Forensic analysis of biological fluid stains on substrates by spectroscopic approaches and chemometrics: A review.

BACKGROUND: Bodily fluid stains are one of the most relevant evidence that can be found at the crime scene as it provides a wealth of information to the investigators. They help to report on the individuals involved in the crime, to check alibis, or to determine the type of crime that has been committed. They appear as stains in different types of substrates, some of them porous, which can interfere in the analysis. The spectroscopy techniques combined with chemometrics are showing increasing potential for their use in the analysis of such samples due to them being fast, sensitive, and non-destructive. FINDINGS: This is a comprehensive review of the studies that used different spectroscopic techniques followed by chemometrics for analysing biological fluid stains on several surfaces, and under various conditions. It focuses on the bodily fluid stains and the most suitable spectroscopic techniques to study forensic scientific problems such as the substrate's characteristics, the influence of ambient conditions, the aging process of the bodily fluids, the presence of animal bodily fluids and non-biological fluids (interfering substances), and the bodily fluid mixtures. The most widely used techniques were Raman spectroscopy and attenuated total reflection Fourier transform infrared spectroscopy (ATR FTIR). Nonetheless, other non-destructive techniques have been also used, like near infrared hyperspectral imaging (HSI-NIR) or surface enhanced Raman spectroscopy (SERS), among others. This work provides the criteria for the selection of the most promising non-destructive techniques for the effective in situ detection of biological fluid stains at crime scene investigations. SIGNIFICANCE AND NOVELTY: The use of the proper spectroscopic and chemometric approaches on the crime scene is expected to improve the support of forensic sciences to criminal investigations. Evidence may be analysed in a non-destructive manner and kept intact for further analysis. They will also speed up forensic investigations by allowing the selection of relevant samples from occupational ones.

Crime Light Imaging (CLI): A Novel Sensor for Stand-Off Detection and Localization of Forensic Traces.

Stand-off detection of latent traces avoids the scene alteration that might occur during close inspection by handheld forensic lights. Here, we describe a novel sensor, named Crime Light Imaging (CLI), designed to perform high-resolution photography of targets at a distance of 2-10 m and to visualize some common latent traces. CLI is based on four high-power illumination LEDs and one color CMOS camera with a motorized objective plus frontal filters; the LEDs and camera could be synchronized to obtain short-exposure images weakly dependent on the ambient light. The sensor is integrated into a motorized platform, providing the target scanning and necessary information for 3D scene reconstruction. The whole system is portable and equipped with a user-friendly interface. The preliminary tests of CLI on fingerprints at distance of 7 m showed an excellent image resolution and drastic contrast enhancement under green LED light. At the same distance, a small (1 µL) blood droplet on black tissue was captured by CLI under NIR LED, while a trace from 15 µL semen on white cotton became visible under UV LED illumination. These results represent the first demonstration of true stand-off photography of latent traces, thus opening the way for a completely new approach in crime scene forensic examination.

Differentiation of blood and environmental interfering stains on substrates by Chemometrics-Assisted ATR FTIR spectroscopy.

Blood is the most common and relevant bodily fluid that can be found in crime scenes. It is critical to correctly identify it, and to be able to differentiate it from other substances that may also appear at the crime scene. In this work, several stains of blood, chocolate, ketchup, and tomato sauce on five different substrates (plywood, metal, gauze, denim, and glass) were analysed by ATR FTIR spectroscopy assisted with orthogonal partial least square-discriminant analysis (OPLS-DA) models. It was possible to differentiate blood from the environmental interfering substances independently of the substrate they were on, and to differentiate bloodstains according to the substrate they were deposited on. These results represent a proof-of-concept that open new horizons to differentiate bloodstains from other interfering substances on common substrates present in crime scenes.