Zeiss Axiomat & table
Locked
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Available immediately:
The components of a Zeiss inverted modular widefield Axiomat with cameras, bright/darkfield, phase and Nomarski optics and several objectives: 100X and 50 X planapo The Air Suspension table for the Axiomat.
Please contact me immediately if you are interested. Best regards, Sandy, Sandra K. Masur, PhD Associate Dean for Faculty Development http://www.mssm.edu/forfaculty/development President, Women Faculty Group Professor, Ophthalmology Mount Sinai School of Medicine Box 1183 1 Gustave Levy Place New York NY 10029-6574 telephone: 212-241-0089 fax: 212-289-5945 email: [hidden email] |
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Dr. Masur - I don't suppose that you are just trying to
"give" it away are you?
Bill Miller At 01:37 PM 12/4/2008 -0500, you wrote: Available immediately: |
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Can you believe - we are.
The lucky gift recipient is a group at Albert Einstein College of Medicine who responded first to the email. Bingo! On Dec 4, 2008, at 2:09 PM, Bill Miller wrote:
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Robert Peterson-3-3 |
Experience with FV1000 + Precision Plastics Incubation System?
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Hi all,
I'm stepping out of the office for today, but I was wondering if anyone is using the Olympus FVseries with a precision plastics incubation system? I'm having some problems with evaporation and am looking to bounce ideas off someone with a similar setup. A picture of the system is here: http://farm4.static.flickr.com/3018/3019557380_4634d3493e_b.jpg Thanks! Robert |
peterson.vcf (382 bytes) |
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Marla Feller |
Re: Experience with FV1000 + Precision Plastics Incubation System?
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I am imaging GFP using a two-photon (Olympus Fluoview300). I have the
laser tuned at 910 nm. I accidentally found that using my 488 excitation dichroic led to much more excitation than using the 675 excitation dichroic. This doesn't make any sense to me -- can someone tell me where I can find the absorption spectra of these dichroics? Marla On Dec 9, 2008, at 8:09 AM, Robert Peterson wrote: > Hi all, > I'm stepping out of the office for today, but I was wondering if > anyone is using the Olympus FVseries with a precision plastics > incubation system? I'm having some problems with evaporation and am > looking to bounce ideas off someone with a similar setup. > A picture of the system is here: > http://farm4.static.flickr.com/3018/3019557380_4634d3493e_b.jpg > Thanks! > Robert > > <peterson.vcf> |
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Marla,
1) I changed the thread-name, so that your question is not mixed up with the reference of the one you replied to 2) It makes a lot of sense, what you see........ just spend a second on thinking about the wavelength of your expected emission (!) - then ask what the 675-dichroic will do to that ;-) (...it will only deviate wavelength above 675 to your detectors....). Cheers, Christian Marla Feller schrieb: > I am imaging GFP using a two-photon (Olympus Fluoview300). I have the > laser tuned at 910 nm. I accidentally found that using my 488 > excitation dichroic led to much more excitation than using the 675 > excitation dichroic. This doesn't make any sense to me -- can > someone tell me where I can find the absorption spectra of these > dichroics? > > -- ...................................................................... Dr. Christian M. Müller Clinical Neuroanatomy NeuroScience Center, Hs. 89 Goethe University Frankfurt Heinrich-Hoffmann-Str. 7 D-60528 Frankfurt/M., Germany |
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=
I suppose Marla is using a short pass 675, designed to reflect IR excitation light to the sample, and pass the shorter wavelengths (such as 530 nm ) to the detectors. It should work just fine, as it is specifically designed for 2P use. We have seen similar things on our Zeiss LSM 510 confocal, where certain dichroics work better than others, sometimes unexpectedly. I guess it could be an alignment issue. I am not familiar with the Fluoview, but on the Zeiss, pinholes have to be aligned for any particular light path. Your 655 light path may not have been properly aligned. Otherwise, you may want to ask Olympus who provides their filters and/or ask them for their specs. A 488 mirror is primarily designed to reflect the 488 line, but may also reflect other bands... traditionally, there would be no reason for a 488 mirror to transmit wavelengths around 900 nm (IR), and therefore it is not inconceivable it would work for 2P excitation.... Another (trivial ) reason is that your 488 is actually a 488/ NIR dichroic. We have such filters on our Zeiss LSM too, for instance for simultaneous 2P/VIS excitation. -- Julio Vazquez Fred Hutchinson Cancer Research Center 1100 Fairview Ave. N., mailstop DE-512 Seattle, WA 98109-1024 On Dec 9, 2008, at 10:48 AM, Dr. Christian M. Müller wrote:
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Cameron Nowell |
Re: Experience with FV1000 + Precision Plastics Incubation System?
