PONE-D-24-00790Scalable lipid droplet microarray fabrication, validation, and screeningPLOS ONE

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Academic Editor

PLOS ONE

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Additional Editor Comments :

Thank you for submitting your manuscript to PLOS ONE. After careful consideration, we feel that the work is timely, and the manuscript has merit but does not fully meet PLOS ONE’s publication criteria as it currently stands. Therefore, we invite you to submit a revised version of the manuscript that could address the points raised by all the reviewers. The manuscript requires a major revision prior to a final decision.

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Reviewers' comments:

Reviewer's Responses to Questions

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: Yes

Reviewer #2: Yes

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

Reviewer #1: Yes

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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4. 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

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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: In this manuscript, authors have fabricated liquid droplet microarray for cellular validation and screening using nanointaglio. And, the authors have succeeded to screen the screening. From these achievements, this manuscript seems useful for high-throughput screening for cell analysis. However for publication of this manuscript, authors should reconsider the several points.

1. The stability and lifetime of droplets. Is these droplets can be printed on the substrate stably? If possible, the life time also should be discussed.

2. Are the cell density affect to the screening efficiency?

3. If possible, after the printing with different pressures, differences of sizes of droplet should be indicated using SEM imaging.

Reviewer #2: The work reported in the manuscript is interesting in terms of both the method to prepare droplets, and the cell responses to droplets. The former is the focus of the manuscript. In forming the droplets, the first step is quite ordinary, but it is clever to split the ink droplets by using intaglio stamp. I have the following comments.

1. Figure 2 may better show the images of the droplet array prepared under the optimal pressure of 16 N/m^2, along with current representative images. It is important to show 100% uniformity droplets.

2. What is the interfacial tension between aqueous medium and the lipid droplets? This tension determines whether the deformed droplets can readily resume a spherical cap shape.

3. The authors pointed out that the droplets are not stable as they are immersed into the aqueous medium. The reason for such instability is the development of four phase contact line, which is unavoidable in the process of bringing oil-like droplets into water, or vice verse. See the detailed mechanism: Splitting droplets through coalescence of two different three-phase contact lines. Soft matter 15 (30), 6055-6061 (2019). The approach taken by the authors to improve the stability of the droplets is ok. An alternative way may be to improve the viscosity of the droplets, or control the speed for immersion. The authors may provide these possibilities in their outlook for future work.

4. The surface gettability of the stamps would play an important role in precisely distributing the droplet liquid. What are the advancing and receding contact angles of the stamp materials?

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Reviewer #1: No

Reviewer #2: Yes: Xuehua Zhang

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Scalable lipid droplet microarray fabrication, validation, and screening

PONE-D-24-00790R1

Dear Dr. Lenhert,

We’re pleased to inform you that your manuscript has been judged scientifically suitable for publication and will be formally accepted for publication once it meets all outstanding technical requirements.

Within one week, you’ll receive an e-mail detailing the required amendments. When these have been addressed, you’ll receive a formal acceptance letter and your manuscript will be scheduled for publication.

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Kind regards,

Jian Xu, Ph.D.

Academic Editor

PLOS ONE

Additional Editor Comments (optional):

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: All comments have been addressed

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

**********

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: The authors were able to accurately reconsider the points raised by the reviewers. Revisions are satisfactory. Hence, this manuscript is able to publish in PLOS ONE.

Reviewer #2: The authors have made careful revision. All the comments haven been addressed adequately. I have no more comments.

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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: Yes: Tatsuro Endo

Reviewer #2: No

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