O the reaction of O2(1Dg) with TPP triplets inside the polymeric nanofibers (Fig. 2B) [30]. The advantage of this technique compared to direct detection of O2(1Dg) via phosphorescence is its higher signal-to-noise ratio; Calciferol web however, the kinetics of SODF are complicated and do not allow estimation of lifetimes (tT and tD) through a simple fitting process. Fluorescence lifetime imaging microscopy made it possible to distinguish between the immediate fluorescence that arises from TPP when it is directly excited and the light from SODF, which is Verubecestat dependent on the concentrations of O2(1Dg) and TPP triplets (Figure 3b) [31]. While the immediate fluorescence intensity image (Figure 3A) shows the distribution of TPP molecules inside nanofibers, the SODF intensity image reveals domains with different concentrations of O2(1Dg) (Fig. 3B). It should be notedthat the diffraction-limited spatial 23727046 resolution of both images is approximately 200 nm. The method of O2(1Dg) imaging using SODF does not monitor O2(1Dg) outside of the nanofibers. It should be noted that the average diameters of the nanofibers (ca 90 nm for Tecophilic and ca 200 nm for PCL) are sufficiently small for O2(1Dg) to effectively diffuse outside of the nanofibers and directly interact with viruses. The average diffusion length of O2(1Dg) depends on the diffusion coefficient in the polymer; a typical value is several tens to hundredths of nm for tD within a range of 10 to 25 ms [31]. Although tD in the surrounding aqueous media falls to 3.1 ms [32], the diffusion length remains unchanged or increases because the diffusion coefficient of oxygen in water is one or two orders of magnitude higher than that in a polymer.Figure 3. Distribution of TPP molecules in the nanofibers. Confocal fluorescence microscopy: fluorescence intensity images (20620 mm) of TPP in the TecophilicH nanofiber textile based on the data collected 10?0 ns after excitation (prompt fluorescence) (a) and 300?000 ns after excitation (SODF) (b). doi:10.1371/journal.pone.0049226.gVirucidal Nanofiber TextilesPhotooxidation of 9,10-anthracenediylbis(methylene)dimalonic acid (AMA) on the surface of the nanofiber textiles doped with TPPThe results from both luminescence spectroscopy and microscopy described above presented clear evidence of O2(1Dg) photogeneration inside the polymeric nanofibers. We next asked whether O2(1Dg) could diffuse from the nanofibers to the textile surface and oxidize a substrate. As a suitable substrate, we selected AMA, a known water-soluble singlet oxygen trap [33]. Continuous visible light irradiation (see Materials and Methods) of a piece of the nanofiber textile immersed in a detection solution of AMA in air-saturated water resulted in significant spectral changes, indicating photooxidation of AMA to corresponding endoperoxides (Fig. 4). No spectral changes were observed in the absence of light or oxygen (the detection solution was bubbled with N2). Furthermore, irradiation of the nanofiber textile without TPP photosensitizer did not induce any AMA photoxidation.Photovirucidal effect of the nanofiber textiles doped with TPPWe next asked whether the O2(1Dg) released from the surface of the TPP-doped hydrophilic nanofiber textiles could inactivate viruses. Therefore, we examined the effect of O2(1Dg) released from the textiles on viruses falling into two different categories: polyomaviruses, the genomes of which are protected by a proteinaceous coat composed of viral capsid proteins (nonenveloped vir.O the reaction of O2(1Dg) with TPP triplets inside the polymeric nanofibers (Fig. 2B) [30]. The advantage of this technique compared to direct detection of O2(1Dg) via phosphorescence is its higher signal-to-noise ratio; however, the kinetics of SODF are complicated and do not allow estimation of lifetimes (tT and tD) through a simple fitting process. Fluorescence lifetime imaging microscopy made it possible to distinguish between the immediate fluorescence that arises from TPP when it is directly excited and the light from SODF, which is dependent on the concentrations of O2(1Dg) and TPP triplets (Figure 3b) [31]. While the immediate fluorescence intensity image (Figure 3A) shows the distribution of TPP molecules inside nanofibers, the SODF intensity image reveals domains with different concentrations of O2(1Dg) (Fig. 3B). It should be notedthat the diffraction-limited spatial 23727046 resolution of both images is approximately 200 nm. The method of O2(1Dg) imaging using SODF does not monitor O2(1Dg) outside of the nanofibers. It should be noted that the average diameters of the nanofibers (ca 90 nm for Tecophilic and ca 200 nm for PCL) are sufficiently small for O2(1Dg) to effectively diffuse outside of the nanofibers and directly interact with viruses. The average diffusion length of O2(1Dg) depends on the diffusion coefficient in the polymer; a typical value is several tens to hundredths of nm for tD within a range of 10 to 25 ms [31]. Although tD in the surrounding aqueous media falls to 3.1 ms [32], the diffusion length remains unchanged or increases because the diffusion coefficient of oxygen in water is one or two orders of magnitude higher than that in a polymer.Figure 3. Distribution of TPP molecules in the nanofibers. Confocal fluorescence microscopy: fluorescence intensity images (20620 mm) of TPP in the TecophilicH nanofiber textile based on the data collected 10?0 ns after excitation (prompt fluorescence) (a) and 300?000 ns after excitation (SODF) (b). doi:10.1371/journal.pone.0049226.gVirucidal Nanofiber TextilesPhotooxidation of 9,10-anthracenediylbis(methylene)dimalonic acid (AMA) on the surface of the nanofiber textiles doped with TPPThe results from both luminescence spectroscopy and microscopy described above presented clear evidence of O2(1Dg) photogeneration inside the polymeric nanofibers. We next asked whether O2(1Dg) could diffuse from the nanofibers to the textile surface and oxidize a substrate. As a suitable substrate, we selected AMA, a known water-soluble singlet oxygen trap [33]. Continuous visible light irradiation (see Materials and Methods) of a piece of the nanofiber textile immersed in a detection solution of AMA in air-saturated water resulted in significant spectral changes, indicating photooxidation of AMA to corresponding endoperoxides (Fig. 4). No spectral changes were observed in the absence of light or oxygen (the detection solution was bubbled with N2). Furthermore, irradiation of the nanofiber textile without TPP photosensitizer did not induce any AMA photoxidation.Photovirucidal effect of the nanofiber textiles doped with TPPWe next asked whether the O2(1Dg) released from the surface of the TPP-doped hydrophilic nanofiber textiles could inactivate viruses. Therefore, we examined the effect of O2(1Dg) released from the textiles on viruses falling into two different categories: polyomaviruses, the genomes of which are protected by a proteinaceous coat composed of viral capsid proteins (nonenveloped vir.
