Comparison Zeiss 710 vs Nikon A1
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Pedro Camello |
Comparison Zeiss 710 vs Nikon A1
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Hi all,
has anybody in the list compared Nikon A1 vs Zeiss 710. We´re purchasing a spectral micro with a TIRF module and motorization. Any input will be really wellcome (off list if you prefer) A second question, what is the most close to "live cell" sample to make real tests in confocal? I´m going to travel to test a couple of micros, and to carry or prepapre real living cells is rather complicated for us. We´re especially interested in ion (Ca2+) experiments. Thanks -- Dr Pedro J Camello Dpt Physiology Faculty of Veterinary Sciences University of Extremadura 10071 Caceres Spain Ph: 927257000 Extension 51321/51290 Fax:927257110 |
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Roshma Azeem |
Re: Comparison Zeiss 710 vs Nikon A1
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Hi Pedro, Seems you have short listed A1 and 710. Both the systems are released almost the same time. We have little experience in handling the 710 system but sufficiently exposed with A1. I have mentioned my opinion and observation about these two systems earlier that is given below. A1 gives 4X4K image size, whereas 710 gives 6X6K. But do we really need 6KX6K? When we tried taking 6K images, we have experienced photo bleaching in some attempts. A1 has continuously variable zoom up to 1000X but LSM 710 is limited to 50X. However, we were not given proper training about how to use variable zoom up to 1000X. In my view, the 1000X variable zoom is not that important factor. Fortunately, in 710 there is a master pinhole unlike the earlier 510 that takes time for alignment. However, 710 uses conventional rectangular pinhole but A1 has unique hexagonal pinhole resulting better images. We have checked the same sample in both the systems. The spectral detector of LSM 710 has 34 channels with simultaneously acquisition unlike the previous model 510 that was with 4x8=32. However, the spectral step size is 3nm (3x34=102nm wavelength resolution) and it is not flexible like A1 that has 2.5nm, 6nm and 32nm (resulting 2.5x32=80nm, 6x32=192nm and 10x32= 320nm wavelength resolution). A1 has surprisingly effective unmixing efficiency even in the close range. In LSM710 high wavelength range up to 1100 nm is possible for optimized transmission. I am not sure if the A1 has the same capability. In our (little) experience, 710 gives wonderful sensitivity. Their software Zen 2009 is compatible with Vista/Windows 7. Even anisotropy imaging is possible with LSM 710 (do not know about A1). But you need to purchase this module (extra cost). Nikon software is robust has almost all of the regular modules. However, for Zen we need to pay extra for the add on modules. Some of the regular functions are "optional" with Zen. For simultaneous photo activation and imaging, one need to incorporate Duo system (two heads) into LSM 710 that adds up the cost. However, A1R scan head serves this purpose without any cumbersome modifications and the speed and performance is relatively incomparable. Though there are some annoying "noise" is generated while we use the resonant scanner the speed and performance is still impressive. If you are regularly going for live cell imaging and Ca++ imaging, I feel A1 has so many features and it is not bleaching the dyes or induce unexpected phototoxicity. Though, we found 710 has slightly better sensitivity, we often face bleaching and laser induced toxicity problems for the same set of experiments. Eager to know the inputs of other users. No commercial interest. Roshma. On Sun, Apr 11, 2010 at 4:34 PM, Pedro J Camello <[hidden email]> wrote: Hi all, |
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Ana Malathi |
Re: Comparison Zeiss 710 vs Nikon A1
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Sam's Mail |
Intravital Microscopy in a Shared Resource?
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In reply to this post by Roshma Azeem
Good Day,
Contemplating the challenges of running a 2-photon intravital system in a shared resource environment... Could I get a show of hands from those of you who have successfully (or not so successfully) integrated a core facility managed intravital microscopy program? I'd love to hear from those as well who have started an intravital program in their own laboratories (non core) to talk shop. Kindest Regards, -- Samuel A. Connell Director of Light Microscopy Cell & Tissue Imaging Center St. Jude Children's Research Hospital 262 Danny Thomas Place Memphis, TN 38105-3678 Office (901) 595-2536 Cell (901) 603-3162 [hidden email] ________________________________ Email Disclaimer: www.stjude.org/emaildisclaimer |
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Armstrong, Brian |
Re: Intravital Microscopy in a Shared Resource?
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Yes, I think I have accomplished this.
Brian D Armstrong PhD Light Microscopy Core Manager Beckman Research Institute City of Hope Dept of Neuroscience 1450 E Duarte Rd Duarte, CA 91010 626-256-4673 x62872 http://www.cityofhope.org/research/support/Light-Microscopy-Digital-Imag ing/Pages/default.aspx -----Original Message----- From: Confocal Microscopy List [mailto:[hidden email]] On Behalf Of Connell, Samuel Sent: Tuesday, April 13, 2010 7:42 AM To: [hidden email] Subject: Intravital Microscopy in a Shared Resource? Good Day, Contemplating the challenges of running a 2-photon intravital system in a shared resource environment... Could I get a show of hands from those of you who have successfully (or not so successfully) integrated a core facility managed intravital microscopy program? I'd love to hear from those as well who have started an intravital program in their own laboratories (non core) to talk shop. Kindest Regards, -- Samuel A. Connell Director of Light Microscopy Cell & Tissue Imaging Center St. Jude Children's Research Hospital 262 Danny Thomas Place Memphis, TN 38105-3678 Office (901) 595-2536 Cell (901) 603-3162 [hidden email] ________________________________ Email Disclaimer: www.stjude.org/emaildisclaimer --------------------------------------------------------------------- SECURITY/CONFIDENTIALITY WARNING: This message and any attachments are intended solely for the individual or entity to which they are addressed. This communication may contain information that is privileged, confidential, or exempt from disclosure under applicable law (e.g., personal health information, research data, financial information). Because this e-mail has been sent without encryption, individuals other than the intended recipient may be able to view the information, forward it to others or tamper with the information without the knowledge or consent of the sender. If you are not the intended recipient, or the employee or person responsible for delivering the message to the intended recipient, any dissemination, distribution or copying of the communication is strictly prohibited. If you received the communication in error, please notify the sender immediately by replying to this message and deleting the message and any accompanying files from your system. If, due to the security risks, you do not wish to receive further communications via e-mail, please reply to this message and inform the sender that you do not wish to receive further e-mail from the sender. --------------------------------------------------------------------- |
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Cameron, Lisa |
Re: Intravital Microscopy in a Shared Resource?
