PONE-D-22-20783Imaging breast malignancies with the Twente Photoacoustic Mammoscope 2PLOS ONE

Dear Dr. Schoustra,

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Reviewers' comments:

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Comments to the Author

1. Is the manuscript technically sound, and do the data support the conclusions?

The manuscript must describe a technically sound piece of scientific research with data that supports the conclusions. Experiments must have been conducted rigorously, with appropriate controls, replication, and sample sizes. The conclusions must be drawn appropriately based on the data presented.

Reviewer #1: Partly

Reviewer #2: Yes

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2. Has the statistical analysis been performed appropriately and rigorously?

Reviewer #1: N/A

Reviewer #2: Yes

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3. Have the authors made all data underlying the findings in their manuscript fully available?

The PLOS Data policy requires authors to make all data underlying the findings described in their manuscript fully available without restriction, with rare exception (please refer to the Data Availability Statement in the manuscript PDF file). The data should be provided as part of the manuscript or its supporting information, or deposited to a public repository. For example, in addition to summary statistics, the data points behind means, medians and variance measures should be available. If there are restrictions on publicly sharing data—e.g. participant privacy or use of data from a third party—those must be specified.

Reviewer #1: Yes

Reviewer #2: Yes

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Reviewer #1: Yes

Reviewer #2: Yes

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5. Review Comments to the Author

Please use the space provided to explain your answers to the questions above. You may also include additional comments for the author, including concerns about dual publication, research ethics, or publication ethics. (Please upload your review as an attachment if it exceeds 20,000 characters)

Reviewer #1: The authors reported clinical measurements of breast cancer patients with the 3D Twente photoacoustic mammoscope prototype. Specifically, 30 patients in total were scanned and 4 out of 19 malignant cases were closely investigated. Photoacoustic imaging results were qualitatively compared with craniocaudal X-ray images, ultrasound images, and MRI images. Even though a larger cohort is expected for more convincing conclusions, the scientific findings of the four cases are of interest to this journal. The following aspects can be improved before further consideration of publication:

1. One caveat of the study is that there is no control group. It is unclear how different images will look like in benign cases, especially in PA images. Authors should at least analyze some benign cases and show the X-ray, MRI, US, and PA images for comparison.

2. The authors claim that “to enhance signals coming from the deeper vessels, adaptive intensity modulation was applied”. However, from the justification as well as results shown in figures (such as Fig 6), it seems that adaptive intensity modulation does not compensate for low illumination in deep regions. Instead, it adjusts image intensity with respect to local variability, which is more anatomy dependent rather than depth dependent. Please elaborate or correct.

3. To compensate for depth dependent PA intensity, optical fluence compensation is necessary, which is missing in this work. There have been multiple works on optical fluence compensation for breast. Suggest authors at least implementing a simple model-based method to consider exponentially attenuated light fluence given the boundary of the breast.

4. Also in image processing, why are the final images obtained by summing the adaptive modulated image and the median filtered image, instead of applying the adaptive modulation directly on the filtered image?

5. It is difficult to correlate different modalities without registration. The authors discussed the difficulty of cross-modality registration due to a lack of anatomical landmarks, but also mentioned a previous work where MRI and PA images are deformably registered and fused in reference [7] in Introduction. Why not applying the same technique to register at least MR and PA images?

Other comments:

6. In Materials and Methods, the authors mentioned that two US probes were used in the study, i.e. an L12-5 50 and an L18-5. However, only one ultrasound image was shown for each case in Results. Please elaborate the difference of the images acquired from the two probes, and how they were used in diagnosis.

7. How does the entropy map look like for the other three cases?

8. In discussion, the authors explained that the oxygen saturation was not evaluated due to the difference in illuminated tissue with the two wavelengths. This is confusing, why would the illuminated tissue change at different wavelengths? Please clarify.

9. The discussion of improvements achieved from this work compared with the previous generation of Twente mammoscope [5, 6] is inadequate.

10. The discussion can be organized more systematically and concisely by concluding all cases by their features, instead of listing one by one again as in Results. Suggest authors using “can be” instead of “might be” or “seems that” to conject.

Reviewer #2: The manuscript verified the clinical feasibility of PAM-02 system to diagnose breast cancer. Overall the manuscript is acceptable to publication. Please address the minor question of the PAM-02 system.

- The authors used 795nm and 1064nm lasers for imaging. Please justify why both wavelength are chosen. and other research (10.1038/s41467-018-04576-z) stated single 1064 laser was used to detect breast cancer, why did you use dual-band laser?

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Reviewer #1: No

Reviewer #2: No

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PONE-D-22-20783R1Imaging breast malignancies with the Twente Photoacoustic Mammoscope 2PLOS ONE

Dear Dr. De Santi,

Thank you for submitting your manuscript to PLOS ONE. After careful consideration, we feel that minor revision would make your article fully satisfy PLOS ONE’s publication criteria. Therefore, we invite you to submit a revised version of the manuscript that addresses the minor points raised during the review process.

Please submit your revised manuscript by Jan 13 2023 11:59PM. If you will need more time than this to complete your revisions, please reply to this message or contact the journal office at plosone@plos.org. When you're ready to submit your revision, log on to https://www.editorialmanager.com/pone/ and select the 'Submissions Needing Revision' folder to locate your manuscript file.

Please include the following items when submitting your revised manuscript:

• A rebuttal letter that responds to each point raised by the academic editor and reviewer(s). You should upload this letter as a separate file labeled 'Response to Reviewers'.
• A marked-up copy of your manuscript that highlights changes made to the original version. You should upload this as a separate file labeled 'Revised Manuscript with Track Changes'.
• An unmarked version of your revised paper without tracked changes. You should upload this as a separate file labeled 'Manuscript'.

If you would like to make changes to your financial disclosure, please include your updated statement in your cover letter. Guidelines for resubmitting your figure files are available below the reviewer comments at the end of this letter.

If applicable, we recommend that you deposit your laboratory protocols in protocols.io to enhance the reproducibility of your results. Protocols.io assigns your protocol its own identifier (DOI) so that it can be cited independently in the future. For instructions see: https://journals.plos.org/plosone/s/submission-guidelines#loc-laboratory-protocols. Additionally, PLOS ONE offers an option for publishing peer-reviewed Lab Protocol articles, which describe protocols hosted on protocols.io. Read more information on sharing protocols at https://plos.org/protocols?utm_medium=editorial-email&utm_source=authorletters&utm_campaign=protocols.

We look forward to receiving your revised manuscript.

Kind regards,

Jeeun Kang, Ph.D.

Academic Editor

PLOS ONE

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Please review your reference list to ensure that it is complete and correct. If you have cited papers that have been retracted, please include the rationale for doing so in the manuscript text, or remove these references and replace them with relevant current references. Any changes to the reference list should be mentioned in the rebuttal letter that accompanies your revised manuscript. If you need to cite a retracted article, indicate the article’s retracted status in the References list and also include a citation and full reference for the retraction notice.

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Reviewers' comments:

Reviewer's Responses to Questions

Comments to the Author

1. If the authors have adequately addressed your comments raised in a previous round of review and you feel that this manuscript is now acceptable for publication, you may indicate that here to bypass the “Comments to the Author” section, enter your conflict of interest statement in the “Confidential to Editor” section, and submit your "Accept" recommendation.

Reviewer #1: (No Response)

Reviewer #2: All comments have been addressed

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2. Is the manuscript technically sound, and do the data support the conclusions?

The manuscript must describe a technically sound piece of scientific research with data that supports the conclusions. Experiments must have been conducted rigorously, with appropriate controls, replication, and sample sizes. The conclusions must be drawn appropriately based on the data presented.

Reviewer #1: Yes

Reviewer #2: Yes

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3. Has the statistical analysis been performed appropriately and rigorously?

Reviewer #1: Yes

Reviewer #2: Yes

**********

4. Have the authors made all data underlying the findings in their manuscript fully available?

The PLOS Data policy requires authors to make all data underlying the findings described in their manuscript fully available without restriction, with rare exception (please refer to the Data Availability Statement in the manuscript PDF file). The data should be provided as part of the manuscript or its supporting information, or deposited to a public repository. For example, in addition to summary statistics, the data points behind means, medians and variance measures should be available. If there are restrictions on publicly sharing data—e.g. participant privacy or use of data from a third party—those must be specified.

Reviewer #1: Yes

Reviewer #2: Yes

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5. Is the manuscript presented in an intelligible fashion and written in standard English?

PLOS ONE does not copyedit accepted manuscripts, so the language in submitted articles must be clear, correct, and unambiguous. Any typographical or grammatical errors should be corrected at revision, so please note any specific errors here.

Reviewer #1: Yes

Reviewer #2: Yes

**********

6. Review Comments to the Author

Please use the space provided to explain your answers to the questions above. You may also include additional comments for the author, including concerns about dual publication, research ethics, or publication ethics. (Please upload your review as an attachment if it exceeds 20,000 characters)

Reviewer #1: Partially addressed:

Comment 1: Let alone the benign cases for now, is it possible to show images of normal breasts so that readers can have a clearer sense how the vascular features are different in breast tumors from normal breasts?

Suggestions:

Comments 2, 3, 4, 5, 7 are well addressed. If the authors choose to publish the peer review history then it might be fine. Otherwise, the revised manuscript is a bit too simplified. If the authors choose not to publish the peer review history, suggest authors include the analysis and justifications that you responded to the rebuttal to the Discussion Section to make the manuscript more comprehensive. Figures R2, R3, R5, R7 should be added to Supporting Information. Figure R4 should be added to Methods.

Reviewer #2: (No Response)

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7. PLOS authors have the option to publish the peer review history of their article (what does this mean?). If published, this will include your full peer review and any attached files.

If you choose “no”, your identity will remain anonymous but your review may still be made public.

Do you want your identity to be public for this peer review? For information about this choice, including consent withdrawal, please see our Privacy Policy.

Reviewer #1: No

Reviewer #2: No

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Attachments

Attachment

Submitted filename: PONE-D-22-20783_YWu_R2.docx

Imaging breast malignancies with the Twente Photoacoustic Mammoscope 2

PONE-D-22-20783R2

Dear Dr. De Santi,

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Academic Editor

PLOS ONE