Investigation into the merits of infrared imaging in the investigation of tattoos postmortem

Infrared imaging has a history of use in the forensic examination of artwork and documents and is investigated here for its wider use in the detection of tattoos on the human body postmortem. Infrared photographic and reflectographic techniques were tested on 18 living individuals, displaying a total of 30 tattoos. It was observed that neither age, sex, age of the tattoo, nor, most significantly, skin color affected the ability to image the tattoos using infrared imaging techniques. Second, a piglet carcass was tattooed and the impact of the decomposition process on the visibility of the tattoos assessed. Changes were recorded for 17 days and decomposition included partial mummification and skin discoloration. Crucially, the discoloration was recorded as greatly affecting the image quality using conventional photography, but was insignificant to the infrared recording of these tattoos. It was concluded that infrared reflectography was beneficial in the investigation into tattoos postmortem.     

Computer‐aided Dental Identification: An Objective Method for Assessment of Radiographic Image Similarity*

Abstract: A pilot study evaluated a computer‐based method for comparing digital dental images, utilizing a registration algorithm to correct for variations in projection geometry between images prior to a subtraction analysis. A numerical assessment of similarity was generated for pairs of images. Using well‐controlled laboratory settings, the method was evaluated as to its ability to identify the correct specimen with positive results. A subsequent clinical study examined longitudinal radiographic examinations of selected anatomical areas on 47 patients, analyzing the computer‐based method in making the correct identification based upon a threshold level of similarity. The results showed that at a threshold of 0.855, there were two false negative and two false positive identifications out of 957 analyses. Based on these initial findings, 25 dental records having two sets of full mouth series of radiographs were selected. The radiographs were digitized and grouped into six anatomical regions. The more recent set of films served as postmortem images. Each postmortem image was analyzed against all other images within the region. Images were registered to correct for differences in projection geometry prior to analysis. An area of interest was selected to assess image similarity. Analysis of variance was used to determine that there was a significant difference between images from the same individual and those from different individuals. Results showed that the threshold level of concordance will vary with the anatomical region of the mouth examined. This method may provide the most objective and reliable method for postmortem dental identification using intra‐oral images.     

Identification of the Edentulous Individual: An Investigation into the Accuracy of Radiographic Identifications*

Abstract: The dental identification of edentulous individuals can be challenging based on the lack of antemortem materials and unique features visible on radiographs. The constant resorption of the alveolar ridges further complicates the process. The purpose of this study was to quantify the error rate and reliability of dental identifications based on a comparison of synthesized antemortem and postmortem radiographs of edentulous individuals. Ten observers examined ten cases on two occasions and reported dichotomous and conclusion level decisions. These were analyzed using Kappa and Receiver Operator Characteristics. The mean area under the curve was 0.75 and the mean sensitivity was 0.57 and specificity was 0.83. These results suggest that dental identifications of edentulous individuals using radiographs alone have a high error rate and should be dual reported. These data add further weight to the argument that all dental prostheses should be labeled.     

Identification from Chest X‐Rays: Reliability of Bone Density Patterns of the Humerus*

Abstract: A critical review of Kahana and Hiss’ study on identification from bone trabecular pattern and a test of their method conducted on the humerus are presented. Bone trabecular pattern was studied through the generation of a numerical file representing the gray scale. Using the correlation coefficient, several pairwise comparisons between numerical files were performed. The test gave nearly 30% of incorrect exclusions (the method did not recognize couples of radiographs belonging to the same subject) and 50% of misidentifications (the method recognized couples of radiographs belonging to different subjects, as belonging to the same subject); therefore, this research shows that at the present time, it is not possible to safely quantify identification through bone density patterns, of the proximal humerus taken from thoracic X‐rays. Thus, an “easy”—but dangerous—use of trabecular density patterns on this specific type of radiogram as an identification method should be currently avoided.     