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In reply to this post by Sam's Mail
Hello -
I am setting up a Zeiss 710 NLO with Coherent Chameleon Vision-II in a core facility. Installation was recent and am now working with a few main users to get system set up for intravital imaging. So it's a work in progress... Lisa --------------------------------------- Lisa Cameron, Ph.D. Director of Confocal and Light Microscopy Dana Farber Cancer Institute 44 Binney St.; JF 621 Boston, MA 02115 Office phone: 617-582-8824 Fax: 617-582-8750 -----Original Message----- From: Confocal Microscopy List [mailto:[hidden email]] On Behalf Of Connell, Samuel Sent: Tuesday, April 13, 2010 10:42 AM To: [hidden email] Subject: [CONFOCALMICROSCOPY] Intravital Microscopy in a Shared Resource? Good Day, Contemplating the challenges of running a 2-photon intravital system in a shared resource environment... Could I get a show of hands from those of you who have successfully (or not so successfully) integrated a core facility managed intravital microscopy program? I'd love to hear from those as well who have started an intravital program in their own laboratories (non core) to talk shop. Kindest Regards, -- Samuel A. Connell Director of Light Microscopy Cell & Tissue Imaging Center St. Jude Children's Research Hospital 262 Danny Thomas Place Memphis, TN 38105-3678 Office (901) 595-2536 Cell (901) 603-3162 [hidden email] ________________________________ Email Disclaimer: www.stjude.org/emaildisclaimer The information in this e-mail is intended only for the person to whom it is addressed. If you believe this e-mail was sent to you in error and the e-mail contains patient information, please contact the Partners Compliance HelpLine at http://www.partners.org/complianceline . If the e-mail was sent to you in error but does not contain patient information, please contact the sender and properly dispose of the e-mail. |
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Garfield, Susan (NIH/NCI) [E] |
Re: Intravital Microscopy in a Shared Resource?
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In reply to this post by Sam's Mail
I would be interested in this discussion as we are going in this direction, but I haven't figured out how to address animal protocols etc.
============================================= Susan Garfield Facility Head CCR Confocal Microscopy Core Facility Laboratory of Experimental Carcinogenesis Building 37, Room 4134C (office), Room 4137 (lab) NCI, NIH Bethesda, MD 20892 -----Original Message----- From: Connell, Samuel [mailto:[hidden email]] Sent: Tuesday, April 13, 2010 10:42 AM To: [hidden email] Subject: Intravital Microscopy in a Shared Resource? Good Day, Contemplating the challenges of running a 2-photon intravital system in a shared resource environment... Could I get a show of hands from those of you who have successfully (or not so successfully) integrated a core facility managed intravital microscopy program? I'd love to hear from those as well who have started an intravital program in their own laboratories (non core) to talk shop. Kindest Regards, -- Samuel A. Connell Director of Light Microscopy Cell & Tissue Imaging Center St. Jude Children's Research Hospital 262 Danny Thomas Place Memphis, TN 38105-3678 Office (901) 595-2536 Cell (901) 603-3162 [hidden email] ________________________________ Email Disclaimer: www.stjude.org/emaildisclaimer |
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Haberman, Ann |
Re: Intravital Microscopy in a Shared Resource?
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In reply to this post by Sam's Mail
I have also established a core facility successfully. I would be happy to talk about this offline.
Ann Haberman >Good Day, > >Contemplating the challenges of running a 2-photon intravital system in a shared resource environment... > >Could I get a show of hands from those of you who have successfully (or not so successfully) integrated a core facility managed intravital microscopy program? > >I'd love to hear from those as well who have started an intravital program in their own laboratories (non core) to talk shop. > >Kindest Regards, > >-- >Samuel A. Connell >Director of Light Microscopy >Cell & Tissue Imaging Center >St. Jude Children's Research Hospital >262 Danny Thomas Place >Memphis, TN 38105-3678 >Office (901) 595-2536 >Cell (901) 603-3162 >[hidden email] -- Ann Haberman, PhD In Vivo Imaging Facility, Director Department of Laboratory Medicine Yale University School of Medicine 300 Cedar Street TAC S541 New Haven, CT 06510 203-785-7349 203-785-5415 (fax) [hidden email] |
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Cameron Nowell |
Re: Intravital Microscopy in a Shared Resource?
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In reply to this post by Sam's Mail
Hi Samuel,
We have an Olympus MPE system setup up for intravital imaging in our core facility. It works very well. We have anaesthesia and extraction systems attached. More than happy to answer any questions you might have re setting up, running the experiments, ethics etc. Cheers Cam Cameron J. Nowell Microscopy Manager Centre for Advanced Microscopy Ludwig Institute for Cancer Research PO Box 2008 Royal Melbourne Hospital Victoria, 3050 AUSTRALIA Office: +61 3 9341 3155 Mobile: +61422882700 Fax: +61 3 9341 3104 Facility Website -----Original Message----- From: Confocal Microscopy List [mailto:[hidden email]] On Behalf Of Connell, Samuel Sent: Wednesday, 14 April 2010 12:42 AM To: [hidden email] Subject: Intravital Microscopy in a Shared Resource? Good Day, Contemplating the challenges of running a 2-photon intravital system in a shared resource environment... Could I get a show of hands from those of you who have successfully (or not so successfully) integrated a core facility managed intravital microscopy program? I'd love to hear from those as well who have started an intravital program in their own laboratories (non core) to talk shop. Kindest Regards, -- Samuel A. Connell Director of Light Microscopy Cell & Tissue Imaging Center St. Jude Children's Research Hospital 262 Danny Thomas Place Memphis, TN 38105-3678 Office (901) 595-2536 Cell (901) 603-3162 [hidden email] ________________________________ Email Disclaimer: www.stjude.org/emaildisclaimer |
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George McNamara |
Re: Comparison Zeiss 710 vs Nikon A1 ... full comparison requires matching resolution and scan speed
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In reply to this post by Roshma Azeem
Hi listserv,
Repeating what I posted in a message yesterday:
Comments with respect to the post below... At 03:43 PM 4/11/2010, you wrote: Hi Pedro, GM: With a 100x1.4+ NA objective lens at the LSM710's 0.6x zoom (and perfect specimen, i.e. refractive index matching and/or imaging right at the coverglass ... and coverglass 170.0 um) XY resolution = 0.6 * 405 nm (the laser line) / 1.4 = 173 nm (assuming pinhole 1.0 Airy unit, infinitely bright specimen could put sqrt(2) in the denominator). Nyquist sampling for a 2D image; 173 nm / 3 = 57 nm. (I'm not at the LSM710 so don't have the actual image field of view, but this should be close): For 100x lens and 0.6x zoom, field of view should be about 160 um, so 160 um / 6000 pixels = 0.026 um = 26 nm. So, 6kx6k is oversampling by about a factor of two. For a very stable specimen (photostability and vibration isolation), perfect imaging conditions, lots of time (since probably will need Z-series for best deconvolution) and optimized confocal deconvolution algorithm, the 6kx6k setting might be useful. How much more useful than 4kx4k ... please do the experiment, publish and send me the reference. GM Most confocal microscope software have "optimize" XY resolution buttons - when in doubt, click on it. Incidentally, Paul Goodwin, Applied Precision, told me Biotium CF405 fluorophore is very good for immunofluorescence (context: OMX nanoscope). Might be a good starting point for an experiment such as above. DAPI or Hoechst 33342 or 33358 in a perfect mounting medium could also be a good choice for the above. A1 has continuously variable zoom up to 1000X but LSM 710 is limited to 50X. However, we were not given proper training about how to use variable zoom up to 1000X. In my view, the 1000X variable zoom is not that important factor. GM: Managing both a 4-pinhole LSM510 and a LSM710, I teach users the same thing on both for multi-color fluorescence (ex. "colocalization"): for the longest wavelength emission channel, select 1 Airy button, then match the pinhole for the other channels. This way the (nominal) optical section thickness is matched. In multitrack mode the pinhole setting can be changed between tracks. The spectral detector of LSM 710 has 34 channels with simultaneously acquisition unlike the previous model 510 that was with 4x8=32. However, the spectral step size is 3nm (3x34=102nm wavelength resolution) and it is not flexible like A1 that has 2.5nm, 6nm and 32nm (resulting 2.5x32=80nm, 6x32=192nm and 10x32= 320nm wavelength resolution). A1 has surprisingly effective unmixing efficiency even in the close range. GM: LSM710/ZEN's 32-channel QUASAR detector default is ~10 nm, but is adjustable (5 and 2.5 nm if I recall). When using the flanking PMTs the spectral window for these can be adjusted in 1 nm steps - this is done with two prism/slits - see the LSM710 internals schematic on the Zeiss web site. Going to 1 nm might be useful to play around with the peaks vs off-peaks for Europium or Terbium (which have long fluorescence lifetimes so scan speed would need to be very slow, might be interesting if you have the time to do a spectral scan overnight). In LSM710 high wavelength range up to 1100 nm is possible for optimized transmission. I am not sure if the A1 has the same capability. In our (little) experience, 710 gives wonderful sensitivity. Their software Zen 2009 is compatible with Vista/Windows 7. Even anisotropy imaging is possible with LSM 710 (do not know about A1). But you need to purchase this module (extra cost). GM: See top of message - scan speed, resolution, laser power at specimen, need to be matched to make valid comparison. Eager to know the inputs of other users. George McNamara, Ph.D. Image Core Manager Analytical Imaging Core Facility University of Miami, Miller School of Medicine Miami, FL 33136 [hidden email] [hidden email] 305-243-8436 office http://www.sylvester.org/AICF (Analytical Imaging Core Facility) http://www.sylvester.org/AICF/pubspectra.zip (the entire 2000+ spectra .xlsx file is in the zip file) http://home.earthlink.net/~geomcnamara |
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Andreas Bruckbauer |
Re: Comparison Zeiss 710 vs Nikon A1 ... full comparison requires matching resolution and scan speed
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On the Olympus FV1000 the signal varies (intentionally) with the scan speed, longer dwell time = more photons. Zeiss seems to give out something like counting rates which are then the same for different dwell times. It would be good if the manufacturers give more details.
best wishes Andreas -----Original Message-----
From: George McNamara <[hidden email]> To: [hidden email] Sent: Sun, 9 May 2010 14:58 Subject: Re: Comparison Zeiss 710 vs Nikon A1 ... full comparison requires matching resolution and scan speed
Hi listserv,
Repeating what I posted in a message yesterday:
Comments with respect to the post below... At 03:43 PM 4/11/2010, you wrote: Hi Pedro, GM: With a 100x1.4+ NA objective lens at the LSM710's 0.6x zoom (and perfect specimen, i.e. refractive index matching and/or imaging right at the coverglass ... and coverglass 170.0 um) XY resolution = 0.6 * 405 nm (the laser line) / 1.4 = 173 nm (assuming pinhole 1.0 Airy unit, infinitely bright specimen could put sqrt(2) in the denominator). Nyquist sampling for a 2D image; 173 nm / 3 = 57 nm. (I'm not at the LSM710 so don't have the actual image field of view, but this should be close): For 100x lens and 0.6x zoom, field of view should be about 160 um, so 160 um / 6000 pixels = 0.026 um = 26 nm. So, 6kx6k is oversampling by about a factor of two. For a very stable specimen (photostability and vibration isolation), perfect imaging conditions, lots of time (since probably will need Z-series for best deconvolution) and optimized confocal deconvolution algorithm, the 6kx6k setting might be useful. How much more useful than 4kx4k ... please do the experiment, publish and send me the reference. GM Most confocal microscope software have "optimize" XY resolution buttons - when in doubt, click on it. Incidentally, Paul Goodwin, Applied Precision, told me Biotium CF405 fluorophore is very good for immunofluorescence (context: OMX nanoscope). Might be a good starting point for an experiment such as above. DAPI or Hoechst 33342 or 33358 in a perfect mounting medium could also be a good choice for the above. A1 has continuously variable zoom up to 1000X but LSM 710 is limited to 50X. However, we were not given proper training about how to use variable zoom up to 1000X. In my view, the 1000X variable zoom is not that important factor. GM: Managing both a 4-pinhole LSM510 and a LSM710, I teach users the same thing on both for multi-color fluorescence (ex. "colocalization"): for the longest wavelength emission channel, select 1 Airy button, then match the pinhole for the other channels. This way the (nominal) optical section thickness is matched. In multitrack mode the pinhole setting can be changed between tracks. The spectral detector of LSM 710 has 34 channels with simultaneously acquisition unlike the previous model 