Fluorescent grid (in z) slide

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zhouzc zhouzc
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Fluorescent grid (in z) slide

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Hi all,

I'm trying to measure how grin lenses distort and magnify the image in the z-axis. I know there are 2D fluorescent grids that are evenly spaced (eg. https://www.psfcheck.com/about-us) for lateral measurements, but I was wondering if there any similar slides/samples but for axial measurements.

Thanks!
Charles
Armstrong, Brian Armstrong, Brian
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Re: Fluorescent grid (in z) slide

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Hi Charles, I do not, however I think you may want to image beads. There are many commercially available beads such as Tetraspec beads.

Cheers,

Brian Armstrong PhD
Director, Light Microscopy Core
Associate Research Professor
Department of Shared Resources
Department of Diabetes and Metabolic Diseases
Beckman Research Institute, City of Hope

-----Original Message-----
From: Confocal Microscopy List <[hidden email]> On Behalf Of zhouzc
Sent: Thursday, March 11, 2021 8:39 AM
To: [hidden email]
Subject: Fluorescent grid (in z) slide

[Attention: This email came from an external source. Do not open attachments or click on links from unknown senders or unexpected emails.]

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Post images on https://urldefense.com/v3/__http://www.imgur.com__;!!Fou38LsQmgU!_9dErtqxNLTVw93MEde7LwaTBKeKZH-7ly1JqHverfzzNtYBDp671KL353i-P98$  and include the link in your posting.
*****

Hi all,

I'm trying to measure how grin lenses distort and magnify the image in the z-axis. I know there are 2D fluorescent grids that are evenly spaced (eg. https://urldefense.com/v3/__https://www.psfcheck.com/about-us__;!!Fou38LsQmgU!_9dErtqxNLTVw93MEde7LwaTBKeKZH-7ly1JqHverfzzNtYBDp671KL3NClVGqY$ ) for lateral measurements, but I was wondering if there any similar slides/samples but for axial measurements.

Thanks!
Charles

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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. (LCP301)
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Pieter de Tombe Pieter de Tombe
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Re: Fluorescent grid (in z) slide

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Try Argolight, expensive though but it works well.  
Pieter

Sent from my iPhone

> On Mar 11, 2021, at 6:11 PM, Brian Armstrong <[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.
> *****
>
> Hi Charles, I do not, however I think you may want to image beads. There are many commercially available beads such as Tetraspec beads.
>
> Cheers,
>
> Brian Armstrong PhD
> Director, Light Microscopy Core
> Associate Research Professor
> Department of Shared Resources
> Department of Diabetes and Metabolic Diseases
> Beckman Research Institute, City of Hope
>
> -----Original Message-----
> From: Confocal Microscopy List <[hidden email]> On Behalf Of zhouzc
> Sent: Thursday, March 11, 2021 8:39 AM
> To: [hidden email]
> Subject: Fluorescent grid (in z) slide
>
> [Attention: This email came from an external source. Do not open attachments or click on links from unknown senders or unexpected emails.]
>
> ----------------------------------------------------------------------
> *****
> To join, leave or search the confocal microscopy listserv, go to:
> https://urldefense.com/v3/__http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy__;!!Fou38LsQmgU!_9dErtqxNLTVw93MEde7LwaTBKeKZH-7ly1JqHverfzzNtYBDp671KL3jSBU1l8$ 
> Post images on https://urldefense.com/v3/__http://www.imgur.com__;!!Fou38LsQmgU!_9dErtqxNLTVw93MEde7LwaTBKeKZH-7ly1JqHverfzzNtYBDp671KL353i-P98$  and include the link in your posting.
> *****
>
> Hi all,
>
> I'm trying to measure how grin lenses distort and magnify the image in the z-axis. I know there are 2D fluorescent grids that are evenly spaced (eg. https://urldefense.com/v3/__https://www.psfcheck.com/about-us__;!!Fou38LsQmgU!_9dErtqxNLTVw93MEde7LwaTBKeKZH-7ly1JqHverfzzNtYBDp671KL3NClVGqY$ ) for lateral measurements, but I was wondering if there any similar slides/samples but for axial measurements.
>
> Thanks!
> Charles
>
> ----------------------------------------------------------------------
> ------------------------------------------------------------
> -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. (LCP301)
> ------------------------------------------------------------
Sathya Srinivasan Sathya Srinivasan
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Re: Fluorescent grid (in z) slide

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*****
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*****

Hi Charles,
We have an electronic microcrocater which measures the Z-axis in our
stereology system. The Z-axis measurement is very important for
stereological studies. There are a few published methods for Z axis
measurements:

-
https://www.agarscientific.com/media/import/03_Calibration_Standards_pgs_39-84_date_17_06_10_web.pdf
(have different types of standards for X, Y and Z calibration including
beads of different sizes)
- Disector Z-axis mechanical method for stereology (
https://www.scielo.br/pdf/aabc/v82n2/28.pdf)
- A three-dimensional calibration device for the confocal microscope (
https://pdfs.semanticscholar.org/11ca/316460cfa41672f817388d0dbfa3d43f7cd8.pdf
)
- Procedure for Calibrating the Z-axis of a Confocal Microscope-
Application for the Evaluation of Structured Surfaces (
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6387104/pdf/sensors-19-00527.pdf
)
- Calibration of the automated Z-axis of a microscope using focus functions
(
https://d1rkab7tlqy5f1.cloudfront.net/TNW/Over%20faculteit/Decaan/Publications/1997/JoM97FBLVTY.pdf
)
- Industrial Calibration Procedure for Confocal Microscopes (
https://www.mdpi.com/1996-1944/12/24/4137/pdf)

You can use your imagination with some fluorescence coating on surfaces at
different depths for your measurements. Not sure if all the links provided
above will work.

Good luck.

Sathya Srinivasan
ONPRC

On Thu, Mar 11, 2021 at 8:55 AM zhouzc <[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.
> *****
>
> Hi all,
>
> I'm trying to measure how grin lenses distort and magnify the image in the
> z-axis. I know there are 2D fluorescent grids that are evenly spaced (eg.
> https://www.psfcheck.com/about-us) for lateral measurements, but I was
> wondering if there any similar slides/samples but for axial measurements.
>
> Thanks!
> Charles
>
Steffen Dietzel Steffen Dietzel
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Re: Fluorescent grid (in z) slide

In reply to this post by zhouzc
*****
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.
*****

the Argolight slide has a sphere made of 3 rings: one in xy, one in xz
and one in yz. Also, there are several z-stair patterns on it, with
different z-step sizes. It sounds like you would prefer a cube with a
3D-dot pattern (which might be a good idea for future versions) but
maybe one of those patterns would do. argolight.com

It is not cheap, but it gives you a lot of options for quality control.
If you should have money to burn, you might get a custom pattern. No
commercial interest on my side.

Out of curiosity. should you not be able to see 3D distortion if you
focus through a 2D pattern? Not that I would know much about grin
lenses. At the end of the day, every available detector just takes a 2D
image at max, and assembles 3D in the computer.

Steffen


Am 11.03.2021 um 17:39 schrieb zhouzc:

> *****
> 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 all,
>
> I'm trying to measure how grin lenses distort and magnify the image in
> the z-axis. I know there are 2D fluorescent grids that are evenly
> spaced (eg. https://www.psfcheck.com/about-us) for lateral
> measurements, but I was wondering if there any similar slides/samples
> but for axial measurements.
>
> Thanks!
> Charles

--
------------------------------------------------------------
Steffen Dietzel, PD Dr. rer. nat
Ludwig-Maximilians-Universität München
Biomedical Center (BMC)
Head of the Core Facility Bioimaging

Großhaderner Straße 9
D-82152 Planegg-Martinsried
Germany

http://www.bioimaging.bmc.med.uni-muenchen.de
Zdenek Svindrych-2 Zdenek Svindrych-2
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Re: Fluorescent grid (in z) slide

*****
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http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
Post images on http://www.imgur.com and include the link in your posting.
*****

Hi Steffen,
one use case where z-calibration is non-trivial is when the GRIN is fixed
to the sample and you're focusing with a movable objective lens on the
other side of the GRIN (or more exotic arrangements).
Also, GRIN lenses (at least those I worked with) cannot be made
telecentric, so the PSF is not constant over the field of view...(pain)

Charles:
For the z-calibration I was thinking about some layers of tape (or thin
coverslip) with some beads in between, but the z-spacing will be rather
coarse (50 - 100 um at least) and not very uniform.

