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I have that lens; I'm going to bug my Nikon rep about the cap right away!
Craig
On Thu, Feb 21, 2013 at 1:54 AM, Horn Thomas <[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
> *****
>
> Hello Adam,
> I have just received from Nikon an immersion fluid replacement system for
> their water objectives. It consists of objective-dedicated brass caps
> connected by tubing to a syringe pump. However, I have not tried it and I
> do not know if they offer it for your 25 x. Maybe worth contacting Nikon on
> that....
> Best regards,
> Thomas.
>
>
> Dr. Thomas Horn,
> The Single Cell Unit, U1.46
> Department of Biosystems Science and Engineering (D-BSSE)
> Swiss Federal Institute of Technology Zurich (ETH)
> Mattenstrasse 26
> CH 4048 Basel
> Switzerland
> Phone: +41 61 387 3373
> mail: [log in to unmask]
>
>
> -----Original Message-----
> From: Confocal Microscopy List [mailto:[log in to unmask]]
> On Behalf Of Watkins, Simon C
> Sent: 20 February 2013 18:18
> To: [log in to unmask]
> Subject: Re: Long term water immersion imaging
>
> *****
> To join, leave or search the confocal microscopy listserv, go to:
> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> *****
>
> I guess my last comment on the use of Genteal was a little cursory. This
> is a water based gel that was developed for folks with chronic dry eyes.
> It has the same RI as water and works extremely well for long term
> experiments with the new objectives (at least Oly and Nikon). It does work
> better when the sample is mounted in a humidified chamber and allows the
> full working distance of the lens to be used which generally will not work
> with the water RI oils which simply do not generate sufficient surface
> tension. The best thing about Genteal, is that its an over the counter
> medication and available from Amazon (see my earlier post) which means it
> works, and is in-expensive. Working time for the stuff is about 24 hours
> with a humidified chamber. This also assumes the stage is being moved
> around to set positions over time, which is very difficult to do with oil
> As a last comment, as Genteal is water based, cleaning etc is a doddle
> which can be a problem with the water RI oils.
>
>
> Simon Watkins Ph.D
>
> Professor and Vice Chair Cell Biology
> Professor Immunology
> Director Center for Biologic Imaging
> University of Pittsburgh
> Bsts 225 3550 terrace st
> Pittsburgh PA 15261
> Www.cbi.pitt.edu <http://Www.cbi.pitt.edu/>
> 412-352-2277
>
>
>
>
>
>
> On 2/20/13 11:40 AM, "Armstrong, Brian" <[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
> >*****
> >
> >Hello, I think the problem is that these LWD 2P lenses are huge and
> >need a large volume of water for the long working distance. The Zeiss
> >immersion media will not hold surface tension in that volume (already
> >mentioned [although works great for lenses such as 63x/1.2W]). What we
> >did was purchase a variety of rubber washers in a "plumbing kit", the
> >cost was a few dollars. We glued a rubber washer to the slide
> >(silicone, VALAP, cyanoacrylate will all work here) and filled it with
> >water. We imaged with this configuration over several days. A few years
> >back I believe there was a thread on this listserve about using condoms
> >as water dams for this purpose. The size of the condom will depend on
> >the size of the objective. You can cut the condom to suit your needs.
> >Cheers,
> >
> >Brian D Armstrong PhD
> >
> >-----Original Message-----
> >From: Confocal Microscopy List
> >[mailto:[log in to unmask]]
> >On Behalf Of Dmitry Sokolov
> >Sent: Tuesday, February 19, 2013 8:53 PM
> >To: [log in to unmask]
> >Subject: Re: Long term water immersion imaging
> >
> >*****
> >To join, leave or search the confocal microscopy listserv, go to:
> >http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> >*****
> >
> >Hi Ammasi,
> >
> >sorry, I probably missed how the immersion oil was applied: on the top
> >of water or on the top of sample:
> >http://confocal-manawatu.pbworks.com/w/page/63806798/Long%20term%20wate
> >r%2
> >0immersion%20imaging
> >
> >Thank you beforehand,
> >Dmitry
> >
> >*Advanced Knowledge Management*
> >for *MICROSCOPY *and *Image Analysis *
> >-----------------------------------------------------------------------
> >-
> >*Dmitry Sokolov*, Ph.D.
> >Mob: *+64 21 063 5382***
> >[log in to unmask] <mailto:[log in to unmask]>
> >
> >20.02.2013 16:19, Periasamy, Ammasi (ap3t) ?????:
> >> *****
> >> To join, leave or search the confocal microscopy listserv, go to:
> >> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> >> *****
> >>
> >> Sorry, I forgot to mention...
> >> This water immersion oil can be used for any commercially available
> >>objective lens.
> >> We are using this for Nikon, Leica and Olympus. It works.
> >> Good luck.
> >> Ammasi
> >>
> >>
> >> -----Original Message-----
> >> From: Confocal Microscopy List
> >>[mailto:[log in to unmask]] On Behalf Of Periasamy,
> >>Ammasi
> >>(ap3t)
> >> Sent: Tuesday, February 19, 2013 10:17 PM
> >> To: [log in to unmask]
> >> Subject: Re: Long term water immersion imaging
> >>
> >> *****
> >> To join, leave or search the confocal microscopy listserv, go to:
> >> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> >> *****
> >>
> >> Dear All
> >> For water immersion lens, yes if you use water, particularly for
> >>2photon imaging, it will evaporate within few minutes. We went through
> >>this for 2p imaging. I thought there should be a medium of refractive
> >>index same as water. I discussed this issue with our Zeiss sales
> >>representative. She came to my office next day and provided valuable
> >>information. The zeiss sells an immersion oil and its refractive index
> >>is same as water. We have been using this for 2photon or Confocal time
> >>lapse imaging (24 hrs) since 2008. No problem and the image quality is
> >>great. I am not sure anyone raised this question before in the list
> >>server. I apologize for missing this kind of question/help. The price
> >>is super high compared to the regular immersion oil. The price is
> >>about $125-150. Here is the part#
> >> 000000-1252-136 Immersion medium "Immersol" W, oiler 20 ml Visit the
> >>Zeiss market place web site to order it.
