The IBEX Imaging Knowledge-Base: A Community Resource Enabling Adoption and Development of Immunofluoresence Imaging Methods

Reviewer #1 (Public review):

IBEX Knowledge Database

Here, Anidi and colleagues present the IBEX knowledge base. A community tool developed to centralize knowledge and help its adoption by more users. The authors have done a fantastic job, and there is careful consideration of the many aspects of data management and FAIR principles. The manuscript needs no further work, as it is very well written and has detailed descriptions for data contribution as well as describing the KB itself. Overall, it is a great initiative, especially the aim to inform about negative data and non-recommended reagents, which will positively affect the user community and scientific reproducibility.

As such amount of work has been put into developing this community tool, it would be worth thinking about how it could serve other multiplex-immunofluorescence methods (such as immunoSABER, 4i, etc). Adding an extra tab where the particular method that uses those reagents is mentioned. This would also help as IBEX itself and related methods evolve in the future.

It has a rather minimal description of the software. In particular, there is software that has not been developed for IBEX specifically but that could be used for IBEX datasets (ASHLAR, WSIReg, VALIS, WARPY, and QuPath, etc). It would be nice if there was mention of those.

There is a concern about how the negative data information will be added, as no publication or peer-review process can back it up. Perhaps the particular conditions of the experiment should be very well described to allow future users to assess the validity. The proposed scheme where a reagent can be validated or recommended against by up to 4 different labs should be good. It may be good to make sure that researchers who validate belong to different labs and are not only different ORCID that belong to the same group. Similar to making a case of recommendations against a reagent.

It is very interesting to keep track of the protocol versions used. Perhaps users should be able to validate independent versions and it will be important to know how information is kept.

The final point I would make is that the need to form a GitHub repository may deter some people from submitting data. For sporadic contributions, authors could think that users could either reach out to main developers and/or provide a submission form that can help less experienced users of command-line and GitHub programming, but still promote the contribution from the community.

I am keen to see how the KB evolves and how it helps disseminate the use of this and other great techniques.

Reviewer #2 (Public review):

Summary:

The paper introduces the IBEX Knowledge-Base (KB), a shared online resource designed to help scientists working with immunofluorescence imaging. It acts as a central hub where researchers can find and share information about reagents, protocols, and imaging methods. The KB is not static like traditional publications; instead, it evolves as researchers contribute new findings and refinements. A key highlight is that it includes results of both successful and unsuccessful experiments, helping scientists avoid repeating failed experiments and saving time and resources. The platform is built on open-access tools ensuring that the information remains available to everyone. Overall, the KB aims to collaboratively accelerate research, improve reproducibility, and reduce wasted effort in imaging experiments.

Strengths:

(1) The IBEX KB is built entirely on open-source tools, ensuring accessibility and long-term sustainability. This approach aligns with FAIR data principles and ensures that the KB remains adaptable to future advancements.

(2) The KB also follows strict data organization standards, ensuring that all information about reagents and protocols is clearly documented and easy to find with little ambiguity.

(3) The KB allows scientists to report both positive and negative results, reducing duplication of effort and speeding up the research process.

(4) The KB is helpful for all researchers, but even more so for scientists in resource-limited settings. It provides guidance on finding affordable alternatives to expensive or discontinued reagents, making it easier for researchers with fewer resources to perform high-quality experiments.

(5) The KB includes a community discussion forum where scientists can ask for advice, share troubleshooting tips, and collaborate with others facing similar challenges.

Weaknesses:

(1) The potential impact of IBEX KB is very clear. However, the paper would benefit by also discussing more on KB maintenance and outreach, and how higher participation could be incentivized.

(2) Use of resources like GitHub may limit engagement from non-coding members of the scientific community. Will there be alternative options like a user-friendly web interface to contribute more easily?

Reviewer #3 (Public review):

Summary:

The authors have developed an interactive knowledge-base that uses crowdsourcing information on antibodies and reagents for immunofluorescence imaging.

Strengths:

The authors provide an extremely relevant and needed interphase for a community-based IF reagent and protocol knowledgebase, and a well-built interface. All the links on their website work, the information provided, reagents, datasets, videos, and protocols are very informative. The instructions for the community researchers to contribute are clear and they provide detailed instructions on how to technically proceed.

Weaknesses:

Reporting of the validation of antibodies could be improved. To increase public participation they suggest reducing the amount of details that one needs to submit to claim that something does not work. However, in our experience, this information is critical to be shared with the community.