different approaches to ft ir microspectroscopy on x ray exposed human cells

Fourier-Transform Infrared microspectroscopy (μFT-IR) has been usefully applied in the analysis of the complex biological processes occurring during X-ray radiation-cell interaction. Different experimental approaches are available for FT-IR spectra collection (transmission, attenuated total reflection (ATR), and transflection modes) from cells samples. Recently, some problems have been raised about the role of transmitted and reflected components of the infrared beam in transflection mode. For this reason, we investigated two different transflection approaches for collecting spectra from cells exposed to X-ray. In the former approach, cells were grown on MirrIR slides, and for the second approach, cell pellets were prepared. In both cases, SH-SY5Y neuroblastoma cells were used. X-ray exposure was performed at doses of 2 and 4 Gy. Spectra were obtained by using both the approaches in the 600–4000 cm−1 spectral range from exposed and not-exposed samples. The main contributions from proteins, lipids, carbohydrates, and DNA were clearly evidenced in spectra obtained with the two different acquisition approaches. A comparison among them has been also reported

laser safety standards and measurements of hazard parameters for medical lasers

Laser sources are nowadays largely adopted in medicine and hence they are widespread in medical environment, where patients are present and the users are not always highly specialized in managing laser sources. This has greatly boosted the attention towards safety issues related to exposure to laser beams and to strictly assess the values of well defined laser radiation standard parameters characterizing the level of hazard of laser sources. In this framework, we measured two of the most important parameters, Maximum Permissible Exposure (MPE) and Nominal Ocular Hazard Distance (NOHD), for some of the laser sources mostly employed in medicine. Additionally, we compared our results with data elaborated from standards in order to single out safe and comfortable working conditions. The results here reported have shown that information provided by manufacturers is often not enough to define the hazard level of the laser source and measurements of the main safety parameters are mandatory

preparation and characterization of au nanoparticles for teranostic applications

Gold nanoparticles (GNPs) are very attractive materials due to their unique properties of small size, large surface area to volume ratio, high reactivity to the living cells, stability over high temperatures. These properties along with the evidence that GNPs are amenable to the attachment of biomolecules or ligands through well-known thiol and amino chemistry or simply by electrostatic interactions have led to a wealth of nanoparticle-based bio-devices for many teranostic applications and the research effort in the field is still huge. In the present work absorption spectroscopy, static and dynamic light scattering, Fourier-Transform infrared (FT-IR) microspectroscopy and TEM microscopy have been used to characterize different sized bare and biotinylated GNPs (from 20 to 70 nm diameter). These experimental techniques have been also applied to investigate the aggregation process of the biotinylated particles induced by addition of neutravidin at various concentrations in the nanomolar range. In particular, optical visible techniques have been used for estimating the size of particles before and after the biotin capping procedure. FT-IR microspectroscopy has allowed us to investigate the biochemical changes occurring in GNPs after interaction processes while TEM microscopy has enabled to observe the relative morphological modifications. Moreover, the outlined modifications in the scattering properties have been related to changes in the size also thanks to numerical evaluation of scattering cross section by using Mie theory. The complete characterization of these processes is of fundamental importance for further manipulations required for GNPs teranostic applications

Application of Vibrational Spectroscopies in the Qualitative Analysis of Gingival Crevicular Fluid and Periodontal Ligament during Orthodontic Tooth Movement

Optical vibrational techniques show a high potentiality in many biomedical fields for their characteristics of high sensitivity in revealing detailed information on composition, structure, and molecular interaction with reduced analysis time. In the last years, we have used these techniques for investigating gingival crevicular fluid (GCF) and periodontal ligament (PDL) during orthodontic tooth treatment. The analysis with Raman and infrared signals of GCF and PDL samples highlighted that different days of orthodontic force application causes modifications in the molecular secondary structure at specific wavenumbers related to the Amide I, Amide III, CH deformation, and CH3/CH2. In the present review, we report the most relevant results and a brief description of the experimental techniques and data analysis procedure in order to evidence that the vibrational spectroscopies could be a potential useful tool for an immediate monitoring of the individual patient's response to the orthodontic tooth movement, aiming to more personalized treatment reducing any side effects

FT-IR Transflection Micro-Spectroscopy Study on Normal Human Breast Cells after Exposure to a Proton Beam

Fourier transform infrared micro-spectroscopy (mu-FT-IR) is nowadays considered a valuable tool for investigating the changes occurring in human cells after exposure to ionizing radiation. Recently, considerable attention has been devoted to the use of this optical technique in the study of cells exposed to proton beams, that are being increasingly adopted in cancer therapy. Different experimental configurations are used for proton irradiation and subsequent spectra acquisition. To facilitate the use of mu-FT-IR, it may be useful to investigate new experimental approaches capable of speeding up and simplifying the irradiation and measurements phases. Here, we propose the use of low-e-substrates slides for cell culture, allowing the irradiation and spectra acquisition in transflection mode in a fast and direct way. In recent years, there has been a wide debate about the validity of these supports, but many researchers agree that the artifacts due to the presence of the electromagnetic standing wave effects are negligible in many practical cases. We investigated human normal breast cells (MCF-10 cell line) fixed immediately after the irradiation with graded proton radiation doses (0, 0.5, 2, and 4 Gy). The spectra obtained in transflection geometry showed characteristics very similar to those present in the spectra acquired in transmission geometry and confirm the validity of the chosen approach. The analysis of spectra indicates the occurrence of significant changes in DNA and lipids components of cells. Modifications in protein secondary structure are also evidenced

Micro-Raman Spectroscopy and Univariate Analysis for Monitoring Disease Follow-Up

Micro-Raman spectroscopy is a very promising tool for medical applications, thanks to its sensitivity to subtle changes in the chemical and structural characteristics of biological specimens. To fully exploit these promises, building a method of data analysis properly suited for the case under study is crucial. Here, a linear or univariate approach using a R2 determination coefficient is proposed for discriminating Raman spectra even with small differences. The validity of the proposed approach has been tested using Raman spectra of high purity glucose solutions collected in the 600 to 1,600 cm−1 region and also from solutions with two known solutes at different concentrations. After this validation step, the proposed analysis has been applied to Raman spectra from oral human tissues affected by Pemphigus Vulgaris (PV), a rare life-threatening autoimmune disease, for monitoring disease follow-up. Raman spectra have been obtained in the wavenumber regions from 1,050 to 1,700 cm−1 and 2,700 to 3,200 cm−1 from tissues of patients at different stages of pathology (active PV, under therapy and PV in remission stage) as confirmed by histopathological and immunofluorescence analysis. Differences in the spectra depending on tissue illness stage have been detected at 1,150–1,250 cm−1 (amide III) and 1,420–1,450 cm−1 (CH3 deformation) regions and around 1,650 cm−1 (amide I) and 2,930 cm−1 (CH3 symmetric stretch). The analysis of tissue Raman spectra by the proposed univariate method has allowed us to effectively differentiate tissues at different stages of pathology