Response: As the Reviewer's good instruction, we have tried our best to revise the English of the whole MS carefully. In order to make the whole MS better understanding, we have revised some
long sentences into short sentences and edited the whole MS according to the Reviewer’s instruction. Meanwhile, we also have asked some colleagues who are skilled authors of English language papers to help us for checking the English (see the revised MS). We hope that the language is now acceptable for the next review process. Special thanks to you for your good comments.
Reviewer 3#
1. Response to comment: English should be checked throughout the text by a native English speaker.
Response: According to the reviewer's good instruction, we have revised the whole manuscript
carefully and tried to avoid any grammar or syntax error. In addition, we have asked several
colleagues who are skilled authors of English language papers to check the English. We hope that
the language is now acceptable for the next review process.
2. Response to comment: The manuscript is too long. It must be shortened. The authors must be
more concise. The introduction takes three pages. In fact, it is very hard to read the paper. There are several sentences that should be changed for a better understanding. Some corrections are done in the margins of the manuscript (pods file). I enclosed a copy of that. Response: We agree the reviewer's good advice. Yes, the manuscript is too long (especially the
part of introduction), which is very hard to read the paper. And that, there are several sentences are hard for understanding. Thus, we have revised the original manuscript in order to reduce the
length of the manuscript and make it better understanding (especially the part of Introduction).
However, due to additional experiments and explanations are added in the revised manuscript
according to the other Reviewer′s suggestion, the revised manuscript is still long in some sort. Nevertheless, we have revised the sentences (especially some long sentences) for the whole manuscript in order to make the manuscript more concise. Especially, the corrections done in the margins of the manuscript (pods file) which the Reviewer enclosed are very helpful to us. We are very appreciated for the Reviewers good comments and corrections made for our manuscript.
3. Response to comment: Repetitions and several adjectives should be avoided. For example:
authors use \text.
Also, they indicate \SC become weaker when the time goes on thus, no Raman signal will be obtained for a long, long time. It means, that the time deteriorates Ag surface. I think that \a good adjective.
Response: It is really true as the Reviewer suggested that some repetitions and several adjectives should be avoided. Thus, we have made corrections according to the Reviewer’s good instructions. We have deleted the repeated words such as \\and \not studied the SERS spectra of SC for a long, long time. Thus, the using of \describe the SERS substrate of Ag nanofilm is inapposite. Considering the Reviewer’s good suggestion, we have deleted this adjective in some sentences of the revised manuscript.
4. Response to comment: About organization of the manuscript. There are too many epigrap****** in
section 2. I propose the following points:
2.1. Reagents. Preparation of ****** and ****** (old points 2.1 and 2.2 together)
2.2. Preparation of ******-protected Ag nanoparticles and ******s (old points 2.3 and 2.4
together)
2.3. Experimental equipments (old points 2.5; 2.6 and 2.7 together)
Response: Considering the Reviewer’s good suggestion, we have re-organized the epigrap****** in section 2. We have organized three parts for the section 2 of the revised manuscript. The epigrap****** in section 2 are as following: 2.1. Reagents and preparation of ****** and ******
2.2. Preparation of ******-protected ****** (******-Ag CNPs) and ******s 2.3. Experimental equipments
Special thanks to you for your good comments.
Dear Editors and Reviewers.
We have tried our best to revise and improve the manuscript and made great changes in the
manuscript according to the Reviwers′good comments. And here we did not list the changes but
marked in red in revised paper.
We appreciate for Editors/Reviewers’ warm work earnestly, and hope that the corrections will
meet with approval.
Once again, thank you very much for your comments and suggestions.
We look forward to your information about my revised papers and thank you for your good
comments.
Yours sincerely, R.M. Liu
5. Response to comment: Point 3.3 is entitled \and repeatability of ******s\Some
questions: Here, it is shown the dependence of SERS relative intensities of SC on time. Is it the same sample (******-adsorbate) that is stored and then SERS spectra are recorded at different times? or instead, the same ****** is stored and then the adsorbate is added at different times and thereafter the SERS spectra are recorded?? Authors should clarify the procedure. Are different results expected with these two procedures? Have been these two methods checked? It is possible that only ******s stored without adsorbate are active for a longer time?
Experimentally it is often observed that roughed surface in SERS
produces molecular degradation and a comparation between Raman and SERS spectra is necessary to identify the molecular fundamental modes. Why the authors did not record Raman spectra of SC? Raman spectra should be added to Fig. 5. The point 3.3 should be 3.2 because a characteristic of surface is explained and must follow point 3.1. The old 3.2 becomes 3.3. Moreover, old epigrap****** 3.2.1. and 3.2.2 should be removed and a unique point 3.3. should be presented instead and entitled \ty of ******s. SERS spectra of ****** and ******\Also, epigraph 3.4 must be removed and the text must follow to the new section 3.3.
Response: We are very sorry for our unclear report in the section of 3.3. For the first question proposed by the Reviewer, our answer is as following:
Yes, in the section of 3.3, the main intention is to display the stability and reproducibility of Ag
nanofilms. Fig. 6 (in the original manuscript, Fig. 5 in the revised manuscript) shows the dependence of SERS relative intensities of SC on time. In this section, the ******s are stored and then the adsorbate is added at different times and thereafter the SERS spectra are recorded. For the second question proposed by Reviewer, our answer is as following:
As the Reviewer's good instruction, we have clarified the procedure in the revised manuscript.
