Light and Autofluorescence, Multitasking Features in Living Organisms
Round 1
Reviewer 1 Report
Croce presented an extensive review of endogenous molecules and their relevance in research existing in plants, animals, and mammals. The review provides an excellent overview of these autofluorescence species. The manuscript is well written. I have a few minor comments.
- A chart showing the chronological event/discoveries mentioned in section 1 would make it clearer.
- Figure 1, it will be helpful for readers to have the excitation and emission wavelength values for the vegetable fluorescent compounds highlighted here perhaps as a table.
- Could the author include a table of fluorophores discussed, their spectral properties, and research application for each section?
- Some of the texts in Figure 15 are hard to read. Could the font size be increased? Also, it will be helpful to the reader if a more detailed description of the figure was included in the caption.
- Text in Figure 21 is hard to read.
- Sections 6, flavins like FAD and FMN should be discussed. Also, define optical redox ratio (line 1265) and its implications.
- The manuscript should include some description/description of lifetime imaging (1285) else it appears vague.
Author Response
I thank very much the Reviewer for the time spent in reading the paper and for the kind remarks, surely helping to improve and make the text clearer to the readers. All remarks have been carefully considered and the paper has been revised accordingly, as reported point by point below. I wish that this will help to improve the interest of the scientific community in the potential of fluorescence / autofluorescence applications in biology, making easier to have an overlook of the various related arguments also to colleague working in a specific fluorescence field.
Point by point answers to the Reviewer’s remarks:
Q- A chart showing the chronological event/discoveries mentioned in section 1 would make it clearer.
A – Many thanks for the suggestion, surely valuable in helping to make clearer the time course of some main steps in the history of autofluorescence and its applications. The chart has been added to the text, as the new Figure 1. Subsequent figures have been renumbered accordingly.
Q -Figure 1, it will be helpful for readers to have the excitation and emission wavelength values for the vegetable fluorescent compounds highlighted here perhaps as a table.
A- A table (Table 1) has been added to the text, reporting the exc/em wavelengths of the mostly vegetable fluorescing fluorophores recalled in Figure 1 (now Figure 2).
Q - Could the author include a table of fluorophores discussed, their spectral properties, and research application for each section?
A- Tables have been added to the text, one for each section, reporting the main fluorophores of the section, along with indicative exc / em values, applications, and references. Tables have resulted in the addition of some new references. Therefore, references have been reorganized along the overall text.
Q - Some of the texts in Figure 15 are hard to read. Could the font size be increased? Also, it will be helpful to the reader if a more detailed description of the figure was included in the caption.
A – I agree that reading is not easy. The figure is reproduced from a published paper, and a new effort has been made to improve the reading of the characters. Also, the description in the caption has been implemented.
Q - Text in Figure 21 is hard to read.
A - Also in this case, being the figure reproduced from a published paper, an effort has been made to improve the reading of the characters, by enlarging the two parts of the figure, now split and presented side by side.
Q - Sections 6, flavins like FAD and FMN should be discussed. Also, define optical redox ratio (line 1265) and its implications.
A – I agree with the Reviewer that flavins needed a better description, as well as redox ratio. Section 6 has been revised accordingly; related text is given in red (lines 1314-1341).
Q- The manuscript should include some description/description of lifetime imaging (1285) else it appears vague.
A – Also here I agree with the Reviewer that lifetime imaging needed to be better defined. The revised text is given in red (lines 1350-1359).
Reviewer 2 Report
The presented paper is an impressive review of the field of biofluorescence. To the best of my knowledge, such a wide range of fluorophores and systems has never been systematically analyzed within a single paper. I'm sure that this review would make a great contribution to the field, providing a broader outlook to the origin of fluorophores to researchers from different areas, thus extending their knowledgebase. Indeed, as the author writes in the abstract, the papers on biofluorescence are endless, and some subjects are ommitted, but this fact would hold true for any review of similar broadness. Summarizing, I think that the paper should be accepted for publication in Photochem as it is.
Author Response
I thank very much the Reviewer for the time spent in considering my paper and for the kind appreciation. I really wish it will help to increase the interest in this branch of photobiology from the scientific community, promoting an overlook of the various arguments also to colleague working in specific fluorescence/ autofluorescence areas.