Preliminary Validation of Handheld X‐Ray Fluorescence Spectrometry: Distinguishing Osseous and Dental Tissue from Nonbone Material of Similar Chemical Composition

One of the tasks of a forensic anthropologist is to sort human bone fragments from other materials, which can be difficult when dealing with highly fragmented and taphonomically modified material. The purpose of this research is to develop a method using handheld X‐ray fluorescence (HHXRF) spectrometry to distinguish human and nonhuman bone/teeth from nonbone materials of similar chemical composition using multivariate statistical analyses. The sample materials were derived primarily from previous studies: human bone and teeth, nonhuman bone, nonbiological materials, nonbone biological materials, and taphonomically modified materials. The testing included two phases, testing both the reliability of the instrument and the accuracy of the technique. The results indicate that osseous and dental tissue can be distinguished from nonbone material of similar chemical composition with a high degree of accuracy (94%). While it was not possible to discriminate rock apatite and synthetic hydroxyapatite from bone/teeth, this technique successfully discriminated ivory and octocoral.     

Clinically Detectable Dental Identifiers Observed in Intra‐oral Photographs and Extra‐oral Radiographs,Validated for Human Identification Purposes

Screening the prevalence and pattern of dental identifiers contributes toward the process of human identification. This research investigated the uniqueness of clinical dental identifiers in photographs and radiographs. Panoramic and lateral cephalometric radiographs and five intra‐oral photographs of 1727 subjects were used. In a target set, two observers examined different subjects. In a subset, both observers examined the same subjects (source set). The distance between source and target subjects was quantified for each identifier. The percentage of subjects in the target set being at least as close as the correct subject was assessed. The number of molars (34.6%), missing teeth (42%), and displaced teeth (59.9%) were the most unique identifiers in photographs and panoramic and lateral cephalometric radiographs, respectively. The pattern of rotated teeth (14.9%) was the most unique in photographs, while displaced teeth was in panoramic (37.6%) and lateral cephalometric (54.8%) radiographs. Morphological identifiers were the most unique, highlighting their importance for human identifications.     

Root Morphology and Anatomical Patterns in Forensic Dental Identification: A Comparison of Computer‐Aided Identification with Traditional Forensic Dental Identification*

Abstract: An online forensic dental identification exercise was conducted involving 24 antemortem–postmortem (AM–PM) dental radiograph pairs from actual forensic identification cases. Images had been digitally cropped to remove coronal tooth structure and dental restorations. Volunteer forensic odontologists were passively recruited to compare the AM–PM dental radiographs online and conclude identification status using the guidelines for identification from the American Board of Forensic Odontology. The mean accuracy rate for identification was 86.0% (standard deviation 9.2%). The same radiograph pairs were compared using a digital imaging software algorithm, which generated a normalized coefficient of similarity for each pair. Twenty of the radiograph pairs generated a mean accuracy of 85.0%. Four of the pairs could not be used to generate a coefficient of similarity. Receiver operator curve and area under the curve statistical analysis confirmed good discrimination abilities of both methods (online exercise = 0.978; UT‐ID index = 0.923) and Spearman’s rank correlation coefficient analysis (0.683) indicated good correlation between the results of both methods. Computer‐aided dental identification allows for an objective comparison of AM–PM radiographs and can be a useful tool to support a forensic dental identification conclusion.     

Infrared Spectra of U.S. Automobile Original Finishes (Post – 1989). VIII: In Situ Identification of Bismuth Vanadate Using Extended Range FT‐IR Spectroscopy,Raman Spectroscopy,and X‐Ray Fluorescence Spectrometry

Chrome Yellow (PbCrO₄·xPbSO₄) was a common pigment in U.S. automobile OEM finishes for more than three decades, but in the early 1990s its use was discontinued. One of its main replacements was Bismuth Vanadate (BiVO₄·nBi₂MoO₆, n = 0–2), which was commercially introduced in 1985, as this inorganic pigment also produces a very bright hue and has excellent outdoor durability. This paper describes the in situ identification of Bismuth Vanadate in automotive finishes using FT‐IR and dispersive Raman spectroscopy and XRF spectrometry. Some differentiation of commercial formulations of this pigment is possible based on far‐infrared absorptions, Raman data, and elemental analysis. The spectral differences arise from the presence or absence of molybdenum, the use of two crystal polymorphs of BiVO₄, and differences in pigment stabilizers. Bismuth Vanadate is usually not used alone, and it is typically found with Isoindoline Yellow, hydrous ferric oxide, rutile, Isoindolinone Yellow 3R, or various combinations of these.     

