WG 1 – Illumination power

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WG 1 – Objectives

Comparison of fluorescence intensities between images requires measurements of the illumination power and stability of the excitation light source. WG1 aims at establishing a recommended protocol for measuring the stability of a light source during both short- and long-term image acquisition sessions using calibrated external power sensors. This initial aim will be extended later to measure the absolute flux of light through the illumination path and irradiance of the sample. The initial protocol will be designed around lasers on confocal microscopy platforms (raster scanning and spinning disks). It will be later modified towards other microscopy techniques (widefield, TIRF, light-sheet, super-resolution).

WG 1 – Co-Chairs

WG 1 - Latest News

WG 1 - Illumination Power published its third version of the protocol "Illumination Power, Stability and Linearity Measurements for Confocal, Widefield and Multiphoton Microscopes". The protocol is featured on protocols.io

We are delighted to warmly welcome our first member from Thailand, Dr Tassanee Lerksuthirat from the Research Center Ramathibodi Hospital just joined Working Group 1, 7, 8, 10, 11 and 12 of QUAREP-LiMi.

WG 1 - Illumination Power published its second version of the protocol "Illumination Power, Stability and Linearity Measurements for Confocal and Widefield Microscopes". The protocol is featured on protocols.io

 

Last meeting: 2nd April 2025, 1:01 pm

Release of protocol “Illumination power and illumination stability measurements for contocal, widefield and multiphoton microscopes“. 

Beside step-by-step instructions, the protocol provides:

  1. Automation
    • Aims: The QUAREP-LiMi power and stability assessment protocol requires long and repetitive measurements for each light source as other parameters are tested. The aim of this subgroup is to provide feasible means for carrying out these processes, taking advantage of the automation capabilities of modern microscopes.
    • Co-leads: Mišo Mitkovski, Nasser Darwish
    • On-going tasks:
    • Cloud storage location (can be accessed as a member) here
  2. Environment
  3. Glossary
  4. Power Meter

40th Meeting, 2nd of April 2025

Meeting date

02/04/2025 @ 1:01 PM (CET/CEST)

Meeting summary

Thank you to all who attended our 40th meeting this week. We had a total of 14 participants.
  1. We reviewed the slides for the updates of WG1 at the annual meeting and these were approved by the group. You can find these slides here.
  2. Julio presented a prototype of the template that could be used to record the data and metadata related to our protocol in Omero metrics. Our next step will be to test it.
  3. We briefly reviewed our goals and table of contents for the Irradiance manuscript (see below). We need to determine a reasonable timeline for the completion of our first draft. Please use this spreadsheet to indicate your intention to contribute to specific sections. Also, indicate your preferred timeline.
    1. The initial goal and scope have been defined as a guideline and review of what is known and what has been published on the different ways to measure irradiance. The target audience will be biologists.
    2. Table of Content
Introduction Topic 1: What is irradiance? Definitions Topic 2: Why should we care about irradiance? Body Topic 3: How to make the measurement Topic 4: When is this measurement needed? (include examples) Topic 5: Pitfall and issue when irradiance measurements is not performed Discussion Topic 6 (irradiance and phototoxicity): – Topic 7: Important considerations (other than phototoxicity) Topic 8: Recommendations by QUAREP-LiMi.   Conclusion: take-home message   Potential Roles: Writers Editors Reviewers Data/Figure Provider   You can find the complete minutes and recording of the meeting here.   Our next meeting will be on May 7th.

Meeting documentation

39th meeting, 5th of March 2025

Meeting date

05/03/2025 @ 5:00 PM (CET/CEST)

Meeting summary

Thank you to all who attended our meeting last Wednesday. We had a total of 10 participants.   At the SC meeting earlier this week, the code of conduct and WG11 checklist for method reproducibility were finalized and will be shared at the annual meeting. WG1 planned a short update for the annual meeting, focusing on the third version of their protocol and future directions and including an introduction to the automation tools. Julio Mateos Langerak joined our meeting to discuss the feasibility of using Omero metrics to store power measurement data in a centralized database (see the meeting recording and YouTube link below). A combined workshop with WG2 on automation was proposed for the ELMI meeting, with Nasser Darwish and others preparing presentations. Finally, Nathalie will contact people who have signed up for the Irradiance guidance manuscript and set up a way for this subgroup to contribute asynchronously.   You can find the complete minutes and recording of the meeting here.   Our next meeting will be on April 2nd.

Meeting documentation

38th meeting, 5th of February 2025

Meeting date

05/02/2025 @ 3:29 PM (CET/CEST)

Meeting summary

Meeting documentation

37th meeting, 4th of December 2024

Meeting summary

Thank you to all who attended our meeting yesterday. We had a total of 13 participants.   At the SC meeting earlier this week, members voted in favor of the publication of  V3 of our protocol. We will make it public and get a DOI before the holiday break. We discussed and resolved the remaining comments left in protocols.io. We have two remaining items before making the protocol public: 1) Tinging will provide a video tutorial with a multi-photon Leica system and Laurent will add final comments to the protocol as per our discussion.   We also received a written endorsement from WG13 as well as WG7. These can be found here. Judith and Julio presented their proposal on behalf of WG7.  Briefly,  we will work in collaboration with WG7 to create a CSV template with all required fields of metadata and illumination power calibration data needed to be recorded and shared when performing this protocol. This template will be added to a future version of the protocol. We also hope to make it compatible with the automation scripts and power meter software used to run this protocol. This work will begin in February 2025.   You can find the minutes and recording of the meeting here.   Since the first Wednesday of January is on the 1stour next meeting will be on February 5th.

