PONE-D-23-30649A comparative study of blood cell count in four automated hematology analyzers: an evaluation of the impact of preanalytical factorsPLOS ONE

Dear Dr. Åstrand,

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Enoch Aninagyei, PhD

Academic Editor

PLOS ONE

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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: I Don't Know

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: The authors compare 4 hematology Analyzers in whole cell blood count, but most of clinical laboratory has one or two analyzer that they are working.

The authors neither experiment nor discuss about RBC, HCt, Hgb and RBC index's. may be they will do it in their next manuscript.

the aim of this study is not clear.

Reviewer #2: Well written and well formatted article. Technically sound.Excellent work.

The methodology is sound,However the sample size could have been higher to establish more conclusive findings.This helps to identify possible pre analytical variables and their influence on outcomes

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Reviewer #1: No

Reviewer #2: Yes: Bobby Abraham

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PONE-D-23-30649R1A comparative study of blood cell count in four automated hematology analyzers: an evaluation of the impact of preanalytical factorsPLOS ONE

Dear Dr. Åstrand,

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 Mar 14 2024 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,

Enoch Aninagyei, PhD

Academic Editor

PLOS ONE

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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 #3: (No Response)

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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 #3: Partly

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

Reviewer #3: Yes

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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 #3: No

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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 #3: No

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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 #3: The authors have compared the counts of white blood cells in different hematology platforms in terms of:

1- Storage temperature

2- Time to analysis

3- Mechanical stress

With later a special focus on eosinophils.

This comparative study is needed to provide insight into the factors that contribute to the variability of results across platforms.

There are a number of concerns, outlined below, that need to be addressed in order to publish this to be of use to the scientific and medical community.

1- The authors need to justify the focus on eosinophils. Perhaps stating in the introduction.

2- The experimental design is a little confusing and hard to follow. I suggest adding a flow chart of the different participants and their characteristics and different experiments and manipulations done.

3- Please define what you mean by baseline in the methods section.

4- In line 109 it is mentioned that two tubes from each donor were put in 4 different temperature. Were each tube split into two different tubes in order to have them in 4 different temperatures? This is not clear. There are many instances where the methodology of handling the samples is not clear. Therefore, it will be hard for other scientists to reproduce the protocol used.

5- In line 122, RT, which I assume is refereeing to room temperature, was not defined prior to this point

6- I have a concern regarding patch effects of the different experiments run at different places and different times and yet they were compared to each other in the results. Is there a way statistically to control for that?

7- In Line 133, “(w/wo)” means with or without atopy? Is there a reason for not using one, either with or without, or using both groups separately; so the healthy will have n=6 (3+3).

8- For the flow cytometry and EDN assay, were there technical replicates? If not please justify.

9- In line 167, it is indicated that the diluted samples had an upper range of 640 ng/ml which is outside the standard curve. It is not very clear this section in the methods as there are many different brackets and they are not always properly used and effects the understanding the sentences. This is seen all-over the manuscript and needs revisions.

10- The results section is disorganized and very hard to follow. it needs to be organized with clear subheadings and better flow. For example, in the first section of the results, after the demographics of the participants, is not very clear what experiments our question is being answered especially that it feels like some the results are just repeated in other sections. Moreover, the description of the results between the different platforms and the different temperature and times need to be written in a more clear. For example, when the increase is mentioned by X (y-z) are you referring to a percent increase and a range. This need to be added in the statistical analysis section, where you mention that data are presented as ………. The results section needs major reorganization to help the readers follow the logic of the experiments.

11- The discussion need to describe the results in more details. For example, in line 358, the authors need to rewrite this to better descibe their findings in the different experiments. Also, in line 366, the authors state that leukocytes are sensitive to external temperatures; was this based on the their data or from the literature? They need to specify that.

12- Discuss the significance of examining eosinophils specifically.

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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 #3: Yes: Sameera Abuaish

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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.

A comparative study of blood cell count in four automated hematology analyzers: an evaluation of the impact of preanalytical factors

PONE-D-23-30649R2

Dear Dr. Annika Åstrand,

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.

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Kind regards,

Enoch Aninagyei, PhD

Academic Editor

PLOS ONE

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