Human Identification Using Automatic and Semi‐Automatically Detected Facial Marks

Continuing advancements in the field of digital cameras and surveillance imaging devices have led law enforcement and intelligence agencies to use analysis of images and videos for the investigation and prosecution of crime. When determining identity from photographic evidence, forensic analysts perform comparison of visible facial features manually, which is inefficient. In this study, we will address research efforts to use facial marks as biometric signatures to distinguish between individuals. We propose two systems to assist forensic analysts during photographic comparison: an improved multiscale facial mark system in which facial marks are detected automatically, and a semi‐automatic facial mark system that integrates human knowledge within the improved multiscale facial mark system. Experiment results employ a high‐resolution time‐elapsed dataset acquired at the University of Notre Dame between 2009 and 2011. The results indicate that the geometric distributions of facial mark patterns can be used to distinguish between individuals.     

The Use of Near‐Infrared Photography to Image Fired Bullets and Cartridge Cases

An imaging technique that is capable of reducing glare, reflection, and shadows can greatly assist the process of toolmarks comparison. In this work, a camera with near‐infrared (near‐IR) photographic capabilities was fitted with an IR filter, mounted to a stereomicroscope, and used to capture images of toolmarks on fired bullets and cartridge cases. Fluorescent, white light‐emitting diode (LED), and halogen light sources were compared for use with the camera. Test‐fired bullets and cartridge cases from different makes and models of firearms were photographed under either near‐IR or visible light. With visual comparisons, near‐IR images and visible light images were comparable. The use of near‐IR photography did not reveal more details and could not effectively eliminate reflections and glare associated with visible light photography. Near‐IR photography showed little advantages in manual examination of fired evidence when it was compared with visible light (regular) photography.     

Scanning Electron Microscopy–Energy‐Dispersive X‐Ray (SEM/EDX): A Rapid Diagnostic Tool to Aid the Identification of Burnt Bone and Contested Cremains

This study investigates the use of Scanning electron microscopy–energy‐dispersive X‐ray (SEM‐EDX) as a diagnostic tool for the determination of the osseous origin of samples subjected to different temperatures. Sheep (Ovis aries) ribs of two experimental groups (fleshed and defleshed) were burned at temperatures of between 100°C and 1100°C in 100°C increments and subsequently analyzed with the SEM‐EDX to determine the atomic percentage of present elements. Three‐factor ANOVA analysis showed that neither the exposure temperature, nor whether the burning occurred with or without soft tissue present had any significant influence on the bone's overall elemental makeup (p > 0.05). The Ca/P ratio remained in the osseous typical range of between 1.6 and 2.58 in all analyzed samples. This demonstrates that even faced with high temperatures, the overall gross elemental content and atomic percentage of elements in bone remain stable, creating a unique “fingerprint” for osseous material, even after exposure to extreme conditions.     

A Novel Approach to the Examination of Soil Evidence: Mineral Identification Using Infrared Microprobe Analysis*

Abstract: Identification of minerals using the infrared microprobe with a diamond internal reflection objective is a rapid and reliable method for forensic soil examinations. Ninety‐six mineral varieties were analyzed, and 77 were differentiated by their attenuated total reflection (ATR) spectra. Mineral grains may be mounted in oil for conventional polarized light microscope characterization and their ATR spectrum obtained with little or no interference by the liquid. This infrared microprobe method can be used to identify silicates, phosphates, nitrates, carbonates, and other covalent minerals; however, ionic minerals, metal oxide and sulfide minerals, and minerals with refractive indexes greater than diamond do not produce identifiable spectra, but the lack of a spectrum or one with high absorbance values does provide useful information. This research demonstrates the overall utility that infrared microprobe analysis brings mineral identification in soil evidence.     

Development of a Protein‐based Human Identification Capability from a Single Hair

Shed human hair (lacking root nuclear DNA) frequently contributes important information to forensic investigations involving human identification. Detection of genetic variation observed in amino acid sequences of hair proteins provides a new suite of identity markers that augment microscopic hair analysis and mitochondrial DNA sequencing. In this study, a new method that completely dissolves single hairs using a combination of heat, ultrasonication, and surfactants was developed. Dissolved proteins were digested and genetically variant peptide (GVP) profiles were obtained for single hairs (25 mm) via high‐resolution nanoflow liquid chromatography‐based mass spectrometry and a novel exome‐driven bioinformatic approach. Overall, 6519 unique peptides were identified and a total of 57 GVPs were confirmed. Random match probabilities ranged between 2.6 × 10^?2 and 6.0 × 10^?9. The new bioinformatic strategy and ability to analyze GVPs in forensically relevant samples sizes demonstrate applicability of this approach to distinguish individuals in forensic contexts.     