> >>www.micro-shop.zeiss.com/us/us_en Hope this helps.
> >> Best wishes,
> >> Ammasi
> >>
> >>
> >> Dr. Ammasi Periasamy
> >> Professor & Center Director
> >> W.M. Keck Center for Cellular Imaging (KCCI) (A University Imaging
> >> Center) Biology, University of Virginia Mail or FedEx: 485 McCormick
> >> Rd.
> >> Charlottesville, VA 22904.
> >> Office Location: Physical Life Sciences Building (B005) 90, Geldard
> >> Drive, Charlottesville, VA 22904
> >> Voice: 434-243-7602 (Office); 982-4869 (lab) Fax:434-982-5210;
> >> Email:[log in to unmask] http://www.kcci.virginia.edu/contact/peri.php
> >> ************************
> >> 12th Annual Workshop on FRET Microscopy, March 11-16, 2013
> >> http://www.kcci.virginia.edu/workshop/workshop2013/index.php
> >> *************************
> >>
> >>
> >> -----Original Message-----
> >> From: Confocal Microscopy List
> >>[mailto:[log in to unmask]] On Behalf Of Watkins, Simon
> >>C
> >> Sent: Tuesday, February 19, 2013 10:00 PM
> >> To: [log in to unmask]
> >> Subject: Re: Long term water immersion imaging
> >>
> >> *****
> >> To join, leave or search the confocal microscopy listserv, go to:
> >> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> >> *****
> >>
> >> For us genteal gel
> >>
> >>http://www.amazon.com/GenTeal-Lubricant-Moderate-Severe-Relief/dp/B001
> >>GBI
> >>S7
> >> Y/ref=pd_bxgy_hpc_img_z works very well..we have used it overnight in
> >>some conditions.. Though this was within an humidified chamber
> >>
> >> Simon Watkins Ph.D
> >>
> >> Professor and Vice Chair Cell Biology Professor Immunology Director
> >> Center for Biologic Imaging University of Pittsburgh Bsts 225 3550
> >> terrace st Pittsburgh PA 15261 Www.cbi.pitt.edu
> >> <http://Www.cbi.pitt.edu/>
> >> 412-352-2277
> >>
> >>
> >>
> >>
> >>
> >>
> >> On 2/19/13 6:04 PM, "Benjamin Hibbs" <[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
> >>> *****
> >>>
> >>> Hi Adam,
> >>>
> >>> I'm sure some of the other members of the list have more experience
> >>> than I. However, perhaps you could try making a continual water
> >>> source like the leica system. I know Steve Cody has developed some
> >>> nifty techniques to maintain the immersion water automatically, but
> >>> even a manual top-up every few hours could help you in conjunction
> >>> with your reservoir approach.
> >>>
> >>> Best of luck,
> >>>
> >>> Ben
> >>>
> >>>
> >>> Ben Hibbs
> >>> Platform Support Officer<Advanced Fluorescence Imaging The Melbourne
> >>> Materials Institute (MMI) University of Melbourne, Victoria 3010,
> >>> Australia
> >>> Email:
> >>> [log in to unmask]<mailto:[log in to unmask]>
> >>> Phone: +61 (0)3 9035-7749
> >>>
> >>>
> >>>
> >>>
> >>>
> >>>
> >>>
> >>> On 20/02/2013, at 7:01 AM, Adam White
> >>> <[log in to unmask]<mailto:[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
> >>> *****
> >>>
> >>> I did a search of the archive but I think I have exhausted most of
> >>> the previous suggestions... We are attempting to do some overnight
> >>> live imaging experiments using 2PE microscopy. We are using the
> >>> Nikon Apo LWD 25x/1.1 water immersion objective on an inverted stand
> >>> and our main problem is keeping the immersion medium in place for
> >>> longer than a couple of hours.
> >>> We
> >>> have tried using the Cargille oil with a 1.335 RI but it does not
> >>> have enough viscosity/surface tension to be useful. We have tried
> >>> using ultrasound gel but this dries out over time and only gives us
> >>> about 5 hours of images. I have also tried all manner of different
> >>> sealants/gloves/o-rings to varying degrees of success/reproducibility.
> >>> The best I have found is to use a stretched-out glove finger (plus a
> >>> sealant) and just fill up the resulting "reservoir" with water.
> >>> This has given us enough volume to get about 10 hours worth of
> >>> imaging but is hard to keep consistent. Obviously, my question is
> >>> whether someone has developed or knows of a better system for doing
> >>> this? Perhaps a perfusion system or a more durable glove-finger
> >>> type solution? I have seen the Leica system but this won't fit our
> >>> objective... Any input you have to offer would be most appreciated.
> >>>
> >>> Best,
> >>> Adam
> >>>
> >>>
> >>> Adam B. White, Ph.D.
> >>> Confocal & Specialized Microscopy Shared Resource Herbert Irving
> >>> Comprehensive Cancer Center Columbia University
> >>> 1130 Saint Nicholas Ave, 222A
> >>> New York, NY 10032
> >
> >
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