510 that was with 4x8=32. However, the spectral step size is 3nm (3x34=102nm wavelength resolution) and it is not flexible like A1 that has 2.5nm, 6nm and 32nm (resulting 2.5x32=80nm, 6x32=192nm and 10x32= 320nm wavelength resolution). A1 has surprisingly effective unmixing efficiency even in the close range. GM: LSM710/ZEN's 32-channel QUASAR detector default is ~10 nm, but is adjustable (5 and 2.5 nm if I recall). When using the flanking PMTs the spectral window for these can be adjusted in 1 nm steps - this is done with two prism/slits - see the LSM710 internals schematic on the Zeiss web site. Going to 1 nm might be useful to play around with the peaks vs off-peaks for Europium or Terbium (which have long fluorescence lifetimes so scan speed would need to be very slow, might be interesting if you have the time to do a spectral scan overnight). In LSM710 high wavelength range up to 1100 nm is possible for optimized transmission. I am not sure if the A1 has the same capability. In our (little) experience, 710 gives wonderful sensitivity. Their software Zen 2009 is compatible with Vista/Windows 7. Even anisotropy imaging is possible with LSM 710 (do not know about A1). But you need to purchase this module (extra cost). GM: See top of message - scan speed, resolution, laser power at specimen, need to be matched to make valid comparison. Eager to know the inputs of other users. George McNamara, Ph.D. Image Core Manager Analytical Imaging Core Facility University of Miami, Miller School of Medicine Miami, FL 33136 [hidden email] [hidden email] 305-243-8436 office http://www.sylvester.org/AICF (Analytical Imaging Core Facility) http://www.sylvester.org/AICF/pubspectra.zip (the entire 2000+ spectra .xlsx file is in the zip file) http://home.earthlink.net/~geomcnamara |
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leoncio vergara |
Re: Comparison Zeiss 710 vs Nikon A1 ... full comparison requires matching resolution and scan speed
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The same is with the Nikon A1R, slower scan speeds result in
more intense images. Which makes sense to me. I have worked with a Zeiss LSM510 for nearly 8 years and I have noticed
that there is probably a compensation factor applied to the images as an
attempt to keep the mean intensity constant. The result are weird effects on
intensity upon changes in scanning speed. I have never been happy with that. I rather prefer the more
intuitive behavior of the Olympus and Nikon systems. Leoncio From: Confocal Microscopy
List [mailto:[hidden email]] On Behalf Of Andreas
Bruckbauer On the Olympus FV1000 the signal varies (intentionally) with the
scan speed, longer dwell time = more photons. Zeiss seems to give out something
like counting rates which are then the same for different dwell times. It would
be good if the manufacturers give more details. -----Original Message----- Hi listserv, I've noticed on both the
Leica SP5, Zeiss LSM510 and Zeiss LSM710 that "faster is brighter"
for many fluorophores. That is, a 0.4 us dwell time results in a brighter image
than a 4 us which is brighter than a 40 us (then repeated 0.4 us, which
resulted in the same brightness as the first 0.4 us dwell time - so not
photobleaching -- not all fluorophores in the specimen [Desmid slide from
Carolina] changed brightness). This implies that <0.4 us might be even
brighter - i.e. resonant scanner mode. A couple of possible explanations (not
mutually exclusive): a) photophysics (possibly
caused by fluorophore-O2 photochemistry) - re TRex (PubMed 19337661), papers by
Sanden/Spielmann/Widergren et al (PubMed 20375039, 20196585, 19007245,
17385841). b) calibration method(s) by
Zeiss and/or Leica that try to (but do not always) match output at all
settings. c) other???
(suggestions/comments welcome!). See also high speed scanning has the potential to increase fluorescence
yield and to reduce photobleaching. Borlinghaus RT. Microsc Res Tech. 2006
Sep;69(9):689-92.PMID: 16878313 Since I can obtain brighter or dimmer fluorescence on the same
field of view on the same microscope system by simply changing scan speed, I
submit that between microscope comparisons are just anecdotes unless every
variable is controlled and reported. Even then there may be variables under the
hood that the confocal manufacturer has not mentioned, such as the possibility
that the system is "calibrated" to all scan speeds output more or
less the same intensity. Hi Pedro,
A1 has continuously variable zoom up to 1000X but LSM 710 is
limited to 50X. However, we were not given proper training about how to use
variable zoom up to 1000X. In my view, the 1000X variable zoom is not that
important factor.
The spectral detector of LSM 710 has 34 channels with
simultaneously acquisition unlike the previous model 510 that was with 4x8=32.
However, the spectral step size is 3nm (3x34=102nm wavelength resolution) and
it is not flexible like A1 that has 2.5nm, 6nm and 32nm (resulting 2.5x32=80nm,
6x32=192nm and 10x32= 320nm wavelength resolution). A1 has surprisingly
effective unmixing efficiency even in the close range.
In LSM710 high wavelength range up to 1100 nm is possible for
optimized transmission. I am not sure if the A1 has the same capability. In our
(little) experience, 710 gives wonderful sensitivity. Their software Zen 2009
is compatible with Vista/Windows 7. Even anisotropy imaging is possible with
LSM 710 (do not know about A1). But you need to purchase this module (extra
cost).
Eager to know the inputs of other users. Hi all, has anybody in the list
compared Nikon A1 vs Zeiss 710. We´re purchasing a spectral micro with a TIRF
module and motorization. Any input will be really wellcome (off list if you prefer) A second question, what is
the most close to "live cell" sample to make real tests in confocal? I´m
going to travel to test a couple of micros, and to carry or prepapre real
living cells is rather complicated for us. We´re especially interested in ion (Ca2+) experiments. Thanks -- Dr Pedro J Camello Dpt Physiology Faculty of Veterinary
Sciences University of Extremadura 10071 Caceres Spain Ph: 927257000 Extension
51321/51290 Fax:927257110
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Glen MacDonald-2 |
Re: Comparison Zeiss 710 vs Nikon A1 ... full comparison requires matching resolution and scan speed
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In reply to this post by Andreas Bruckbauer
The FV-1000 also allows automatic scaling of the HV to maintain PMT sensitivity when switching from fast scanning at 2 µs to full resolution scanning at any longer dwell time. I wonder if the Zeiss has something similar that alters the detectors over all scan rates, but which is less obvious and has a less direct user control than the Fluoview's "Auto HV" button that sits next to the dwell time selector.