However, in this study, we have not studied the first procedure of the same sample (******-adsorbate) that is stored and then SERS spectra are recorded at different times. Thus, these two methods have not been checked. Take the time limit of the submission of the revised manuscript into account; it is difficult to carry out the first procedure in the revised manuscript. However, we appreciate for the Reviewers’ good advice earnestly. We will check these two procedures in future studies. For the third question proposed by Reviewer, our answer is as following: According to the Reviewer's good instruction, we have recorded Raman spectra of SC in solid state, as shown in Fig. 1(a) (Fig. 4(a) in revised manuscript). Meanwhile, the points 3.2 and 3.3 have also been re-written according to the instructions proposed by the Reviewer. (See the section of 3.2 and 3.3 in the revised manuscript)
图略
Fig1.man spectrum of (a) solid SC. SERS spectra of 1.0×10 mol/L SC aqueous solution adsorbed on (b) glass slide, (c) ******-Ag CNPs, and (d) ******, respectively.
6. Response to comment: Sentence \(page 9) or the number of spectra is 5 (Table 1 and 2), what does it mean? Perhaps something like this: Each
SERS spectrum is recorded 5 times in different points of the ****** surface?
Response: As the Reviewer's good question, the sentence of “the number of spectra for every
condition is five\is hard to be understood. This sentence means that each SERS spectrum is recorded 5 times in differpoints of the ****** surface. we have revised this sentence in the revised manuscript according to the Reviewer's advice.
7. Response to comment: Epigrap****** of Table 1 and 2 should be revised. Corrections are indicated in the manuscript.
Response: According to the Reviewer's good instruction, we have revised the epigrap****** of Table
1 and 2. The epigraph of Table 1 “Table 1 Preliminary assignations of the Raman bands (Mean ± S.D., n=5) for the SERS spectra of ******. The number of spectra is 5” is revised as “Assignments for the SERS bands (Mean ± S.D., n=5) of ****** (based on Refs. [25-30]).”
The epigraph of Table 2 “Table 2 Observed wavenumbers (Mean ± S.D., n=5), assignments, and local coordinates of ******, excited at 785 nm. The number of spectra is 5. [35-37]” is revised as “Table 2 Assignments for the SERS bands (Mean ± S.D., n=5) of ****** (based on Refs. [32-34]).”
8. Response to comment: Figure 4 shows SERS spectra of ****** and ****** recorded on different Ag nanofilms. What do authors want to say? This experiment is to check the reproducibility of the method? Thus, it is better to use the word \\
Response: As the Reviewer's good advice, we should use the word “reproducibility” and not
\****** recorded on the different ******s prepared under the same conditions. The authors want to display the reproducibility of the ******s prepared by this simple method. Thus, we have replaced the
word \
9. Response to comment: Uv-vis absorption spectrum of不******-protected Ag nanoparticules shows a maximum at 418 nm and at 785 nm the absorbance is zero (Figure
1b). Given that SERS spectra are recorded at 785 nm, I think that this excitation line is very far from the maximum to obtain a good signal. In fact, the Raman signal is very weak (Figure 3). Is it possible to obtain a better signal employing another excitation laser, for example, 514nm? I mean, probably the 785nm line is better for ****** than for ******-protected Ag nanoparticules, but under other different excitation laser the ******-protected Ag nanoparticules could be a good substrate for ****** or ******. Have been checked different excitation laser lines?
Response: We are very sorry for our negligence of the explanation for why SERS spectra of ******
and ****** are recorded at 785 nm. In the studying of the SERS effect of ****** and ******, we think that the excitation with the 785 nm wavelength has a number of advantageous features compared to [a] other wavelengt******. A previous study has reported that a laser wavelength shorter than 514.5 nm is known to enhance photodissociation and cause protein degradation even at a low power. However, the sample damage can be avoided using laser light of a longer wavelength. No paling effects were observed using laser light with wavelength ≥660 nm. In their study, degradation of the biological objects was observed when using 514.5 nm excitation lasers. Meanwhile, it is known that when using 660 nm irradiation, for a laser power of 10mW and a diameter of 10 μm, 2the light intensity is up to 127 MW/m . In our system, the laser power is set at 65mW and the 2 3 2 diameter is 90μm. So the light intensity was ca. 10 MW/m (10 W/cm ), which is much smaller than that of 660 nm irradiation. Thus, in our studies, we employed a 65-mW, 785-nm diode laser to record the Raman and SERS spectra of ****** and ****** in order to avoid the photodissociation and degradation of the proteins.
[a] G. J. Puppels, J. H. F. Olminkhof, G. M. J. Segers-Nolten, C. Otto, F. F. M. Demul, J. Greve. Laser Irradiation and Raman Spectroscopy of Single Living Cells and Chromosomes: Sample Degradation Occurs with 514.5 nm but not with 660 nm Laser Light . Exp. Cell Res, 195 (1991) 361.
Special thanks to you for your good comments.
Reviewer 4#
1. Response to comment: I'm not familiar with blood sample preparation but I wasn't able to extract the final concentration of ****** and ****** added to the ******-NPs and ******. This value has to be clearly reported in the text.
Response: As the Reviewer suggested that we have calculated the final concentration of ****** and ****** added to the ******-protected ****** (******-Ag CNPs) and ****** in the revised manuscript. According our calculation, the final concentration of ****** and ****** added to ****** is ca.4.8 and 1.5%, respectively. However, take the dilution of the Ag colloid into account, the final concentration of ****** and ****** added to the ******-Ag CNPs is ca.0.27 and 0.08 %, respectively. All these concentrations have been added in the revised manuscript.Although the final concentrations of ****** added to the ******-NPs and ****** are different, the quantity of ****** lighted by the laser