Development of a Single-Piece Sperm Counting Chamber (SSCC) for Aquatic Species
, Yue Liu 1,2, Elizabeth L. Reich 1, Jason Eades 1, Terrence R. Tiersch 2 and William Todd Monroe 1,*Round 1
Reviewer 1 Report
Dear authors,
The manuscript is well written; clear, precise and easy to understand. I felt in love for your schematic diagram and the SSCC prototype. Please, considered patenting your idea (if it is possible in your country). A very large amount of work was involved in the study, and as far as I can determine, the work is solid. In my opinion, the manuscript fully deserves to be published.
By the way, I detected some little mistakes. Please, check:
. Simple summary: page 1, line 16 – change “protypes” to “prototypes”
. M&M: page 3, lines 114-116 – check the right order of the Figures legends, in my opinion should be Figure 2E at line 114, Figure 2C at line 115 and Figure 2D at line 116
. M&M: page 6, line 244 – change “ANVA” to “ANOVA”
. Results: page 7, lines 287-289 – Rephrase the sentence: “The open-midpoint design was superior to the enclosed chamber and closed grid designs but was significantly lower (P = 0.0301 for 1 × 108) for the 1 × 108 cells/mL counts.”, it sounds clumsy.
. Results: page 8, lines 301 – at “Figure 3”, add “, top”
. Results: page 8, lines 306 – at the end, add “(Figure 3, bottom)”
Figures captions
. Figure 3: page 8, line 308 – change “goldish” to “Gold fish”
. Figures 3-5: change “108” to “108”
Have a nice day!
Author Response
Please see attached.
Author Response File: 
Author Response.pdf
Reviewer 2 Report
General comments:
The authors describe the development of a prototype single-piece sperm counting chamber (SSCP) for aquatic species, and the differences in accuracy of multiple grid designs, their accuracy when used repeatedly, and compared to existing products in this paper. The results presented in this manuscript are very clear and convincing. The authors have established a flexible SSCP manufacturing protocol that is cheaper to produce than the conventional devices and can be adjusted to a grid height suitable for the size of sperm cells of various aquatic species. The authors' claim that such a product would assist sperm cryopreservation and reproductive protocols in aquatic species is agreed upon. This reviewer considers that this manuscript is in line with the purpose of Fishes and worthy of publication. However, I believe the points commented below are open to revision, and I would ask the authors to reconsider them to improve this manuscript.
Specific comments:
- Figures 1 and 2 should be reordered. Also, the original Figure 1 should not be inserted at the end of the Introduction, but in the M&M section, where it would be more appropriate.
- The original Figure 2 does not match the description in the text
L113: “Figure 1b” -> “Figure 2b”
L114: “Figure 2c” -> “Figure 2e”
L115: “Figure 2d” -> “Figure 2c”
L116: “Figure 2e” -> “Figure 2c”
Figure 2a should show a 5 x 5 pattern without walls in contrast to 2b. Alternatively, the text description and caption should be rewritten to match the figure.
- The first paragraph of the Results section (L250-L263) contains much of what should be in the M&M section.
- For Figuire3,4,5,6, the title of the vertical axis is "Cell concentration (x 10^8 cells/ml)" and the values are “0, 0.5, 1, ...”. It is easier to see. In addition, the caption of the figure usually does not include the results ; such as “Enclosed wall design concentrations were significantly higher ….” These should be removed.
Author Response
Please see the attachment.
Author Response File: Author Response.pdf
