Detection of large extracellular silver nanoparticle rings observed during mitosis using darkfield microscopy

Robert M. Zucker; Jayna Ortenzio; Laura L. Degn; William K. Boyes

doi:10.1371/journal.pone.0240268

Peer Review History

Original SubmissionJune 22, 2020
22 Jun 2020 Author Response Attachments Attachment Submitted filename: Rebuttle letter PLOS 1.docx https://doi.org/10.1371/journal.pone.0240268.r001
20 Jul 2020 Decision Letter - Amitava Mukherjee, Editor PONE-D-20-19210 Detection of Large Extracellular Silver Nanoparticle Rings observed during Mitosis using Darkfield Microscopy PLOS ONE Dear Dr. Zucker, Thank you for submitting your manuscript to PLOS ONE. After careful consideration, we feel that it 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 addresses the points raised during the review process. Please submit your revised manuscript by Sep 03 2020 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: http://journals.plos.org/plosone/s/submission-guidelines#loc-laboratory-protocols We look forward to receiving your revised manuscript. Kind regards, Amitava Mukherjee, ME, Ph.D. Academic Editor PLOS ONE Journal Requirements: When submitting your revision, we need you to address these additional requirements. 1. Please ensure that your manuscript meets PLOS ONE's style requirements, including those for file naming. The PLOS ONE style templates can be found at https://journals.plos.org/plosone/s/file?id=wjVg/PLOSOne_formatting_sample_main_body.pdf and https://journals.plos.org/plosone/s/file?id=ba62/PLOSOne_formatting_sample_title_authors_affiliations.pdf [Note: HTML markup is below. Please do not edit.] 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: Partly ******** 2. Has the statistical analysis been performed appropriately and rigorously? Reviewer #1: No ****** 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 ****** 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 ****** 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: PLOS ONE Detection of Large Extracellular Silver Nanoparticle Rings observed during Mitosis using Darkfield Microscopy Manuscript Number: PONE-D-20-19210 Article Type: Research Article In this article describe a technique that allows particles to be observed that would normally be invisible using other microscopic contrast techniques. The authors incorporated earlier suggestions by reviewers. There still seems to be scope of further improvement. The authors can go through the following points and revise the article. 1. Going through the abstract, the article and the conclusion, the authors have emphasised on the unusual appearance of silver nanoparticles around the cells undergoing mitosis. The authors leave the readers lost mentioning that these could be useful for future investigations. Perhaps the authors could emphasise more on this utility they foresee, thus, contributing to the scientific community. A histogram would definitely be a desired add-on. 2. The size of the nanoparticles in relation to the stage of the mitosis- can they be tabulated to give a rough idea to gauge the stage of cell division from the particle size, approximately? 3. Perhaps, some more data to establish the technique could be provided. 4. Also, the authors could go through the manuscript once again and remove additional texts. ****** 6. 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 [NOTE: If reviewer comments were submitted as an attachment file, they will be attached to this email and accessible via the submission site. Please log into your account, locate the manuscript record, and check for the action link "View Attachments". If this link does not appear, there are no attachment files.] While revising your submission, please upload your figure files to the Preflight Analysis and Conversion Engine (PACE) digital diagnostic tool, https://pacev2.apexcovantage.com/. PACE helps ensure that figures meet PLOS requirements. To use PACE, you must first register as a user. Registration is free. Then, login and navigate to the UPLOAD tab, where you will find detailed instructions on how to use the tool. If you encounter any issues or have any questions when using PACE, please email PLOS at figures@plos.org. Please note that Supporting Information files do not need this step. https://doi.org/10.1371/journal.pone.0240268.r002
Revision 1
