Raman microspectroscopy has become an effective method for analyzing the molecular appearance of biomarkers in skin tissue. For the first time, we acquired in vitro Raman spectra of healthy and malignant skin tissues, including basal cell carcinoma (BCC) and squamous cell carcinoma (SCC), at 532 and 785 nm laser excitation wavelengths in the wavenumber ranges of 900–1800 cm−1 and 2800–3100 cm−1 and analyzed them to find spectral features for differentiation between the three classes of the samples. The intensity ratios of the bands at 1268, 1336, and 1445 cm−1 appeared to be the most reliable criteria for the three-class differentiation at 532 nm excitation, whereas the bands from the higher wavenumber region (2850, 2880, and 2930 cm−1) were a robust measure of the increased protein/lipid ratio in the tumors at both excitation wavelengths. Selecting ratios of the three bands from the merged (532 + 785) dataset made it possible to increase the accuracy to 87% for the three classes and reach the specificities for BCC + SCC equal to 87% and 81% for the sensitivities of 95% and 99%, respectively. Development of multi-wavelength excitation Raman spectroscopic techniques provides a versatile non-invasive tool for research of the processes in malignant skin tumors, as well as other forms of cancer.
In this paper, we report the study of the low-voltage locally enhanced electric field treatment (LEEFT) of E. coli bacteria via TEM analysis and FT-IR and Raman spectrometry. The formation of pores was confirmed by TEM, which revealed the membrane rupture along with the formation of precipitates in the membrane and the cell volume, and by photoluminescence spectra of propidium iodide dye. LEEFT results in the alternation of DNA and RNA structure, indicated by the change in its α-helical and β-helical forms, decrease and narrowing of the nucleic acids-related IR and Raman peaks. Membrane damage is illustrated by the ambiguous character after low-voltage exposure: several membrane fatty acids’ peaks are broadening, while the others narrow, which indicates the overall change of the molecular bonds in their moiety, and may result from the rigidification during the shrinkage of the inner membrane and the increase in fluidity of the outer membrane. The corresponding fingerprints of cytochrome c and bo, located in the membrane or in the periplasm, on Raman spectra, reflect the arising changes in their structure and moiety. The observed changes were partially confirmed by TEM images, which indicate the dense precipitates’ formation in the cell volume and membrane, as well as the cytoplasmic membrane shrinkage away from the outer membrane.
We report the successful inactivation of P. aeruginosa strain by femtosecond infrared (IR) laser radiation at the resonant wavelengths of 3.15 μm and 6.04 μm, chosen due to the presence of characteristic molecular vibrations in the main structural elements of the bacterial cells in these spectral ranges: vibrations of amide groups in proteins (1500–1700 cm−1), and C-H vibrations in membrane proteins and lipids (2800–3000 cm−1). The underlying bactericidal structural molecular changes were revealed by the stationary Fourier-transform IR spectroscopy, with the spectral peaks parameters being obtained by Lorentzian fitting with the hidden peaks revealed by the second derivative calculations, while no visible damage to the cell membranes was identified by scanning and transmission electron microscopy.
We report the bactericidal effect of Ag and Cu NPs with different concentrations on methicillin-resistant S. aureus strain in comparison to the effect of AgNO3 and CuCl2 solutions, characterized by microbiological tests, TEM and Fourier-transform infrared spectroscopy. NPs were produced by nanosecond laser ablation in distilled water and characterized by scanning electron microscopy, UV-vis, energy dispersive X-ray, FT-IR spectroscopy, as well as X-ray diffraction, dynamic light scattering size and zeta-potential measurements. Microbiological tests showed antibacterial activity of NPs and metal ion-containing salts. Comparative FT-IR spectroscopy of bacteria, treated with metal NPs and salts, showed the broadening of amide I and II bands, a CH2-related peak and its frequency decrease, indicating the increase of membrane fluidity. The main mechanisms of the antibacterial effect were proposed: Ag and Cu NPs release ions and ROS, which result in lipid peroxidation; AgNO3 forms precipitates on the cell surface, which lead to the mechanical rupture of the membrane and subsequent possible penetration of the precipitates in the emerged damaged spots, complete destruction of the membrane and bacterial death; Cu ions from the CuCl2 solution cause damage to phosphorus- and sulfur-containing biomolecules, which leads to disruption of intracellular biochemical processes. The theories were confirmed by FT-IR spectroscopy and TEM.
Confocal scanning Raman and photoluminescence (PL) microspectroscopy is a structure-sensitive optical method that allows the non-invasive analysis of biomarkers in the skin tissue. We used it to perform in vitro diagnostics of different malignant skin neoplasms at several excitation wavelengths (532, 785 and 1064 nm). Distinct spectral differences were noticed in the Raman spectra of basal cell carcinoma (BCC) and squamous cell carcinoma (SCC), compared with healthy skin. Our analysis of Raman/PL spectra at the different excitation wavelengths enabled us to propose two novel wavelength-independent spectral criteria (intensity ratios for 1302 cm−1 and 1445 cm−1 bands, 1745 cm−1 and 1445 cm−1 bands), related to the different vibrational “fingerprints” of cell membrane lipids as biomarkers, which was confirmed by the multivariate curve resolution (MCR) technique. These criteria allowed us to differentiate healthy skin from BCC and SCC with sensitivity and specificity higher than 95%, demonstrating high clinical importance in the differential diagnostics of skin tumors.
The single-pass one-step method for printing conductive silver tracks on a glass surface, using the laser-induced forward transfer (LIFT) technique, was proposed, providing a unique opportunity for high-throughput printing of surface micro- and nanostructures with high electrical conductivity and positioning accuracy. This method was developed via our multi-parametric research, resulting in the selection of the optimal material, laser irradiation, and transfer conditions. Optical, scanning and transmission electron, and atomic force microscopy methods, as well as X-ray diffraction, were used to characterize the surface structure and phase state of the printed structures, while energy-dispersive X-ray and X-ray photoelectron microscopy were employed for their chemical microanalysis. Depending on the laser irradiation parameters, the specific electrical conductivity of the printed tracks varied from 0.18 to 83 kS/cm, approaching that of donor magnetron-sputtered films. This single-pass one-step method significantly facilitates fast, large-scale, on-demand local laser printing of metallic (sub)microcomponents of microelectronic devices.