Glen On May 10, 2010, at 1:45 PM, Andreas Bruckbauer wrote: > On the Olympus FV1000 the signal varies (intentionally) with the scan speed, longer dwell time = more photons. Zeiss seems to give out something like counting rates which are then the same for different dwell times. It would be good if the manufacturers give more details. > > best wishes > > Andreas > > > > -----Original Message----- > From: George McNamara <[hidden email]> > To: [hidden email] > Sent: Sun, 9 May 2010 14:58 > Subject: Re: Comparison Zeiss 710 vs Nikon A1 ... full comparison requires matching resolution and scan speed > > Hi listserv, > > Repeating what I posted in a message yesterday: > > I've noticed on both the Leica SP5, Zeiss LSM510 and Zeiss LSM710 that "faster is brighter" for many fluorophores. That is, a 0.4 us dwell time results in a brighter image than a 4 us which is brighter than a 40 us (then repeated 0.4 us, which resulted in the same brightness as the first 0.4 us dwell time - so not photobleaching -- not all fluorophores in the specimen [Desmid slide from Carolina] changed brightness). This implies that <0.4 us might be even brighter - i.e. resonant scanner mode. A couple of possible explanations (not mutually exclusive): > a) photophysics (possibly caused by fluorophore-O2 photochemistry) - re TRex (PubMed 19337661), papers by Sanden/Spielmann/Widergren et al (PubMed 20375039, 20196585, 19007245, 17385841). > b) calibration method(s) by Zeiss and/or Leica that try to (but do not always) match output at all settings. > c) other??? (suggestions/comments welcome!). > See also > > high speed scanning has the potential to increase fluorescence yield and to reduce photobleaching. Borlinghaus RT. Microsc Res Tech. 2006 Sep;69(9):689-92.PMID: 16878313 > > Since I can obtain brighter or dimmer fluorescence on the same field of view on the same microscope system by simply changing scan speed, I submit that between microscope comparisons are just anecdotes unless every variable is controlled and reported. Even then there may be variables under the hood that the confocal manufacturer has not mentioned, such as the possibility that the system is "calibrated" to all scan speeds output more or less the same intensity. > > > > Comments with respect to the post below... > > > At 03:43 PM 4/11/2010, you wrote: > >> Hi Pedro, >> >> Seems you have short listed A1 and 710. Both the systems are released almost the same time. We have little experience in handling the 710 system but sufficiently exposed with A1. I have mentioned my opinion and observation about these two systems earlier that is given below. >> >> A1 gives 4X4K image size, whereas 710 gives 6X6K. But do we really need 6KX6K? When we tried taking 6K images, we have experienced photo bleaching in some attempts. > > GM: With a 100x1.4+ NA objective lens at the LSM710's 0.6x zoom (and perfect specimen, i.e. refractive index matching and/or imaging right at the coverglass ... and coverglass 170.0 um) > > XY resolution = 0.6 * 405 nm (the laser line) / 1.4 = 173 nm (assuming pinhole 1.0 Airy unit, infinitely bright specimen could put sqrt(2) in the denominator). > Nyquist sampling for a 2D image; 173 nm / 3 = 57 nm. > > (I'm not at the LSM710 so don't have the actual image field of view, but this should be close): > > For 100x lens and 0.6x zoom, field of view should be about 160 um, so > > 160 um / 6000 pixels = 0.026 um = 26 nm. > > So, 6kx6k is oversampling by about a factor of two. For a very stable specimen (photostability and vibration isolation), perfect imaging conditions, lots of time (since probably will need Z-series for best deconvolution) and optimized confocal deconvolution algorithm, the 6kx6k setting might be useful. How much more useful than 4kx4k ... please do the experiment, publish and send me the reference. > > GM Most confocal microscope software have "optimize" XY resolution buttons - when in doubt, click on it. Incidentally, Paul Goodwin, Applied Precision, told me Biotium CF405 fluorophore is very good for immunofluorescence (context: OMX nanoscope). Might be a good starting point for an experiment such as above. DAPI or Hoechst 33342 or 33358 in a perfect mounting medium could also be a good choice for the above. > >> A1 has continuously variable zoom up to 1000X but LSM 710 is limited to 50X. However, we were not given proper training about how to use variable zoom up to 1000X. In my view, the 1000X variable zoom is not that important factor. >> >> Fortunately, in 710 there is a master pinhole unlike the earlier 510 that takes time for alignment. However, 710 uses conventional rectangular pinhole but A1 has unique hexagonal pinhole resulting better images. We have checked the same sample in both the systems. > > GM: Managing both a 4-pinhole LSM510 and a LSM710, I teach users the same thing on both for multi-color fluorescence (ex. "colocalization"): for the longest wavelength emission channel, select 1 Airy button, then match the pinhole for the other channels. This way the (nominal) optical section thickness is matched. In multitrack mode the pinhole setting can be changed between tracks. > > >> The spectral detector of LSM 710 has 34 channels with simultaneously acquisition unlike the previous model 510 that was with 4x8=32. However, the spectral step size is 3nm (3x34=102nm wavelength resolution) and it is not flexible like A1 that has 2.5nm, 6nm and 32nm (resulting 2.5x32=80nm, 6x32=192nm and 10x32= 320nm wavelength resolution). A1 has surprisingly effective unmixing efficiency even in the close range. > > GM: LSM710/ZEN's 32-channel QUASAR detector default is ~10 nm, but is adjustable (5 and 2.5 nm if I recall). When using the flanking PMTs the spectral window for these can be adjusted in 1 nm steps - this is done with two prism/slits - see the LSM710 internals schematic on the Zeiss web site. Going to 1 nm might be useful to play around with the peaks vs off-peaks for Europium or Terbium (which have long fluorescence lifetimes so scan speed would need to be very slow, might be interesting if you have the time to do a spectral scan overnight). > >> In LSM710 high wavelength range up to 1100 nm is possible for optimized transmission. I am not sure if the A1 has the same capability. In our (little) experience, 710 gives wonderful sensitivity. Their software Zen 2009 is compatible with Vista/Windows 7. Even anisotropy imaging is possible with LSM 710 (do not know about A1). But you need to purchase this module (extra cost). >> >> Nikon software is robust has almost all of the regular modules. However, for Zen we need to pay extra for the add on modules. Some of the regular functions are "optional" with Zen. >> >> For simultaneous photo activation and imaging, one need to incorporate Duo system (two heads) into LSM 710 that adds up the cost. However, A1R scan head serves this purpose without any cumbersome modifications and the speed and performance is relatively incomparable. Though there are some annoying "noise" is generated while we use the resonant scanner the speed and performance is still impressive. >> >> If you are regularly going for live cell imaging and Ca++ imaging, I feel A1 has so many features and it is not bleaching the dyes or induce unexpected phototoxicity. Though, we found 710 has slightly better sensitivity, we often face bleaching and laser induced toxicity problems for the same set of experiments. > > GM: See top of message - scan speed, resolution, laser power at specimen, need to be matched to make valid comparison. > > >> Eager to know the inputs of other users. >> >> No commercial