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In reply to this post by Robert Peterson-3-3
Hi Robert,
While i have no direct experince with your specific incubator they are all very similar. I have found that evaporation can be a really problem if the humudity of your chanber drops too low. Now humidifying the whole incubator is not a good idea as you will surely damage some of the microscope components. But you can humidify your gas supply bu bubbling your gas through one or two bottles (the first filled with water and the second empty to collect excess water in the air). How exactly is your setup configured? Its a bit hard to see through the black box you have:) Cheers Cam Cameron J. Nowell Microscpy Manager Centre for Advanced Microscopy Ludwig Insttue for Cancer Research PO Box 2008 Royal Melbourne Hospital Victoria, 3050 AUSTRALIA Office: +61 3 9341 3155 Mobile: +61422882700 Fax: +61 3 9341 3104 <http://www.ludwig.edu.au/branch/research/platform/microscopy.htm> ________________________________ From: Confocal Microscopy List on behalf of Robert Peterson Sent: Wed 10/12/2008 3:09 AM To: [hidden email] Subject: Experience with FV1000 + Precision Plastics Incubation System? Hi all, I'm stepping out of the office for today, but I was wondering if anyone is using the Olympus FVseries with a precision plastics incubation system? I'm having some problems with evaporation and am looking to bounce ideas off someone with a similar setup. A picture of the system is here: http://farm4.static.flickr.com/3018/3019557380_4634d3493e_b.jpg Thanks! Robert |
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In reply to this post by Julio Vazquez
To be clear, I am not stating just that the 2-photon emission goes up but rather that the intensity of the 910 nm light reaching our sample is brighter!
It must be the dichroic or the alignment. I will follow up on those two possibilities. thank-you, Marla On Dec 9, 2008, at 11:19 AM, Julio Vazquez wrote:
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There is a spectrum of a 488nm dichroic in Fig 3.11 of my
book (below), but it
only goes out to 750 nm - it's still transmitting
there. The spectrum is of an
everyday (low cost) FITC filter set from Chroma. But
if your filters are from
Chroma they will certainly make the spectra available to
you - they are very
helpful that way. I'd imagine the other manufacturers
would be, too. So get
the maker and designation of your filter from Olympus and
I'm sure you will
then be able to get the spectrum from the
maker.
Guy
Optical Imaging Techniques in Cell Biology From: Confocal Microscopy List [mailto:[hidden email]] On Behalf Of Marla Feller Sent: Wednesday, 10 December 2008 6:56 AM To: [hidden email] Subject: Re: Marla's question It must be the dichroic or the alignment. I will follow up on those
two possibilities.
thank-you,
Marla
On Dec 9, 2008, at 11:19 AM, Julio Vazquez wrote:
No virus found in this incoming message. No virus found in this outgoing message. |
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In reply to this post by Marla Feller
If, as Julio mentions, the 675 excitation diochroic is supposed to be a
low-pass, I apologize (it's me, who should have spent a second on
thinking and looking up the Fluoview lightpath...).
Trying to be more constructive: to get a fast estimate of the GFP-related transmission of the dichroics you might illuminate widefield with a 505 or 515nm LED (few cents) and test the two dichroics with the same PMT-settings. However, if 910nm light is indeed dimmer at the sample with the 675 your assumptions are certainly the most valid. Sorry, Christian Marla Feller schrieb: To be clear, I am not stating just that the 2-photon emission goes up but rather that the intensity of the 910 nm light reaching our sample is brighter! -- ...................................................................... Dr. Christian M. Müller Clinical Neuroanatomy JWG University Heinrich-Hoffmann-Str. 7, Hs. 89/2 D-60528 Frankfurt/M., Germany |
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Boswell, Carl A - (cboswell) |
Re: Marla's question
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In reply to this post by Marla Feller
A look at the spectrum of dichroics, for instance
on the Semrock website, shows that many mirrors work well for their intended
range, but are surprisingly variable at wavelengths some distance from
there. A mirror may have significant reflective and transmission
properties in the NIR, because they are not intended to function out
there. Moveover, many of the vendor websites that show the spectra for
their filters/mirrors do not present data very far into the IR or UV (often
starting at 400 and stopping at 750nm), so what the device is doing outside that
range is anybody's guess.
Carl
Carl A. Boswell, Ph.D.
Molecular and Cellular Biology University of Arizona 520-954-7053 FAX 520-621-3709
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