A Novel Microchip Technique for Quickly Identifying Nanogranules in an Aqueous Solution by Transmission Electron Microscopy: Imaging of Platelet Granules
, Jungshan Chang 1,2, Wei-An Chen 3, Chih-Chun Chen 3, Hui-Min Chen 3, Chao-Chien Chang 4,5,6,* and Tsorng-Harn Fong 3,*
Round 1
Reviewer 1 Report
The authors reported the use of a novel microchip technique in order to identify native platelet granules. This is an interesting approach and it is less time consuming compared with the alternatives (Conventional TEM, Cryo-Electron Microscopy). Observing the platelet granules in aqueous solution is an advantage because the original ultrastructure is preserved and artifacts are avoided.
The results presented in the manuscript and obtained using negative staining and immunoelectron microscopy clearly demonstrate that this technique is well suited to identify platelet granules.
There are some small suggestions which can improve the manuscript.
It would the helpful for the reader if different types of platelet granules mentioned in the caption of Figure 3 (A) would be indicated using arrows, arrowheads, asterisks like in the other presented images.
Lines 204-205: The phrase: "The average diameter of granules which decorated by abundant gold particles was about 200 nm, which should be the α-granules (Fig. 4B)." should be modified. Example: "The average diameter of granules which were decorated by abundant gold particles was about 200 nm and they should be the α-granules (Fig. 4B)."
The authors should mention in Section 2.4 and 3.1 that the sample chamber is sealed at both ends with epoxy resin.
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Reviewer 2 Report
The authors have used a novel microchip technology to visualize sub cellular components in platelets. They isolated platelet granules and imaged them using the K-kit technology and verified further with antibody mediated staining. This technology is just another way of visualizing granules but it is expensive and requires special techniques that makes it highly unlikely to be used in every day also in research. The resolution obtained using this method cannot be used for any kind of qualitative or quantitative analysis. The authors do not provide any use and how it improves on the existing used technologies.
It will be of benefit if the authors can show that this technique is of relevance and use in platelet disorder.
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Reviewer 3 Report
The short paper by Trang et al. describes a quick and easy method to observe microvesicles obtained by platelets lysis using a commercially available microchip/chamber. The paper is well written and the subject worth the publication but the paper is poor in results and it should be considered as a short communication or a technical paper. Moreover microvesicles are more difficult to observe in body fluids where their concentration is low compared to the lysis model described. The immunoistochemistry used is correct even if exosomes should have been also identified.
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