interest. >> >> Roshma. >> >> >> >> On Sun, Apr 11, 2010 at 4:34 PM, Pedro J Camello <[hidden email]> wrote: >> Hi all, >> >> has anybody in the list compared Nikon A1 vs Zeiss 710. We´re purchasing a >> spectral micro with a TIRF module and motorization. Any input will be >> really wellcome (off list if you prefer) >> >> A second question, what is the most close to "live cell" sample to make >> real tests in confocal? I´m going to travel to test a couple of micros, >> and to carry or prepapre real living cells is rather complicated for us. >> We´re especially interested in ion (Ca2+) experiments. >> >> Thanks >> >> >> -- >> Dr Pedro J Camello >> Dpt Physiology >> Faculty of Veterinary Sciences >> University of Extremadura >> 10071 Caceres >> Spain >> Ph: 927257000 Extension 51321/51290 >> Fax:927257110 >> > > > > > > > George McNamara, Ph.D. > Image Core Manager > Analytical Imaging Core Facility > University of Miami, Miller School of Medicine > Miami, FL 33136 > [hidden email] > [hidden email] > 305-243-8436 office > http://www.sylvester.org/AICF (Analytical Imaging Core Facility) > http://www.sylvester.org/AICF/pubspectra.zip (the entire 2000+ spectra .xlsx file is in the zip file) > http://home.earthlink.net/~geomcnamara Glen MacDonald Core for Communication Research Virginia Merrill Bloedel Hearing Research Center Box 357923 University of Washington Seattle, WA 98195-7923 USA (206) 616-4156 [hidden email] |
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Craig Brideau |
Re: Comparison Zeiss 710 vs Nikon A1 ... full comparison requires matching resolution and scan speed
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The Nikon systems I have used allow you to manually select the pixel
dwell time. We use this feature quite often to help clean up images with low photon count. It is somewhat functionally similar to averaging, but has slightly different ramifications to the image information. Also photobleaching must be controlled when using longer dwell times. The good news is when we have bright samples we can set the dwell time to minimum. Craig On Mon, May 10, 2010 at 3:11 PM, Glen MacDonald <[hidden email]> wrote: > The FV-1000 also allows automatic scaling of the HV to maintain PMT sensitivity when switching from fast scanning at 2 µs to full resolution scanning at any longer dwell time. I wonder if the Zeiss has something similar that alters the detectors over all scan rates, but which is less obvious and has a less direct user control than the Fluoview's "Auto HV" button that sits next to the dwell time selector. > > Glen > On May 10, 2010, at 1:45 PM, Andreas Bruckbauer wrote: > >> On the Olympus FV1000 the signal varies (intentionally) with the scan speed, longer dwell time = more photons. Zeiss seems to give out something like counting rates which are then the same for different dwell times. It would be good if the manufacturers give more details. >> >> best wishes >> >> Andreas >> >> >> >> -----Original Message----- >> From: George McNamara <[hidden email]> >> To: [hidden email] >> Sent: Sun, 9 May 2010 14:58 >> Subject: Re: Comparison Zeiss 710 vs Nikon A1 ... full comparison requires matching resolution and scan speed >> >> Hi listserv, >> >> Repeating what I posted in a message yesterday: >> >> I've noticed on both the Leica SP5, Zeiss LSM510 and Zeiss LSM710 that "faster is brighter" for many fluorophores. That is, a 0.4 us dwell time results in a brighter image than a 4 us which is brighter than a 40 us (then repeated 0.4 us, which resulted in the same brightness as the first 0.4 us dwell time - so not photobleaching -- not all fluorophores in the specimen [Desmid slide from Carolina] changed brightness). This implies that <0.4 us might be even brighter - i.e. resonant scanner mode. A couple of possible explanations (not mutually exclusive): >> a) photophysics (possibly caused by fluorophore-O2 photochemistry) - re TRex (PubMed 19337661), papers by Sanden/Spielmann/Widergren et al (PubMed 20375039, 20196585, 19007245, 17385841). >> b) calibration method(s) by Zeiss and/or Leica that try to (but do not always) match output at all settings. >> c) other??? (suggestions/comments welcome!). >> See also >> >> high speed scanning has the potential to increase fluorescence yield and to reduce photobleaching. Borlinghaus RT. Microsc Res Tech. 2006 Sep;69(9):689-92.PMID: 16878313 >> >> Since I can obtain brighter or dimmer fluorescence on the same field of view on the same microscope system by simply changing scan speed, I submit that between microscope comparisons are just anecdotes unless every variable is controlled and reported. Even then there may be variables under the hood that the confocal manufacturer has not mentioned, such as the possibility that the system is "calibrated" to all scan speeds output more or less the same intensity. >> >> >> >> Comments with respect to the post below... >> >> >> At 03:43 PM 4/11/2010, you wrote: >> >>> Hi Pedro, >>> >>> Seems you have short listed A1 and 710. Both the systems are released almost the same time. We have little experience in handling the 710 system but sufficiently exposed with A1. I have mentioned my opinion and observation about these two systems earlier that is given below. >>> >>> A1 gives 4X4K image size, whereas 710 gives 6X6K. But do we really need 6KX6K? When we tried taking 6K images, we have experienced photo bleaching in some attempts. >> >> GM: With a 100x1.4+ NA objective lens at the LSM710's 0.6x zoom (and perfect specimen, i.e. refractive index matching and/or imaging right at the coverglass ... and coverglass 170.0 um) >> >> XY resolution = 0.6 * 405 nm (the laser line) / 1.4 = 173 nm (assuming pinhole 1.0 Airy unit, infinitely bright specimen could put sqrt(2) in the denominator). >> Nyquist sampling for a 2D image; 173 nm / 3 = 57 nm. >> >> (I'm not at the LSM710 so don't have the actual image field of view, but this should be close): >> >> For 100x lens and 0.6x zoom, field of view should be about 160 um, so >> >> 160 um / 6000 pixels = 0.026 um = 26 nm. >> >> So, 6kx6k is oversampling by about a factor of two. For a very stable specimen (photostability and vibration isolation), perfect imaging conditions, lots of time (since probably will need Z-series for best deconvolution) and optimized confocal deconvolution algorithm, the 6kx6k setting might be useful. How much more useful than 4kx4k ... please do the experiment, publish and send me the reference. >> >> GM Most confocal microscope software have "optimize" XY resolution buttons - when in doubt, click on it. Incidentally, Paul Goodwin, Applied Precision, told me Biotium CF405 fluorophore is very good for immunofluorescence (context: OMX nanoscope). Might be a good starting point for an experiment such as above. DAPI or Hoechst 33342 or 33358 in a perfect mounting medium could also be a good choice for the above. >> >>> A1 has continuously variable zoom up to 1000X but LSM 710 is limited to 50X. However, we were not given proper training about how to use variable zoom up to 1000X. In my view, the 1000X variable zoom is not that important factor. >>> >>> Fortunately, in 710 there is a master pinhole unlike the earlier 510 that takes time for alignment. However, 710 uses conventional rectangular pinhole but A1 has unique hexagonal pinhole resulting better images. We have checked the same sample in both the systems. >> >> GM: Managing both a 4-pinhole LSM510 and a LSM710, I teach users the same thing on both for multi-color fluorescence (ex. "colocalization"): for the longest wavelength emission channel, select 1 Airy button, then match the pinhole for