Difference between revisions of "OpenFlexure Scope"

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Our first real build of an OpenFlexure Scope happened quite some time ago (23may19)!
+
Our first real build of an [https://openflexure.org/ OpenFlexure] Scope happened quite some time ago (23may19)!
more to come! <br>
+
 
Following instructions from GitHub
+
However, we plan much more to come, especially for the '''<nowiki/>'Cheek Cell chip' project''', to combine micronuclei and comet assay measurements for a good [[AGiR! for genomic integrity|estimation of DNA damage]] levels!
 +
 
 +
<br>
 +
We basically followed instructions from GitHub.
 
<br>
 
<br>
 
[http://rwb27.github.io/openflexure_microscope/docs/1_actuator_assembly.html Actuator Assembly]
 
[http://rwb27.github.io/openflexure_microscope/docs/1_actuator_assembly.html Actuator Assembly]
Line 7: Line 10:
 
Used Blue LED below, with 150ohm resister to start (maybe not best?) <br>
 
Used Blue LED below, with 150ohm resister to start (maybe not best?) <br>
 
Used longer screws (15mm) to attach the motors. <br>
 
Used longer screws (15mm) to attach the motors. <br>
[[File:Addingmotors.JPG|250px|thumb]]<br>
+
[[File:Addingmotors.JPG|250px|thumb]]<br>The program openflexure ev is put into the fedora system of the 'inner sanctum' space.<br>
The program openflexure ev is put into the fedora system of the 'inner sanctum' space.<br>
 
 
login with pi@openflexture.local  (and pass from our dear old mascot! )<br>
 
login with pi@openflexture.local  (and pass from our dear old mascot! )<br>
The motor board was set up by Masoud and running already by December 2019, [[File:Withmotorboard.jpeg|250px|thumb]]but we are not yet able to take overall panoramas of samples.<br>
+
The motor board was set up by Masoud and running already by December 2019, [[File:Withmotorboard.jpeg|250px|thumb]]but we were not yet able to take overall panoramas of samples.<br>
Figured out exactly how to wire things up, so could do bright field or epifluor.<br>
+
Figured out exactly how to wire things up, so could do bright field or epifluor, and we played with different powers to the blue LED to get more excitation light..<br>
However, to date (dec2019), we have only been able to image nuclei stained with sybr safe, not yet comets...<br>
+
There is a lot of old information in [https://forum.openhardware.science/t/openflexure-microscope-epifluorescence-acquisition/2099/22 this thread of the GOSH Forum].<br>In fact, we played with more power to the blue 'excitation' LED, but it was a filter problem that got resolved before the pandemic that got us able to see fluor nuclei.
 +
 
 +
This is the filter set needed to make this work on sybr-safe stained cells.
 +
 
 +
*1 550nm dichroic mirror, 25x16mm (GFP) Mfr: Comar; Part No. 550 1Y 116
 +
*1 490nm excitation filter, 25x16mm (GFP) Mfr: Comar; Part No. 495 1K 116
 +
*1 500nm emission filter, a ‘long-pass’ Comar 510 IY 116
 +
 
 +
 
 
[[File:Epifluor with OpenFlexure.png|250px|thumb]]
 
[[File:Epifluor with OpenFlexure.png|250px|thumb]]
Much more will come soon, we thought, before we we struck by the SARS-CoV-2 pandemic, including the full configuration so far... <br>
+
 
(thanks to the [www.openflexure.org OpenFlexure] crew at Bath, UK (and elsewhere!), and to all Octanis members, and Masoud for getting the motors to work!!)<br>
+
 
The latest plan would be to try the more sensitive Raspberry Pi camera and the more stable filter insertion version, to see if we can also visualise the comet tails...
+
However, as of dec2019, we were only able to image nuclei stained with sybr safe, not yet actual comet tails...
 +
 
 +
Much more would come soon, we thought, before we we struck by the SARS-CoV-2 pandemic, including the full configuration so far... <br>
 +
(thanks to the [www.openflexure.org OpenFlexure] crew at Bath, UK (and elsewhere!), and to all Octanis members, and Masoud for getting the motors to work!!)
 +
 
