ph.D.
Natl Inst Dental Craniofacial Res - Natl Insts Health
Oral Infection and Immunity Branch
Bldg 30, Room 310
30 Convent Drive
Bethesda MD 20892
ph 301-594-0025
fax [phone]

[Phillips, Thomas E.]

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

Leica has just announced that they now have a coverslip corrected 40x water immersion objective.

15506341 HCX PL APO 40x/1.10 W CORR CS
HCX PL APO 40x/1.10 water immersion objective with correction collar optimized for
confocal scanning applications. High transmission from 360-1100 nm, free working
distance: 0.63 mm. For 405 nm excitation correction lens 155063414 has to be used.



Thomas E. Phillips, Ph.D
Professor of Biological Sciences
Director, Molecular Cytology Core
2 Tucker Hall
University of Missouri
Columbia, MO 65211-7400
573-882-4712 (office)
573-882-0123 (fax)
[hidden email]

http://www.biology.missouri.edu/faculty/phillips.html
http://www.biotech.missouri.edu/mcc/

-----Original Message-----
From: Confocal Microscopy List [mailto:[hidden email]] On Behalf Of Robert J. Palmer Jr.
Sent: Wednesday, March 09, 2011 7:42 AM
To: [hidden email]
Subject: Re: 40x water immersion objective on Leica miscroscope

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

The Leica global website currently lists 10x, 20x, 40x and 63 x (the
latter NA 0.9) as available objectives.  Perhaps the reality is
different, but these objectives were quite popular and I think it
would have been a bad marketing decision to eliminate any of them.

>*****
>To join, leave or search the confocal microscopy listserv, go to:
>http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
>*****
>
>Hi Aleksander
>
>I actually have one Leica 40X immersion objective. It could be quite old
>(it was alreay here when I arrived).
>
>It is a HCX APO L40X/0.80 W U-V-I
>
>why don't you ask for this one?
>
>best
>
>Valeria
>
>On 04/03/2011 22:46, Stephen Cody wrote:
>>  *****
>>  To join, leave or search the confocal microscopy listserv, go to:
>>  http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
>>  *****
>>
>>  G'day Aleksandr,
>>
>>  Do you require this lens for confocal and or multiphoton
>>microscopy? If so you should follow George's advice.
>>
>>  If you purchase a high NA 20x (or 25x) lens. Then this will
>>perform perfectly at zoom 1 with a wide field of view, and as
>>described be George zoom in 2 or 3 times and have the equivalent
>>performance of a 40 or 60x. Saving you the cost of several lenses.
>>
>>  If however if you need a 40x for conventional microscopy, then you
>>may need a 40x.
>>
>>  Mismatching objectives from manufactures may be possible, but is
>>not a good idea, especially if you want to use confocal microscopy
>>to optically section deep within tissue. The loss of chromatic
>>correction I think would result in very significant loss of signal
>>strength as you go deeper into tissue with confocal. Maybe not so
>>problematic if using for conventional microscopy or multiphoton
>>with NDDs.
>>
>>  I wouldn't mix manufactures lenses on confocal if you were hoping
>>for good depth penetration. Which is likely the reason why you want
>>a water immersion lens in the first place.
>>
>>  Cheers Steve
>>
>>  Stephen H. Cody
>>
>>  On 26/02/2011, at 11:43 AM, George
>>McNamara<[hidden email]>  wrote:
>>
>>>  *****
>>>  To join, leave or search the confocal microscopy listserv, go to:
>>>  http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
>>>  *****
>>>
>>>  Leica has a 20x/1.0 NA objective lens, and, if I recall
>>>correctly, a 25x/0.95NA - if this is on a laser scanning confocal,
>>>zoom these lenses to 2x. NA is what matters.
>>>
>>>  On 2/25/2011 10:10 AM, Jurkevic, Aleksandr wrote:
>>>>  *****
>>>>  To join, leave or search the confocal microscopy listserv, go to:
>>>>  http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
>>>>  *****
>>>>
>>>>  Hello,
>>>>
>>>>  Leica doesn't currently have a 40x water immersion objective
>>>>although I heard that they have one in the pipeline. Has anyone
>>>>used a 40x water from a different manufacturer with success on
>>>>this frame?
>>>>
>>>>
>>>>  Aleksandr Jurkevic, PhD
>>>>  Associate Director
>>>>  Molecular Cytology Core
>>>>  University of Missouri-Columbia
>>>>  120 Life Science Center
>>>>  1201 E. Rollins St.
>>>>  Columbia, MO 65211-7310
>>>>
>>>>
>>>
>>>  --
>>>
>>>
>>>  George McNamara, PhD
>>>  Analytical Imaging Core Facility
>>>  University of Miami
>>
>
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