Argolight Argo-HM slides

[Alexia Loynton-Ferrand]

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Dear all,

We recently purchased an Argo-HM slide, to more routinely assess our fluorescent systems.
I am trying to put together some documentation on which patterns to use for which task, and I would like to ask for your feedback.

There seems to be many possibilities, did any of you already prepare some documentation?
How are you using this slide, which pattern did you find the most useful?
For the intensity-sensitive patterns (B, C and D), is it still easy to measure?
Are you using the associated software?

Any feedback welcome :)
Best regards,
Alexia

>>Please note my NEW PHONE NUMBER: [phone] (or alternatively [phone]) <<
------------------------------------
Dr Alexia Loynton-Ferrand | Research Associate in Advanced Light Microscopy | Imaging Core Facility  | Biozentrum, University of Basel | Klingelbergstrasse 50/70 | CH-4056 Basel
Phone: [phone] | [hidden email]<mailto:[hidden email]> | www.biozentrum.unibas.ch<http://www.biozentrum.unibas.ch/> | www.microscopynetwork.unibas.ch<http://www.microscopynetwork.unibas.ch/>

[0000001ed7f52e4a-dmarc-request]

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Dear Alexia and all,

I only briefly tested the Argolight slide and think it would be good to draw up a more precise protocol. I found the resolution pattern very useful as it has lines with different separation and the software then computes a graph contrast vs separation, so one can estimate resolution form a fit of several data points. The software calculates the separation from the measurement and then does the plot of contrast vs. this measured separation. This works quite well for a larger contrast value, like 26% for the Rayleigh criterion, as the lines are well resolved, but for low values e.g. 5% the separation calculated by the software (or the value it obtains in the case of no separation at all) deviates a lot from the specified resolution as the measurement is unreliable. I would prefer a plot against specified resolution, even when the error bars for the specified values are quite large avoiding the unreliable measurements. Furthermore a proper fit with multiple Gaussians might give a better estimate than the peak value. But for relative measurements between instruments and over time at 26% contrast the software works quite well.  

I took z-stacks to get the best focus, but the software seems to work on single images. I think one has to be careful to take images with sufficient SNR, especially on older confocals with less sensitive detectors, otherwise the measured resolution is degraded by low SNR. And of course one has to take the images using Nyquist sampling. It is quite fun to compare raw data with deconvolved images.

The z-ladders are quite good to show the fish-tank effect with air or water lenses, for oil lenses it should be possible to check the z-calibration of the microscope. I have not tested the intensity patterns, but as I understand they are also composed of lines? Not really sure what to do with all the other patterns though. Now there are new slides with fewer patterns which are more affordable then the original one (no commercial interest)!

best wishes

Andreas

 
 
-----Original Message-----
From: Alexia Loynton-Ferrand <[hidden email]>
To: CONFOCALMICROSCOPY <[hidden email]>
Sent: Thu, Jan 25, 2018 3:10 pm
Subject: Argolight Argo-HM slides

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To join, leave or search the confocal microscopy listserv, go to:
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Dear all,

We recently purchased an Argo-HM slide, to more routinely assess our fluorescent systems.
I am trying to put together some documentation on which patterns to use for which task, and I would like to ask for your feedback.

There seems to be many possibilities, did any of you already prepare some documentation?
How are you using this slide, which pattern did you find the most useful?
For the intensity-sensitive patterns (B, C and D), is it still easy to measure?
Are you using the associated software?

Any feedback welcome :)
Best regards,
Alexia

>>Please note my NEW PHONE NUMBER: [phone] (or alternatively [phone]) <<
------------------------------------
Dr Alexia Loynton-Ferrand | Research Associate in Advanced Light Microscopy | Imaging Core Facility  | Biozentrum, University of Basel | Klingelbergstrasse 50/70 | CH-4056 Basel
Phone: [phone] | [hidden email]<mailto:[hidden email]> | www.biozentrum.unibas.ch<http://www.biozentrum.unibas.ch/> | www.microscopynetwork.unibas.ch<http://www.microscopynetwork.unibas.ch/>

[Tim O'Brien Sr.]

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Dear Listserve-

We are excited to announce our 10th annual workshop on the application of, and assessment of the responses to, forces on living cells this May 15-18th, 2018.  This is the third year that our course will incorporate the use of FRET-based biosensors and light modulated signaling molecules from the laboratory of Klaus Hahn, and the application of our vertical light sheet and pathway rotated imaging for sideways microscopy (PRISM) techniques. As usual, there will be morning lectures and afternoon, hands-on labs.  Highlights this year include guest speaker Teng-Leong Chew from HHMI Janelia Research Center, who will present two morning sessions on image analysis using the ImageJ/FIJI toolbox, and the Keynote talk by Wes Legant, recently of the Betzig lab at Janelia, but now a (proud) Tar Heel.  The blurb below describes the course in more detail, and how to apply.