 +
<br>
 +
The latest plan, as we are hope we are finally seeing the light at the end of the tunnel out of this pandemic time (March 2022), will be to try the a higher magnification lens that is more sensitive to fluorescence (20x FLUO-TAR), in place of the 10x objective used in our first tests. We also want to use [https://www.raspberrypi.com/products/raspberry-pi-high-quality-camera/ more sensitive Raspberry Pi camera] and the more stable filter insertion epi-fluor version of OpenFlexure, to see if we can also visualise the comet tails...
 +
 
 +
We actually got some support to do it together  **([https://forum.openhardware.science/t/apply-here-for-gosh-s-2022-regional-events-funding-round-2/3511/6&#x20;&#x20;GOSH&#x20;Community&#x20;ForumGOSH&#x20;Community&#x20;Forum asked for GOSH funding, March 2022], and maybe we will even get more people from the GOSH community to participate...)
 +
[[File:OpenFlexure2022.jpg|thumb|brighter than before, perhaps?  comets next!]]
 +
We did, at least, get some epifluorescence of nuclei again! see the montage, below right...
 +
 
 +
=)
 +
 
 +
Then, if all goes well, the 'cheek cell chip' fluidic prototypes would get incorporated in place of the current slide holder, and all our hopes will finally get tested for DNA damage quantitation by both the micronuclei and comets from the same cell samples, and the fun can really begin (chocolate? blueberries?) with the Cheek Cell Chip!
 +
 
 +
You can help!
 +
 
 +
Here is the [[AGiR! for genomic integrity|original Genomic integrity project page]] at Hackuarium, for more info.
 +
 
 +
[[Category:Work In Progress]]

Latest revision as of 15:37, 2 December 2022

Our first real build of an OpenFlexure Scope happened quite some time ago (23may19)!

However, we plan much more to come, especially for the 'Cheek Cell chip' project, to combine micronuclei and comet assay measurements for a good estimation of DNA damage levels!


We basically followed instructions from GitHub.
Actuator Assembly Used WD40 to oil the screws before twisting down.
Used Blue LED below, with 150ohm resister to start (maybe not best?)
Used longer screws (15mm) to attach the motors.

Addingmotors.JPG


The program openflexure ev is put into the fedora system of the 'inner sanctum' space.

login with pi@openflexture.local (and pass from our dear old mascot! )

The motor board was set up by Masoud and running already by December 2019,

Withmotorboard.jpeg

but we were not yet able to take overall panoramas of samples.

Figured out exactly how to wire things up, so could do bright field or epifluor, and we played with different powers to the blue LED to get more excitation light..
There is a lot of old information in this thread of the GOSH Forum.
In fact, we played with more power to the blue 'excitation' LED, but it was a filter problem that got resolved before the pandemic that got us able to see fluor nuclei.

This is the filter set needed to make this work on sybr-safe stained cells.

  • 1 550nm dichroic mirror, 25x16mm (GFP) Mfr: Comar; Part No. 550 1Y 116
  • 1 490nm excitation filter, 25x16mm (GFP) Mfr: Comar; Part No. 495 1K 116
  • 1 500nm emission filter, a ‘long-pass’ Comar 510 IY 116


Epifluor with OpenFlexure.png


However, as of dec2019, we were only able to image nuclei stained with sybr safe, not yet actual comet tails...

Much more would come soon, we thought, before we we struck by the SARS-CoV-2 pandemic, including the full configuration so far...
(thanks to the [www.openflexure.org OpenFlexure] crew at Bath, UK (and elsewhere!), and to all Octanis members, and Masoud for getting the motors to work!!)


The latest plan, as we are hope we are finally seeing the light at the end of the tunnel out of this pandemic time (March 2022), will be to try the a higher magnification lens that is more sensitive to fluorescence (20x FLUO-TAR), in place of the 10x objective used in our first tests. We also want to use more sensitive Raspberry Pi camera and the more stable filter insertion epi-fluor version of OpenFlexure, to see if we can also visualise the comet tails...

We actually got some support to do it together **(asked for GOSH funding, March 2022, and maybe we will even get more people from the GOSH community to participate...)

brighter than before, perhaps? comets next!

We did, at least, get some epifluorescence of nuclei again! see the montage, below right...

=)

Then, if all goes well, the 'cheek cell chip' fluidic prototypes would get incorporated in place of the current slide holder, and all our hopes will finally get tested for DNA damage quantitation by both the micronuclei and comets from the same cell samples, and the fun can really begin (chocolate? blueberries?) with the Cheek Cell Chip!

You can help!

Here is the original Genomic integrity project page at Hackuarium, for more info.