21 Aug 2020 Author Response Reviewer #1: PLOS ONE Detection of Large Extracellular Silver Nanoparticle Rings observed during Mitosis using Darkfield Microscopy Manuscript Number: PONE-D-20-19210 Article Type: Research Article Dear Dr Mukherjee , Thank you for giving us an opportunity to revise our manuscript, to answer ther reveiwers concens and make it more comprehnsible for the PLOS1 audinece. We have addressed your concerns and hope it is now suitable for publication in PLOS 1. Comment: There still seems to be scope of further improvement. The authors can go through the following points and revise the article. 1. Going through the abstract, the article and the conclusion, the authors have emphasized on the unusual appearance of silver nanoparticles around the cells undergoing mitosis. The authors leave the readers lost mentioning that these could be useful for future investigations. Perhaps the authors could emphasize more on this utility they foresee, thus, contributing to the scientific community. A histogram would definitely be a desired add-on. Response: An explanation for how the nanoparticles rings originate during prophase is presented in the discussion and mentioned in the abstract. A new histogram (Figure 5) of ring sizes has been added as suggested by the reviewer to summarize the sizes of rings observed. An interpretation of the formation and thickness of the rings occurring during the prophase stage of mitosis has been proposed in the discussion. Additional data on the size of the rings and their significance have been provided in the discussion. The ring structure may be integrated with events that occur during prophase as the cell transitions between G2 restriction point and metaphase. The relationship of these ring structures appears to be another useful parameter to describe the events between prophase and metaphase which may correlate to the other events involving microtubules and chromosome condensation. 2. The size of the nanoparticles in relation to the stage of the mitosis- can they be tabulated to give a rough idea to gauge the stage of cell division from the particle size, approximately Response. The nanoparticles were not observed to be different during the stage of the cell cycle or mitosis. Particles inside the cell are brighter and thus considered larger and clumped compared to particles outside the cell or in the rings. The measurement of nanoparticles by darkfield microscopy was found to be about 0.37 μm. The apparent size of the nanoparticles inside the cell are clumped and appear to be larger at 0.61 μm. These data are incorporated into the text of the manuscript. It should be emphasized that dark field microscopy does not measure the actual size of the object or particle but shows the reflection of light from the location of the particle. The darkfield observation of Ag nanoparticles show the particles to appear bigger than their actual physical size. The rings consist of particles that appear to have similar sizes to the extracellular particles distributed on the slide surface. It was not possible to measure the individual size of particles in the rings due to their high density, clumping and their location in a 3d structure. 3. Perhaps, some more data to establish the technique could be provided. Response: A histogram of the rings structures was provided in in a new Figure 5. We also measured the size of the nanoparticles using Nikon elements by darkfield imaging. It was found that these particles have an apparent size of 0.37 μm by dark field microscopy, through measuring the area of reflected light as opposed to actual physical size of the particle which was described as 80 nm by the manufacturer. The darkfield methodology can show particles below the diffraction limit of the microscope. The resulting image creates a spot that is larger than its physical size. 4. Also, the authors could go through the manuscript once again and remove additional texts. Response –The text has been revised and additional text removed. Attachments Attachment Submitted filename: RESPONSE TO REVIEWER ARTICLE.odt https://doi.org/10.1371/journal.pone.0240268.r003
8 Sep 2020 Decision Letter - Amitava Mukherjee, Editor PONE-D-20-19210R1 Detection of Large Extracellular Silver Nanoparticle Rings observed during Mitosis using Darkfield Microscopy PLOS ONE Dear Dr. Zucker, Thank you for submitting your manuscript to PLOS ONE. After careful consideration, we feel that it 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 addresses the points raised during the review process. Please submit your revised manuscript by Oct 23 2020 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: http://journals.plos.org/plosone/s/submission-guidelines#loc-laboratory-protocols We look forward to receiving your revised manuscript. Kind regards, Amitava Mukherjee, ME, Ph.D. Academic Editor PLOS ONE [Note: HTML markup is below. Please do not edit.] 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 ******** 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: Partly ****** 3. Has the statistical analysis been performed appropriately and rigorously? Reviewer #1: 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 ****** 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: No ****** 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 have answered all the queries as asked. However, the way of writing of the article needs to be checked further. Its not lucid and hence, reading the article doesnot garner much interest.It would be nice if the authors could go through again and revise the language and the style of writing as per the journal standards. ****** 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 [NOTE: If reviewer comments were submitted as an attachment file, they will be attached to this