(Note: please plan to stay through the Friday noon session, as the Image Processing presentations really get going on the second day!)

Please feel free to pass this along to anyone that might be interested!

Thanks,
Tim O'Brien
UNC Chapel Hill
-------------------------------------------------------------------------------------------
The Tenth annual Carolina Workshop on Biosensors and Force Measurements in Living Cells is scheduled for May 15th – 18th, 2018.
Cells respond to external biochemical and mechanical cues to generate myriad responses including force generation, motility, gene expression and differentiation. The connection between external cue and internal response are the biochemical pathways that control the cell. Understanding these responses is critical to gaining insight into diverse biomedical phenomena such as cancer, immunity and tissue differentiation and morphogenesis. Our workshop will offer, through morning lectures and afternoon laboratory sessions, ways to assess the biochemistry and mechanobiology of cells, and the theoretical basis for interpreting physical properties, force sensing and force generation.  We will cover the use of FRET-based biosensors related to cell mechanics and responses, and the use of light-modulated signaling molecules. We will present the optics and plans for assembling your own versatile light control microscopy system that includes vertical light sheet microscopy, spinning total internal reflection fluorescence (TIRF) and photoactivation capability.  This year’s workshop will continue to include two morning sessions on image analysis using the FIJI toolbox, taught by guest speaker Teng-Leong Chew from HHMI Janelia Research Center. Experimental stations will include atomic force microscopy combined with light sheet microscopy, 3D force microscopy (magnetic tweezers), FRET microscopy using fluorescent biosensors and the production and use of traction force substrates with soft silicon gels that allow TIRF imaging.  This year our registration fee in only $300, and this includes all supplies, light breakfasts, and a conference dinner/poster session with Dr. Wes Legant as our Keynote Speaker.  The workshop is hosted by CISMM, our NIH/NIBIB resource (Center for Computer Integrated Systems for Microscopy and Manipulation).   To Register and for more information:  http://cismm.web.unc.edu/resources/events//<http://cismm.web.unc.edu/resources/events/>

A flyer that can be printed and put up near your lab is available at:
https://drive.google.com/open?id=1X3rm1wg63rfuk3QMiLV1HOG8Ss0tQCCI

[Tim O'Brien Sr.]

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Dear Listserve-


We wanted to say that we have had a good response to our announcement of this year's Carolina Workshop on Biosensors and Force Measurements in Living Cells thus far, but the good news is that there are still many spaces left.  It promises to be great weather, and awesome science, as usual.  Please tell a friend. The workshop is hosted by CISMM, our NIH/NIBIB resource (Center for Computer Integrated Systems for Microscopy and Manipulation).   To Register and for more information:  http://cismm.web.unc.edu/resources/events//<http://cismm.web.unc.edu/resources/events/>


A flyer that can be printed and put up near your lab is available at:

https://drive.google.com/file/d/1X3rm1wg63rfuk3QMiLV1HOG8Ss0tQCCI/view


Thanks!


Tim O'Brien


I reprint the announcement below:


The Tenth annual Carolina Workshop on Biosensors and Force Measurements in Living Cells is scheduled for May 15th – 18th, 2018.
Cells respond to external biochemical and mechanical cues to generate myriad responses including force generation, motility, gene expression and differentiation. The connection between external cue and internal response are the biochemical pathways that control the cell. Understanding these responses is critical to gaining insight into diverse biomedical phenomena such as cancer, immunity and tissue differentiation and morphogenesis. Our workshop will offer, through morning lectures and afternoon laboratory sessions, ways to assess the biochemistry and mechanobiology of cells, and the theoretical basis for interpreting physical properties, force sensing and force generation.  We will cover the use of FRET-based biosensors related to cell mechanics and responses, and the use of light-modulated signaling molecules. We will present the optics and plans for assembling your own versatile light control microscopy system that includes vertical light sheet microscopy, spinning total internal reflection fluorescence (TIRF) and photoactivation capability.  This year’s workshop will continue to include two morning sessions on image analysis using the FIJI toolbox, taught by guest speaker Teng-Leong Chew from HHMI Janelia Research Center. Experimental stations will include atomic force microscopy combined with light sheet microscopy, 3D force microscopy (magnetic tweezers), FRET microscopy using fluorescent biosensors and the production and use of traction force substrates with soft silicon gels that allow TIRF imaging.  This year our registration fee in only $300, and this includes all supplies, light breakfasts, and a conference dinner/poster session with Dr. Wes Legant as our Keynote Speaker.  The workshop is hosted by CISMM, our NIH/NIBIB resource (Center for Computer Integrated Systems for Microscopy and Manipulation).   To Register and for more information:  http://cismm.web.unc.edu/resources/events//<http://cismm.web.unc.edu/resources/events/>