Automated Immunomagnetic Enrichment and Optomicrofluidic Detection to Isolate Breast Cancer Cells: A Proof-of-Concept towards PoC Therapeutic Decision-Making

Round 1

Reviewer 1 Report

The manuscript demonstrated an automated workflow for immunomagnetic enrichment and microfluidic detection which is helpful in diagnosis and personalized treatment of breast tumors. It seems that this automated workflow have been reported previously. This manuscript should be less innovative than the previous paper describing its design. However, considering the future prospects in application, and some of the methodological conditions in the paper are useful for reference, I suggest it be accepted. 

Though, the authors need consider the following questions.

1, Since this article is published independently, it is difficult for readers to search the previous paper, a general introduction about the framework or design of the automated system is needed.

2, This research mainly carried out some optimization work. Personally, some of these optimizations are questionable significance and not innovative, eg.

In part “2.3. Establishment of a rapid protocol to assay cancer-related mRNAs……”, one-step qPCR was performed with a commercial one-step kit, the SensiFAST™ Probe No-ROX One-Step kit (Meridian Bioscience, Luckenwalde, DE). The qPCR was conducted in a separate cycler, (BioRad CFX96 Touch Real-Time PCR Detection System, Feldkirchen, DE), which is a follow-up procedural experiment but not an approach established by this study. As the simplified protocol claimed by the authors, reverse transcription was extended to 20 minutes and extension time was set to 10 seconds, is just a simple setup. Such similar time setting is often used for similar experiments. What's more, this paper also found that under the same condition, the efficacy of one-step method is not as good as that of two-step, so the one-step method with limited simplicity has not so much significance. Moreover, this paragraph seems unrelated with the topic ‘Automated immunomagnetic enrichment and optomicrofluidic……’, thus not worth reporting in a large section.

3, In line 4, PoC is not a well-known acronym, and the full name should be given where appropriate in the text.

4, In line 41, “cancer entity” seems unlike a normative medical word. Do the authors mean the type of solid tumor?

5, Line 217-220, wasn’t it caused by the proliferation of single cells in the culture dish that Ct value (34.3±1.5) of the single cells in the culture well less than that of the suspension cells (37.5 ± 1.6)? Please explain.

Author Response

Please see the attachment.

Author Response File: Author Response.pdf

Reviewer 2 Report

The manuscript “Automated immunomagnetic enrichment and optomicrofluidic detection to isolate breast cancer cells: A proof-of-concept towards PoC therapeutic decision-making” by Stiefel et al. describes the use of a microfluidic cell for immunomagnetic enrichment of cancer cells.

The literature is well-reviewed and represents the state-of-the-art. However, the authors should add a further paragraph on existing cell-enrichment processes in the introduction.

The methods are well explained and allow for reproducibility. The results are nicely discussed. However, also in the discussion a qualitative comparison between your results and existing systems would significantly improve the quality of the manuscript.

I only have a few minor issues which need to be addressed:

Figure 1: the font is very small in the figures and an enlargement helps to better understand and read the figures.

Author Response

Please see the attachment.

Author Response File: Author Response.pdf