# Soft X-ray Microscopy Techniques for Medical and Biological Imaging at TwinMic--[PERSON]
## 4 Discussion and Conclusions  
In the present paper, the TwinMic imaging and spectroscopic potentialities in biology and biomedicine were highlighted by showing relevant examples in the life sciences field. The possibility of deploying different imaging modalities coupled with X-ray fluorescence spectroscopy makes TwinMic a very attractive tool for the life sciences community.  
Although TwinMic has been successfully employed to provide answers or information for different research topics, there is a continuous trend to improve its capabilities in terms of: (i) sensitivity, by means of new XRF detector systems and new imaging detectors, such as CMOS ones; (ii) spatial resolution, by means of more reliable sample stages, higher-resolution optics systems, and last but not least, by improving the ptychography imaging mode; (iii) chemical element accessibility, by extending its energy range until 4.2 keV, as foreseen in the next years to come.  
TwinMic upgrades will also be pushed forward by new possible requirements or wishes from the life sciences community.  
Another successful key point of TwinMic, which has not been stressed in the current paper, is that its setup modularity and flexibility (i.e., motors stages and sample holders) allow it to be easily complemented with other synchrotron techniques, such as FTIR spectromroscopy [42], XRF at higher energies [37; 38], and microtomography [34], often by analyzing the very same specimens.  
The following are available online at [[https://www.mdpi.com/article/10.3390/app11167216/s1](https://www.mdpi.com/article/10.3390/app11167216/s1)]([https://www.mdpi.com/article/10.3390/app11167216/s1](https://www.mdpi.com/article/10.3390/app11167216/s1)), Figure S1: (A) Ptychographic reconstruction of a 20 \(\upmu\)m diameter tungsten Siemens Star test pattern. (B) Zoom of the area indicated with the red box in Panel A. (C) Profile graph extracted along the red line indicated in Panel B, showing an achievable spatial resolution better than 50 nm.  
Conceptualization, A.G. and G.K.; methodology, A.G., F.B. and G.K.; software, R.B., F.B. and G.K.; formal analysis, V.B., G.G., F.G., F.B. and G.K.; investigation, A.G. and L.P.; resources, A.G. and L.P.; data curation, F.B. and G.K.; writing--original draft preparation, A.G.; writing--review and editing, A.G., V.B., G.G., L.P., F.B. and G.K.; visualization, A.G., V.B., G.G. and F.G. All authors have read and agreed to the published version of the manuscript.  
This research received no external funding. This research acknowledges the support of the internal Elettra Sincrotrone Trieste project Advanced Integrated Imaging Initiative.  
For the human ovarian tissue and human sperm samples, the ethical committees' statements are the ones already produced for the already published works [36; 78; 85]. No new samples were analyzed in this study. For the coronary artery, all procedures were carried out according to international guidelines of animal protection and were approved by the Ethics Committee on Animal Experimentation of the State University of Rio de Janeiro (Process CEA/018/2018), as done for [81], as no new samples were analyzed.  
Informed Consent Statement: Informed consent was obtained from all subjects involved in the studies.  
The data presented in this study are available on request from the corresponding author. The data are not publicly available due to privacy reasons including an embargo period according to the facility's scientific data policy.  
The authors are grateful to [PERSON] from JRC (Ispra, Italy) for providing the fibroblast cells exposed to CoFe\({}_{2}\)O\({}_{4}\) NPs; to [PERSON], [PERSON], and [PERSON] from the University of Rio de Janeiro (Brazil) for the insect and coronary artery samples.  
The authors declare no conflict of interest.