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      Estimation of the age of human semen stains by attenuated total reflection Fourier transform infrared spectroscopy: a preliminary study

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          Abstract

          Semen stain is one of the most important biological evidence at sexual crime scenes. Age estimation of human semen stains plays an important role in forensic work, and it is rarely studied due to lack of well-established methods. In this study, the technique called attenuated total reflection Fourier transform infrared spectroscopy (ATR-FTIR) coupled with advanced chemometric methods was employed to determine the age of semen stains on three different substrates: glass slides, tissues and fabric made of regenerated cellulose fibres up to 6 d. Partial least squares regression (PLSR) was used in conjunction with spectral analysis for age estimation, and the results generated high R 2 values (cross-validation: 0.81, external validation: 0.74) but a narrow margin of error for root mean square error (RMSE) (RMSE of cross-validation: 0.77 d, RMSE of prediction: 1.02 d). Additionally, our results indicated the robustness of PLSR model was not weaken by the influence of different substrates in this study. Our results indicate that ATR-FTIR, combined with chemometric methods, shows great potential as a convenient and efficient tool for age estimation of semen stains. Moreover, the method could be applied to routine forensic investigations in the future.

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          Using Fourier transform IR spectroscopy to analyze biological materials.

          IR spectroscopy is an excellent method for biological analyses. It enables the nonperturbative, label-free extraction of biochemical information and images toward diagnosis and the assessment of cell functionality. Although not strictly microscopy in the conventional sense, it allows the construction of images of tissue or cell architecture by the passing of spectral data through a variety of computational algorithms. Because such images are constructed from fingerprint spectra, the notion is that they can be an objective reflection of the underlying health status of the analyzed sample. One of the major difficulties in the field has been determining a consensus on spectral pre-processing and data analysis. This manuscript brings together as coauthors some of the leaders in this field to allow the standardization of methods and procedures for adapting a multistage approach to a methodology that can be applied to a variety of cell biological questions or used within a clinical setting for disease screening or diagnosis. We describe a protocol for collecting IR spectra and images from biological samples (e.g., fixed cytology and tissue sections, live cells or biofluids) that assesses the instrumental options available, appropriate sample preparation, different sampling modes as well as important advances in spectral data acquisition. After acquisition, data processing consists of a sequence of steps including quality control, spectral pre-processing, feature extraction and classification of the supervised or unsupervised type. A typical experiment can be completed and analyzed within hours. Example results are presented on the use of IR spectra combined with multivariate data processing.
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            Distinguishing cell types or populations based on the computational analysis of their infrared spectra.

            Infrared (IR) spectroscopy of intact cells results in a fingerprint of their biochemistry in the form of an IR spectrum; this has given rise to the new field of biospectroscopy. This protocol describes sample preparation (a tissue section or cytology specimen), the application of IR spectroscopy tools, and computational analysis. Experimental considerations include optimization of specimen preparation, objective acquisition of a sufficient number of spectra, linking of the derived spectra with tissue architecture or cell type, and computational analysis. The preparation of multiple specimens (up to 50) takes 8 h; the interrogation of a tissue section can take up to 6 h (∼100 spectra); and cytology analysis (n = 50, 10 spectra per specimen) takes 14 h. IR spectroscopy generates complex data sets and analyses are best when initially based on a multivariate approach (principal component analysis with or without linear discriminant analysis). This results in the identification of class clustering as well as class-specific chemical entities.
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              The detection and discrimination of human body fluids using ATR FT-IR spectroscopy.

              Blood, saliva, semen and vaginal secretions are the main human body fluids encountered at crime scenes. Currently presumptive tests are routinely utilised to indicate the presence of body fluids, although these are often subject to false positives and limited to particular body fluids. Over the last decade more sensitive and specific body fluid identification methods have been explored, such as mRNA analysis and proteomics, although these are not yet appropriate for routine application. This research investigated the application of ATR FT-IR spectroscopy for the detection and discrimination of human blood, saliva, semen and vaginal secretions. The results demonstrated that ATR FT-IR spectroscopy can detect and distinguish between these body fluids based on the unique spectral pattern, combination of peaks and peak frequencies corresponding to the macromolecule groups common within biological material. Comparisons with known abundant proteins relevant to each body fluid were also analysed to enable specific peaks to be attributed to the relevant protein components, which further reinforced the discrimination and identification of each body fluid. Overall, this preliminary research has demonstrated the potential for ATR FT-IR spectroscopy to be utilised in the routine confirmatory screening of biological evidence due to its quick and robust application within forensic science.
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                Author and article information

                Journal
                Forensic Sci Res
                Forensic Sci Res
                TFSR
                tfsr20
                Forensic Sciences Research
                Taylor & Francis
                2096-1790
                2471-1411
                2020
                09 September 2019
                : 5
                : 2
                : 119-125
                Affiliations
                [a ]Department of Forensic Pathology, Xi'an Jiaotong University School of Medicine , Xi’an, China;
                [b ]Department of Forensic Medicine, Chongqing Medical University , Chongqing, China;
                [c ]Department of Forensic Medicine, Xuzhou Medical University , Xuzhou, China;
                [d ]Department of Forensic Pathology, Academy of Forensic Science , Shanghai, China
                Author notes
                CONTACT Ping Huang huangp@ 123456ssfd.cn ;
                Author information
                http://orcid.org/0000-0002-4189-9580
                Article
                1642567
                10.1080/20961790.2019.1642567
                7476623
                5a1c59b0-db1d-4e71-8ff7-b9b354e606e3
                © 2019 The Author(s). Published by Taylor & Francis Group on behalf of the Academy of Forensic Science.

                This is an Open Access article distributed under the terms of the Creative Commons Attribution License ( http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

                History
                : 07 June 2018
                : 04 July 2019
                : 09 July 2019
                Page count
                Figures: 7, Tables: 2, Pages: 7, Words: 4408
                Funding
                Funded by: National Natural Science Foundation of China 10.13039/501100001809
                This work was supported by the National Natural Science Foundation of China (grant number 81730056).
                This work was supported by the National Natural Science Foundation of China [grant number 81730056].
                Categories
                Original Articles

                forensic sciences,forensic medicine,semen stain,age estimation,spectroscopy,fourier transform infrared,chemometrics

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