the other channels. This way the (nominal) optical section thickness is matched. In multitrack mode the pinhole setting can be changed between tracks. >> >> >>> The spectral detector of LSM 710 has 34 channels with simultaneously acquisition unlike the previous model 510 that was with 4x8=32. However, the spectral step size is 3nm (3x34=102nm wavelength resolution) and it is not flexible like A1 that has 2.5nm, 6nm and 32nm (resulting 2.5x32=80nm, 6x32=192nm and 10x32= 320nm wavelength resolution). A1 has surprisingly effective unmixing efficiency even in the close range. >> >> GM: LSM710/ZEN's 32-channel QUASAR detector default is ~10 nm, but is adjustable (5 and 2.5 nm if I recall). When using the flanking PMTs the spectral window for these can be adjusted in 1 nm steps - this is done with two prism/slits - see the LSM710 internals schematic on the Zeiss web site. Going to 1 nm might be useful to play around with the peaks vs off-peaks for Europium or Terbium (which have long fluorescence lifetimes so scan speed would need to be very slow, might be interesting if you have the time to do a spectral scan overnight). >> >>> In LSM710 high wavelength range up to 1100 nm is possible for optimized transmission. I am not sure if the A1 has the same capability. In our (little) experience, 710 gives wonderful sensitivity. Their software Zen 2009 is compatible with Vista/Windows 7. Even anisotropy imaging is possible with LSM 710 (do not know about A1). But you need to purchase this module (extra cost). >>> >>> Nikon software is robust has almost all of the regular modules. However, for Zen we need to pay extra for the add on modules. Some of the regular functions are "optional" with Zen. >>> >>> For simultaneous photo activation and imaging, one need to incorporate Duo system (two heads) into LSM 710 that adds up the cost. However, A1R scan head serves this purpose without any cumbersome modifications and the speed and performance is relatively incomparable. Though there are some annoying "noise" is generated while we use the resonant scanner the speed and performance is still impressive. >>> >>> If you are regularly going for live cell imaging and Ca++ imaging, I feel A1 has so many features and it is not bleaching the dyes or induce unexpected phototoxicity. Though, we found 710 has slightly better sensitivity, we often face bleaching and laser induced toxicity problems for the same set of experiments. >> >> GM: See top of message - scan speed, resolution, laser power at specimen, need to be matched to make valid comparison. >> >> >>> Eager to know the inputs of other users. >>> >>> No commercial interest. >>> >>> Roshma. >>> >>> >>> >>> On Sun, Apr 11, 2010 at 4:34 PM, Pedro J Camello <[hidden email]> wrote: >>> Hi all, >>> >>> has anybody in the list compared Nikon A1 vs Zeiss 710. We´re purchasing a >>> spectral micro with a TIRF module and motorization. Any input will be >>> really wellcome (off list if you prefer) >>> >>> A second question, what is the most close to "live cell" sample to make >>> real tests in confocal? I´m going to travel to test a couple of micros, >>> and to carry or prepapre real living cells is rather complicated for us. >>> We´re especially interested in ion (Ca2+) experiments. >>> >>> Thanks >>> >>> >>> -- >>> Dr Pedro J Camello >>> Dpt Physiology >>> Faculty of Veterinary Sciences >>> University of Extremadura >>> 10071 Caceres >>> Spain >>> Ph: 927257000 Extension 51321/51290 >>> Fax:927257110 >>> >> >> >> >> >> >> >> George McNamara, Ph.D. >> Image Core Manager >> Analytical Imaging Core Facility >> University of Miami, Miller School of Medicine >> Miami, FL 33136 >> [hidden email] >> [hidden email] >> 305-243-8436 office >> http://www.sylvester.org/AICF (Analytical Imaging Core Facility) >> http://www.sylvester.org/AICF/pubspectra.zip (the entire 2000+ spectra .xlsx file is in the zip file) >> http://home.earthlink.net/~geomcnamara > > > > Glen MacDonald > Core for Communication Research > Virginia Merrill Bloedel Hearing Research Center > Box 357923 > University of Washington > Seattle, WA 98195-7923 USA > (206) 616-4156 > [hidden email] > |
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Stanislav Vitha |
Re: Comparison Zeiss 710 vs Nikon A1 ... full comparison requires matching resolution and scan speed
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In reply to this post by George McNamara
I would expect that in the photon counting (PC) mode all the different
confocals beahve as they should, i.e., longer dwell time = more counts. Since PC is generally less sensitive to detector voltage fluctuations, it is a good idea to use this mode for quantitative measurements anyway. Now, the Olympus FV1000 uses "Hybrid" photon counting. Can somebody please comment what exactly it is? Thanks. Stan On Mon, 10 May 2010 14:11:51 -0700, Glen MacDonald <[hidden email]> wrote: >The FV-1000 also allows automatic scaling of the HV to maintain PMT sensitivity when switching from fast scanning at 2 µs to full resolution scanning at any longer dwell time. I wonder if the Zeiss has something similar that alters the detectors over all scan rates, but which is less obvious and has a less direct user control than the Fluoview's "Auto HV" button that sits next to the dwell time selector. > >Glen >On May 10, 2010, at 1:45 PM, Andreas Bruckbauer wrote: > >> On the Olympus FV1000 the signal varies (intentionally) with the scan speed, longer dwell time = more photons. Zeiss seems to give out something like counting rates which are then the same for different dwell times. It would be good if the manufacturers give more details. >> >> best wishes >> >> Andreas >> >> >> >> -----Original Message----- >> From: George McNamara <[hidden email]> >> To: [hidden email] >> Sent: Sun, 9 May 2010 14:58 >> Subject: Re: Comparison Zeiss 710 vs Nikon A1 ... full comparison >> >> Hi listserv, >> >> Repeating what I posted in a message yesterday: >> >> I've noticed on both the Leica SP5, Zeiss LSM510 and Zeiss LSM710 that "faster is brighter" for many fluorophores. That is, a 0.4 us dwell time results in a brighter image than a 4 us which is brighter than a 40 us (then repeated 0.4 us, which resulted in the same brightness as the first 0.4 us dwell time - so not photobleaching -- not all fluorophores in the specimen [Desmid slide from Carolina] changed brightness). This implies that <0.4 us might be even brighter - i.e. resonant scanner mode. A couple of possible explanations (not mutually exclusive): >> a) photophysics (possibly caused by fluorophore-O2 photochemistry) - re TRex (PubMed 19337661), papers by Sanden/Spielmann/Widergren et al (PubMed 20375039, 20196585, 19007245, 17385841). >> b) calibration method(s) by Zeiss and/or Leica that try to (but do not always) match output at all settings. >> c) other??? (suggestions/comments welcome!). >> See also >> >> high speed scanning has the potential to increase fluorescence yield and to reduce photobleaching. Borlinghaus RT. Microsc Res Tech. 2006 Sep;69(9):689-92.PMID: 16878313 >> >> Since I can obtain brighter or dimmer fluorescence on the same field of view on the same microscope system by simply changing scan speed, I submit that between microscope comparisons are just anecdotes unless every variable is controlled and reported. Even then there may be variables under the hood that the confocal manufacturer has not mentioned, such as the possibility that the system is "calibrated" to all scan speeds output more or less the same intensity. >> >> >> >> Comments with respect to the post below... >> >> >> At 03:43 PM 4/11/2010, you wrote: >> >>> Hi Pedro, >>> >>> Seems you have short listed A1 and 710. Both the systems are released but sufficiently exposed with A1. I have mentioned my opinion and observation about these two systems earlier that is given below. >>> >>> A1 gives 4X4K image size, whereas 710 gives 6X6K. But do we really need 6KX6K? When we tried taking 6K images, we have experienced photo bleaching in some attempts. >> >> GM: With a 100x1.4+ NA objective lens at the LSM710's 0.6x zoom (and perfect specimen, i.e. refractive index matching and/or imaging right at the coverglass ... and coverglass 170.0 um) >> >> XY resolution = 0.6 * 405 nm (the laser line) / 1.4 = 173 nm (assuming pinhole 1.0 Airy unit, infinitely bright specimen could put sqrt(2) in the denominator). >> Nyquist sampling for a 2D image; 173 nm / 3 = 57 nm. >> >> (I'm not at the LSM710 so don't have the actual image field of view, but this should be close): >> >> For 100x lens and 0.6x zoom, field of view should be about 160 um, so >> >> 160 um / 6000 pixels = 0.026 um = 26 nm. >> >> So, 6kx6k is oversampling by about a factor of two. For a very stable specimen (photostability and vibration isolation), perfect imaging conditions, lots of time (since probably will need Z-series for best deconvolution) and optimized confocal deconvolution algorithm, the 6kx6k setting might be useful. How much more useful than 4kx4k ... please do the experiment, publish and send me the reference. >> >> GM Most confocal microscope software have "optimize" XY resolution buttons - when in doubt, click on it. Incidentally, Paul Goodwin, Applied Precision, told me Biotium CF405 fluorophore is very good for immunofluorescence (context: OMX nanoscope). Might be a good starting point for an experiment such as above. DAPI or Hoechst 33342 or 33358 in a perfect mounting medium could also be a good choice for the above. >> >>> A1 has continuously variable zoom up to 1000X but LSM 710 is limited to 50X. However, we were not given proper training about how to use variable zoom up to 1000X. In my view, the 1000X variable zoom is not that important factor. >>> >>> Fortunately, in 710 there is a master pinhole unlike the earlier 510 that takes time for alignment. However, 710 uses conventional rectangular pinhole but A1 has unique hexagonal pinhole resulting better images. We have checked the same sample in both the systems. >> >> GM: Managing both a 4-pinhole LSM510 and a LSM710, I teach users the same thing on both for multi-color fluorescence (ex. "colocalization"): for the longest wavelength emission channel, select 1 Airy button, then match the pinhole for the other channels. This way the (nominal) optical section thickness is matched. In multitrack mode the pinhole setting can be changed between tracks. >> >> >>> The spectral detector of LSM 710 has 34 channels with simultaneously acquisition unlike the previous model 510 that was with 4x8=32. However, the spectral step size is 3nm (3x34=102nm wavelength resolution) and it is not flexible like A1 that has 2.5nm, 6nm and 32nm (resulting 2.5x32=80nm, 6x32=192nm and 10x32= 320nm wavelength resolution). A1 has surprisingly effective unmixing efficiency even in the close range. >> >> GM: LSM710/ZEN's 32-channel QUASAR detector default is ~10 nm, but is adjustable (5 and 2.5 nm if I recall). When using the flanking PMTs the spectral window for these can be adjusted in 1 nm steps - this is done with two prism/slits - see the LSM710 internals schematic on the Zeiss web site. Going to 1 nm might be useful to play around with the peaks vs off-peaks for Europium or Terbium (which have long fluorescence lifetimes so scan speed would need to be very slow, might be interesting if you have the time to do a spectral scan overnight). >> >>> In LSM710 high wavelength range up to 1100 nm is possible for optimized transmission. I am not sure if the A1 has the same capability. In our (little) experience, 710 gives wonderful sensitivity. Their software Zen 2009 is compatible with Vista/Windows 7. Even anisotropy imaging is possible with LSM 710 (do not know about A1). But you need to purchase this module (extra cost). >>> >>> Nikon software is robust has almost all of the regular modules. However, for Zen we need to pay extra for the add on modules. Some of the regular functions are "optional" with Zen. >>> >>> For simultaneous photo activation and imaging, one need to incorporate Duo system (two heads) into LSM 710 that adds up the cost. However, A1R scan head serves this purpose without any cumbersome modifications and the speed and performance is relatively incomparable. Though there are some annoying "noise" is generated while we use the resonant scanner the speed and performance is still impressive. >>> >>> If you are regularly going for live cell imaging and Ca++ imaging, I feel A1 has so many features and it is not bleaching the dyes or induce unexpected phototoxicity. Though, we found 710 has slightly better sensitivity, we often face bleaching and laser induced toxicity problems for the same set of experiments. >> >> GM: See top of message - scan speed, resolution, laser power at specimen, need to be matched to make valid comparison. >> >> >>> Eager to know the inputs of other users. >>> >>> No commercial interest. >>> >>> Roshma. >>> >>> >>> >>> On Sun, Apr 11, 2010 at 4:34 PM, Pedro J Camello <[hidden email]> wrote: >>> Hi all, >>> >>> has anybody in the list compared Nikon A1 vs Zeiss 710. We´re purchasing a >>> spectral micro with a TIRF module and motorization. Any input will be >>> really wellcome (off list if you prefer) >>> >>> A second question, what is the most close to "live cell" sample to make >>> real tests in confocal? I´m going to travel to test a couple of micros, >>> and to carry or prepapre real living cells is rather complicated for us. >>> We´re especially interested in ion (Ca2+) experiments. >>> >>> Thanks >>> >>> >>> -- >>> Dr Pedro J Camello >>> Dpt Physiology >>> Faculty of Veterinary Sciences >>> University of Extremadura >>> 10071 Caceres >>> Spain >>> Ph: 927257000 Extension 51321/51290 >>> Fax:927257110 >>> >> >> >> >> >> >> >> George McNamara, Ph.D. >> Image Core Manager >> Analytical Imaging Core Facility >> University of Miami, Miller School of Medicine >> Miami, FL 33136 >> [hidden email] >> [hidden email] >> 305-243-8436 office >> http://www.sylvester.org/AICF (Analytical Imaging Core Facility) >> http://www.sylvester.org/AICF/pubspectra.zip (the entire 2000+ spectra >> http://home.earthlink.net/~geomcnamara > > > >Glen MacDonald >Core for Communication Research >Virginia Merrill Bloedel Hearing Research Center >Box 357923 >University of Washington >Seattle, WA 98195-7923 USA >(206) 616-4156 >[hidden email] |
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Andreas Bruckbauer |
Re: Comparison Zeiss 710 vs Nikon A1 ... full comparison requires matching resolution and scan speed
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The "problem" with photon counting mode is that you loose your gain parameter to balance the different channels against each other, maybe the manufacturers should implement autoscaling functions which map the display to the output as it is done for camea based detection or in scanning probe microscopy.
best wishes
Andreas -----Original Message-----
From: Stanislav Vitha <[hidden email]> To: [hidden email] Sent: Tue, 11 May 2010 14:57 Subject: Re: Comparison Zeiss 710 vs Nikon A1 ... full comparison requires matching resolution and scan speed I would expect that in the photon counting (PC) mode all the different |
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