The Potential of X‐Ray Diffraction in the Analysis of Burned Remains from Forensic Contexts*

Abstract: In view of the difficulties in extracting quantitative information from burned bone, we suggest a new and accurate method of determining the temperature and duration of burning of human remains in forensic contexts. Application of the powder X‐ray diffraction approach to a sample of human bone and teeth allowed their microstructural behavior, as a function of temperature (200–1000°C) and duration of burning (0, 18, 36, and 60 min), to be predicted. The experimental results from the 57 human bone sections and 12 molar teeth determined that the growth of hydroxylapatite crystallites is a direct and predictable function of the applied temperature, which follows a nonlinear logistic relationship. This will allow the forensic investigator to acquire useful information about the equilibrium temperature brought about by the burning process and to suggest a reasonable duration of fire exposure.     

Proteolytic‐Based Method for the Identification of Human Growth Hormone*

Abstract: Human growth hormone (HGH) is a relatively small protein consisting of 191 amino acids and has an average mass of 22,125 amu. The forensic analysis of proteins such as HGH must meet the analytical sufficiency requirements for the laboratory and consists of a binary approach. A suspected sample is analyzed as the whole protein for retention time and mass determination using high performance liquid chromatography equipped with a photodiode array and liquid chromatography mass spectrometry. Further fragmentation of the protein using a proteolytic enzyme adds another dimension to the specificity of the analysis. Porcine trypsin digests proteins in a very predictable manner and yields peptide fragments of the original protein that can be used as a means for fingerprinting the larger biomolecule. In silico, or theoretical, digestion of HGH by trypsin yields 21 peptides ranging in size from 1 to 23 amino acids in length. The larger fragments containing higher numbers of amino acids give more specificity to identifying a protein based on a fragment produced by the digestion of trypsin. Herein, the analysis of HGH using a proteolytic approach is presented that meets the Scientific Working Group for the Analysis of Seized Drugs (SWGDRUG) recommendations for the identification of unknown substances.     

Identification of Different Forms of Cocaine and Substances Used in Adulteration Using Near‐infrared Raman Spectroscopy and Infrared Absorption Spectroscopy

Identification of cocaine and subsequent quantification immediately after seizure are problems for the police in developing countries such as Brazil. This work proposes a comparison between the Raman and FT‐IR techniques as methods to identify cocaine, the adulterants used to increase volume, and possible degradation products in samples seized by the police. Near‐infrared Raman spectra (785 nm excitation, 10 sec exposure time) and FT‐IR‐ATR spectra were obtained from different samples of street cocaine and some substances commonly used as adulterants. Freebase powder, hydrochloride powder, and crack rock can be distinguished by both Raman and FT‐IR spectroscopies, revealing differences in their chemical structure. Most of the samples showed characteristic peaks of degradation products such as benzoylecgonine and benzoic acid, and some presented evidence of adulteration with aluminum sulfate and sodium carbonate. Raman spectroscopy is better than FT‐IR for identifying benzoic acid and inorganic adulterants in cocaine.     

Identification of Human Remains by DNA Analysis of the Gastrointestinal Contents of Fly Larvae

Dipterous fly larvae (maggots) are frequently collected from a corpse during a criminal investigation. Previous studies showed that DNA analysis of the gastrointestinal contents of maggots might be used to reveal the identity of a victim. However, this approach has not been used to date in legal investigations, and thus its practical usefulness is unknown. A badly burned body was discovered with its face and neck colonized by fly larvae. Given the condition of the body, identification was not possible. Short tandem repeat (STR) typing was performed using the gastrointestinal contents of maggots collected from the victim and was compared to STR profiles obtained from the alleged father. The probability of paternity was 99.685%. Thus, this comparative DNA test enabled the conclusive identification of the remains. This is the first reported case of analysis of human DNA isolated from the gastrointestinal tract of maggots used to identify a victim in a criminal case.     

A Review of Bomb Pulse Dating and its Use in the Investigation of Unidentified Human Remains

In cases where there is limited antemortem information, the examination of unidentified human remains as part of the investigation of long-term missing person's cases is a complex endeavor and consequently requires a multidisciplinary approach. