PSF of beads far way from glass surface
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Volodymyr Nechyporuk-Zloy |
PSF of beads far way from glass surface
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To join, leave or search the confocal microscopy listserv, go to: http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy Post images on http://www.imgur.com and include the link in your posting. ***** Dear Colleagues, I would like to acquire PSF of beads far away from glass surface, e.g. 50-100 um, in ibidi chambers. What transparent hard gel or any other media should I use to reduce [eliminate] Brownian motion? Best regards, Volodymyr Dr. Volodymyr Nechyporuk-Zloy (Imaging Facility Manager) My office: 668.30.06D The Kennedy Institute of Rheumatology University of Oxford, Roosevelt Drive, Headington, Oxford, OX3 7FY, United Kingdom Office: (+44) 1865612665 E-mails: [hidden email] [hidden email] Skype: vzloy1work Professional Web: http://focus-on-single-molecule.info Department Web: http://www.kennedy.ox.ac.uk/ |
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Eric Marino |
Re: PSF of beads far way from glass surface
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To join, leave or search the confocal microscopy listserv, go to: http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy Post images on http://www.imgur.com and include the link in your posting. ***** Good Morning Volodymyr, Try suspending them in 1% agarose. Eric Marino [hidden email] On 9/16/16, 7:03 AM, "Confocal Microscopy List on behalf of Volodymyr Nechyporuk-Zloy" <[hidden email] on behalf of [hidden email]> wrote: ***** To join, leave or search the confocal microscopy listserv, go to: http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy Post images on http://www.imgur.com and include the link in your posting. ***** Dear Colleagues, I would like to acquire PSF of beads far away from glass surface, e.g. 50-100 um, in ibidi chambers. What transparent hard gel or any other media should I use to reduce [eliminate] Brownian motion? Best regards, Volodymyr Dr. Volodymyr Nechyporuk-Zloy (Imaging Facility Manager) My office: 668.30.06D The Kennedy Institute of Rheumatology University of Oxford, Roosevelt Drive, Headington, Oxford, OX3 7FY, United Kingdom Office: (+44) 1865612665 E-mails: [hidden email] [hidden email] Skype: vzloy1work Professional Web: http://focus-on-single-molecule.info Department Web: http://www.kennedy.ox.ac.uk/ |
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Carina Monico |
Re: PSF of beads far way from glass surface
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In reply to this post by Volodymyr Nechyporuk-Zloy
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To join, leave or search the confocal microscopy listserv, go to: http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy Post images on http://www.imgur.com and include the link in your posting. ***** Hi Volodymyr, you could try with agarose. You can find a protocol in this paper: www.nature.com/articles/srep16088 Carina ________________________________ ________________________________ Dr. Carina Monico Postdoctoral Research Associate Chemistry Research Laboratory 12 Mansfield Road University of Oxford Oxford OX1 3TA T: 01865 275486 E: [hidden email] ________________________________________ From: Confocal Microscopy List [[hidden email]] on behalf of Volodymyr Nechyporuk-Zloy [[hidden email]] Sent: Friday, September 16, 2016 12:03 PM To: [hidden email] Subject: PSF of beads far way from glass surface ***** To join, leave or search the confocal microscopy listserv, go to: http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy Post images on http://www.imgur.com and include the link in your posting. ***** Dear Colleagues, I would like to acquire PSF of beads far away from glass surface, e.g. 50-100 um, in ibidi chambers. What transparent hard gel or any other media should I use to reduce [eliminate] Brownian motion? Best regards, Volodymyr Dr. Volodymyr Nechyporuk-Zloy (Imaging Facility Manager) My office: 668.30.06D The Kennedy Institute of Rheumatology University of Oxford, Roosevelt Drive, Headington, Oxford, OX3 7FY, United Kingdom Office: (+44) 1865612665 E-mails: [hidden email] [hidden email] Skype: vzloy1work Professional Web: http://focus-on-single-molecule.info Department Web: http://www.kennedy.ox.ac.uk/ |
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In reply to this post by Volodymyr Nechyporuk-Zloy
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Depends on what you want to have in between. Beads can be attached to glass on the top surface (I am not sure if the geometry of your chambers allow that) from 1% polyvinyl acetate in methanol and then you can add water, oil or nothing (Journal ofMicroscopy, Vol. 241, Pt 1 2011, pp. 94–100)
From: Confocal Microscopy List <[hidden email]> on behalf of Volodymyr Nechyporuk-Zloy <[hidden email]>
Sent: Friday, September 16, 2016 7:03 AM To: [hidden email] Subject: PSF of beads far way from glass surface *****
To join, leave or search the confocal microscopy listserv, go to: http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
Post images on http://www.imgur.com and include the link in your posting. ***** Dear Colleagues, I would like to acquire PSF of beads far away from glass surface, e.g. 50-100 um, in ibidi chambers. What transparent hard gel or any other media should I use to reduce [eliminate] Brownian motion? Best regards, Volodymyr Dr. Volodymyr Nechyporuk-Zloy (Imaging Facility Manager) My office: 668.30.06D The Kennedy Institute of Rheumatology University of Oxford, Roosevelt Drive, Headington, Oxford, OX3 7FY, United Kingdom Office: (+44) 1865612665 E-mails: [hidden email] [hidden email] Skype: vzloy1work Professional Web: http://focus-on-single-molecule.info Department Web: http://www.kennedy.ox.ac.uk/ |
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Cvic Innocent |
Re: PSF of beads far way from glass surface
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Hey Volodymyr For a full matrix of beads at various depths I suggest using CyGel. We have a bit over at Micron so if you'd care to testdrive then come over. Cordially, cvic Cassandravictoria Innocent, PhD Micron ABU // Schermelleh Group Dept of Biochemistry Oxford University On Fri, Sep 16, 2016 at 1:13 PM, MODEL, MICHAEL <[hidden email]> wrote: ***** To join, leave or search the confocal microscopy listserv, go to: http://lists.umn.edu/cgi-bin/ |
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Volodymyr Nechyporuk-Zloy |
Re: PSF of beads far way from glass surface
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In reply to this post by Volodymyr Nechyporuk-Zloy
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To join, leave or search the confocal microscopy listserv, go to: http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy Post images on http://www.imgur.com and include the link in your posting. ***** I have tried CyGel. It works to immobilise cells, but for beads it does not work - at least, under room temperature. The beads are moving. |
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Michael Giacomelli |
Re: PSF of beads far way from glass surface
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I've used both PDMS and optical adhesive to immobilize beads. The resulting phantoms are then stable indefinitely. Mike On Fri, Sep 16, 2016 at 9:11 AM, Volodymyr Nechyporuk-Zloy <[hidden email]> wrote: ***** |
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Craig Brideau |
Re: PSF of beads far way from glass surface
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Sylgard completely solidifies and is fairly clear. Not sure what its exact index is but it seems to work well. Craig On Fri, Sep 16, 2016 at 11:05 AM, Michael Giacomelli <[hidden email]> wrote: ***** To join, leave or search the confocal microscopy listserv, go to: http://lists.umn.edu/cgi-bin/ |
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Stanislav Vitha-2 |
Re: PSF of beads far way from glass surface
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In reply to this post by Volodymyr Nechyporuk-Zloy
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To join, leave or search the confocal microscopy listserv, go to: http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy Post images on http://www.imgur.com and include the link in your posting. ***** I normally suspend the beads in 2% agarose, but sometimes the beads still have some limited motion. If you need a higher refractive index than just water, you could suspend the beads in gelatin or acrylamide and when solidified in imaging chamber, infiltrate with whatever mounting media you need. Stan Stanislav Vitha Microscopy and Imaging Center Texas A&M University On Fri, 16 Sep 2016 06:03:15 -0500, Volodymyr Nechyporuk-Zloy <[hidden email]> wrote: > >Dear Colleagues, > >I would like to acquire PSF of beads far away from glass surface, e.g. 50-100 um, in ibidi chambers. What transparent hard gel or any other media should I use to reduce [eliminate] Brownian motion? > > >Best regards, >Volodymyr > > |
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Zdenek Svindrych-2 |
Re: PSF of beads far way from glass surface
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Agarose is quite forgiving, it will solidify in a range of media, including quite high concentrations (70% v/v) of glycerol or TDE. Cooling below room temperature may be needed for better jelling. I haven't tried something like 95% TDE, but once I've left a piece of TDE-agarose dry out on the desk, most of the water evaporated, and I got nice piece of jelly with n=1.51...
Best, zdenek Zdenek Svindrych, Ph.D. W.M. Keck Center for Cellular Imaging (PLSB 003) University of Virginia, Charlottesville, VA http://www.kcci.virginia.edu/ tel: 434-982-4869 Annual FRET Workshop: http://kcci.virginia.edu/workshop-2017 ---------- Původní zpráva ---------- ***** |
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Volodymyr Nechyporuk-Zloy |
Re: PSF of beads far way from glass surface
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In reply to this post by Volodymyr Nechyporuk-Zloy
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To join, leave or search the confocal microscopy listserv, go to: http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy Post images on http://www.imgur.com and include the link in your posting. ***** Dear Michael, What adhesive have you used? Best regards, Volodymyr |
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To join, leave or search the confocal microscopy listserv, go to:
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Hi Volodymyr,
Polyvinyl acetate is a polymer soluble in methanol but not in water. You can mix beads into 1% PVA in methanol, then put a drop of it on a glass slide/coverslip and let it dry. Then the beads remain firmly trapped in a thin film of polymer stuck to the slide and will stay there after you add a liquid (any liquid that will not dissolve PVA).
Best regards
Mike
From: Confocal Microscopy List <[hidden email]> on behalf of Volodymyr Nechyporuk-Zloy <[hidden email]>
Sent: Tuesday, September 20, 2016 6:18 AM To: [hidden email] Subject: Re: PSF of beads far way from glass surface *****
To join, leave or search the confocal microscopy listserv, go to: http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
Post images on http://www.imgur.com and include the link in your posting. ***** Dear Michael, What adhesive have you used? Best regards, Volodymyr |
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