email and accessible via the submission site. Please log into your account, locate the manuscript record, and check for the action link "View Attachments". If this link does not appear, there are no attachment files.] While revising your submission, please upload your figure files to the Preflight Analysis and Conversion Engine (PACE) digital diagnostic tool, https://pacev2.apexcovantage.com/. PACE helps ensure that figures meet PLOS requirements. To use PACE, you must first register as a user. Registration is free. Then, login and navigate to the UPLOAD tab, where you will find detailed instructions on how to use the tool. If you encounter any issues or have any questions when using PACE, please email PLOS at figures@plos.org. Please note that Supporting Information files do not need this step. https://doi.org/10.1371/journal.pone.0240268.r004
Revision 2
19 Sep 2020 Author Response Re: PONE-D-20-19210R1 Amitava Mukherjee, ME, Ph.D. Academic Editor PLOS ONE Dear Dr Mukherjee, Thank you and your peer reviewers for the careful evaluation of our manuscript entitled: “Detection of Large Extracellular Silver Nanoparticle Rings observed during Mitosis using Darkfield Microscopy”. For our previous revision, the reviewers indicated that the revised manuscript had addressed all the technical issues identified, but that the writing style and editorial quality were insufficient. The reviewers, however, did not identify any specific sections of the manuscript that needed attention. Today, we are submitting a revised version of the manuscript which has been thoroughly revised to improve the editorial quality. Among other changes, we have removed redundancies, simplified and corrected sentence structures, corrected grammar, improved the consistency of acronyms and symbols, and corrected reference styles. We submit a “marked” version in which changes to the manuscript are indicated using the MS Word “track change” feature. You will see that the editorial revisions were extensive. We also submit a “clean” version in which all the changes were accepted. Overall, we believe, the quality and readability of the manuscript were improved, which should make it more interesting for the readers of PLOS ONE. We hope that your reviewers will observe the improved quality of the manuscript and determine that it is now acceptable for publication in PLOS ONE. Sincerely, Robert Zucker, PhD U.S. Environmental Protection Agency Research Triangle Park, NC 27701 Email: zucker.robert@epa.gov Attachments Attachment Submitted filename: Response to reviewers.docx https://doi.org/10.1371/journal.pone.0240268.r005
23 Sep 2020 Decision Letter - Amitava Mukherjee, Editor Detection of Large Extracellular Silver Nanoparticle Rings observed during Mitosis using Darkfield Microscopy PONE-D-20-19210R2 Dear Dr. Zucker, 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. An invoice for payment will follow shortly after the formal acceptance. To ensure an efficient process, please log into Editorial Manager at http://www.editorialmanager.com/pone/, click the 'Update My Information' link at the top of the page, and double check that your user information is up-to-date. If you have any billing related questions, please contact our Author Billing department directly at authorbilling@plos.org. If your institution or institutions have a press office, please notify them about your upcoming paper to help maximize its impact. If they’ll be preparing press materials, please inform our press team as soon as possible -- no later than 48 hours after receiving the formal acceptance. Your manuscript will remain under strict press embargo until 2 pm Eastern Time on the date of publication. For more information, please contact onepress@plos.org. Kind regards, Amitava Mukherjee, ME, Ph.D. Academic Editor PLOS ONE Additional Editor Comments (optional): Reviewers' comments: https://doi.org/10.1371/journal.pone.0240268.r006
Formally Accepted
8 Oct 2020 Acceptance Letter - Amitava Mukherjee, Editor PONE-D-20-19210R2 Detection of large extracellular silver nanoparticle rings observed during mitosis using darkfield microscopy Dear Dr. Zucker: I'm pleased to inform you that your manuscript has been deemed suitable for publication in PLOS ONE. Congratulations! Your manuscript is now with our production department. If your institution or institutions have a press office, please let them know about your upcoming paper now to help maximize its impact. If they'll be preparing press materials, please inform our press team within the next 48 hours. Your manuscript will remain under strict press embargo until 2 pm Eastern Time on the date of publication. For more information please contact onepress@plos.org. If we can help with anything else, please email us at plosone@plos.org. Thank you for submitting your work to PLOS ONE and supporting open access. Kind regards, PLOS ONE Editorial Office Staff on behalf of Professor Dr. Amitava Mukherjee Academic Editor PLOS ONE https://doi.org/10.1371/journal.pone.0240268.r007

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