36th meeting, 6th of November 2024

Meeting date

06/11/2024 @ 3:32 PM (CET/CEST)

Meeting summary

Thank you to all who attended our meeting today. We had a total of 11 participants and one new member (Sam Stats).   David Grunwald asked us to review a manuscript from his lab (see previous agenda, containing many references to Quarep-LiMi protocols. He wanted us to ensure nothing contradicted or was not aligned with the QUAREP WG-1 protocol. We reviewed it together and discussed it with David. (please add your comments here) No major issues were found, and overall, everyone welcomed this new tool and can’t wait to be able to test it. We discussed the latest comments received on the final V3 protocol. All comments have been addressed and resolved. Other WG have until mid-November to provide their endorsement or edits. Our next step is to request permission to publish from the SC. We discuss the need to review the skeleton template for the irradiance manuscript. This should help us decide on the content. The next step will be to allocate each section to members of the group. We will set a timeline early next year for this manuscript. You can find the complete minutes of the meeting here.

35th meeting, 2nd of October 2024

Meeting date

02/10/2024 @ 5:00 PM (CET/CEST)

Meeting summary

We had a total of 10 participants. We have finalized Version 3 of our protocol on 2P extension. We have resolved all comments and added a new disclaimer. Our next step is to align with relevant WG and ask them to review and endorse our protocol. Nathalie will prepare a short presentation and will send instructions to the relevant co-chairs. We should complete this part of the review process by the end of October. We have a draft template to structure the manuscript on Irradiance measurements. Please review it before our next meeting. Our next meeting will be on, Wednesday, November 6th, same time, same Zoom link.

Meeting documentation

34th meeting, 4th of September 2024

Meeting date

04/09/2024 @ 5:00 PM (CET/CEST)

Meeting summary

We had a total of 14 participants. Luis is now co-chairing WG1 with Nathalie. We are finalizing Version 3 of our protocol on 2P extension. Laurent has been spearheading this effort. In the second part of the meeting, we discussed and proposed to write a manuscript to educate biologists on irradiance measurements. We will start drafting a skeleton and template for the manuscript at our next meeting. Our next meeting will be on, Wednesday, October 2nd, same time, same Zoom link. The meeting calendar invite series (every first Wednesday of the month) has been sent to everyone.   You can find the complete minutes and the recording of the meeting here. Quick announcement: Laurent shared with us a new peer-reviewed journal that could be a good place for many QUAREP protocols and communications. Methods of Microscopy magazine registration, it is free. The link to the journal can be found here. The SC committee invites you to review the Update of Best Practices document (https://nextcloud.quarep.org/index.php/f/469900) and provide feedback and suggestions. There will be a panel discussion on image data sharing at ASCB 2024 in San Diego. We believe this session will be of great interest and relevant to all industry partners. If you are attending please let Nathalie know and contact her if you want more information.

Meeting documentation

33rd meeting, 3rd of July 2024

Meeting date

03/07/2024 @ 5:00 PM (CET/CEST)

Meeting summary

A small group (9 of us) met to discuss some logistics. We will not meet in August and observe the summer break vacations. We will reconvene in September to complete V3 of our illumination stability protocol and will begin meeting to develop a protocol and guidelines for the measurement of irradiance. Luis Acevedo has volunteered to be a co-chair of WG1 with Nathalie. TingTing has volunteer to represent WG1 at the OB. We still need someone to volunteer to represent WG1 at the ERB. Nathalie will represent WG1 at the SC. The minutes and recoding of the meeting can be found here.

Meeting documentation

32nd Meeting, 3rd of April 2024

Meeting date

03/04/2024 @ 5:00 PM (CET/CEST)

Meeting summary

We had a short round of presentation for new comers. We decided to pause just this time our work on our protocol and 2-photon microscopy modality to focus on irradiance, and had the pleasure to listen to two very nice presentations by Ronian Siew and then Alienor Lahlou. For those of you who missed it, the recordings can be found here.

Meeting documentation

31st meeting, 5th of March 2024

Meeting date

05/03/2024 @ 5:00 PM (CET/CEST)

Meeting summary

We summarized the By-Laws’ presentation since everyone had seen it at least once. The by-laws and its accompanying documents (Code of Conduct – CoC, Terms of Reference – ToR, and Best Practices – BP) aim to ensure QUAREP-LiMi provides a diverse, equitable, inclusive and transparent environment for its community to gather, address and efficiently make decision to critical challenges. You can find previous recordings of the By-Laws presentation from WG2 here, as well as the slides here. In the folder named Decision Making Discussion https://nextcloud.quarep.org/index.php/f/445127 you will find the other relevant documents related to the by-laws: •BP_BestPractices for QUAREP-LiMi Operations.docx •CoC_WorkGroups.docx •Bylaws Draft_04_a.docx Here are our action items: 1.      WG1 needs to appoint one representative for the SC, ERB, and OB (two-year terms). 2.      WG1 needs to elect its co-chairs for the next two-year term. Nathalie and Laurent are happy to continue on for another two year. We would love to have a third co-chair. 3.      We need candidates! – Let us know if you are interested in being our WG1 representative for SC (ideally one of the co-chairs), ERB, or OB, or if you want to become a WG1 co-chair before March 12th. We also briefly brainstormed on known ways to measure irradiance and decided that a path forward would be to review a proposed definition (by WG13) and present a manuscript (journal club style) at our next meeting. WG1 will meet again on the 3rd of April and will then continue expanding the Illumination power stability protocol to multi-photons.

Meeting documentation

Memberlist WG 1