Besty, zdenek

On Fri, Mar 12, 2021 at 3:36 AM Steffen Dietzel <[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.
> *****
>
> the Argolight slide has a sphere made of 3 rings: one in xy, one in xz
> and one in yz. Also, there are several z-stair patterns on it, with
> different z-step sizes. It sounds like you would prefer a cube with a
> 3D-dot pattern (which might be a good idea for future versions) but
> maybe one of those patterns would do. argolight.com
>
> It is not cheap, but it gives you a lot of options for quality control.
> If you should have money to burn, you might get a custom pattern. No
> commercial interest on my side.
>
> Out of curiosity. should you not be able to see 3D distortion if you
> focus through a 2D pattern? Not that I would know much about grin
> lenses. At the end of the day, every available detector just takes a 2D
> image at max, and assembles 3D in the computer.
>
> Steffen
>
>
> Am 11.03.2021 um 17:39 schrieb zhouzc:
> > *****
> > 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 all,
> >
> > I'm trying to measure how grin lenses distort and magnify the image in
> > the z-axis. I know there are 2D fluorescent grids that are evenly
> > spaced (eg. https://www.psfcheck.com/about-us) for lateral
> > measurements, but I was wondering if there any similar slides/samples
> > but for axial measurements.
> >
> > Thanks!
> > Charles
>
> --
> ------------------------------------------------------------
> Steffen Dietzel, PD Dr. rer. nat
> Ludwig-Maximilians-Universität München
> Biomedical Center (BMC)
> Head of the Core Facility Bioimaging
>
> Großhaderner Straße 9
> D-82152 Planegg-Martinsried
> Germany
>
> http://www.bioimaging.bmc.med.uni-muenchen.de
>


--
--
Zdenek Svindrych, Ph.D.
Research Scientist - Microscopy Imaging Specialist
Department of Biochemistry and Cell Biology
Geisel School of Medicine at Dartmouth
zhouzc zhouzc
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Re: Fluorescent grid (in z) slide

*****
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 all,

Great ideas - I'm looking into the argolight slides and I think your solution Zdenek will be a nice interim method.

Thanks all!
Charles

-----Original Message-----
From: Confocal Microscopy List <[hidden email]> On Behalf Of Zdenek Svindrych
Sent: Friday, March 12, 2021 8:42 AM
To: [hidden email]
Subject: Re: Fluorescent grid (in z) slide

*****
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 Steffen,
one use case where z-calibration is non-trivial is when the GRIN is fixed to the sample and you're focusing with a movable objective lens on the other side of the GRIN (or more exotic arrangements).
Also, GRIN lenses (at least those I worked with) cannot be made telecentric, so the PSF is not constant over the field of view...(pain)

Charles:
For the z-calibration I was thinking about some layers of tape (or thin
coverslip) with some beads in between, but the z-spacing will be rather coarse (50 - 100 um at least) and not very uniform.

Besty, zdenek

On Fri, Mar 12, 2021 at 3:36 AM Steffen Dietzel <[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.
> *****
>
> the Argolight slide has a sphere made of 3 rings: one in xy, one in xz
> and one in yz. Also, there are several z-stair patterns on it, with
> different z-step sizes. It sounds like you would prefer a cube with a
> 3D-dot pattern (which might be a good idea for future versions) but
> maybe one of those patterns would do. argolight.com
>
> It is not cheap, but it gives you a lot of options for quality control.
> If you should have money to burn, you might get a custom pattern. No
> commercial interest on my side.
>
> Out of curiosity. should you not be able to see 3D distortion if you
> focus through a 2D pattern? Not that I would know much about grin
> lenses. At the end of the day, every available detector just takes a
> 2D image at max, and assembles 3D in the computer.
>
> Steffen
>
>
> Am 11.03.2021 um 17:39 schrieb zhouzc:
> > *****
> > 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 all,
> >
> > I'm trying to measure how grin lenses distort and magnify the image
> > in the z-axis. I know there are 2D fluorescent grids that are evenly
> > spaced (eg. https://www.psfcheck.com/about-us) for lateral
> > measurements, but I was wondering if there any similar
> > slides/samples but for axial measurements.
> >
> > Thanks!
> > Charles
>
> --
> ------------------------------------------------------------
> Steffen Dietzel, PD Dr. rer. nat
> Ludwig-Maximilians-Universität München Biomedical Center (BMC) Head of
> the Core Facility Bioimaging
>
> Großhaderner Straße 9
> D-82152 Planegg-Martinsried
> Germany
>
> http://www.bioimaging.bmc.med.uni-muenchen.de
>


--
--
Zdenek Svindrych, Ph.D.
Research Scientist - Microscopy Imaging Specialist Department of Biochemistry and Cell Biology Geisel School of Medicine at Dartmouth
Alex Corbett-2 Alex Corbett-2
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Re: Fluorescent grid (in z) slide

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*****
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*****

Dear Charles,

It is worth mentioning that the standard PSFcheck slide comes with three-dimensional arrays of features (both large and small). The array size is 10 x 10 x 3 features (XYZ) on a 10 um pitch, which should allow you to calibrate in Z as well as XY.

Best regards, Alex Corbett.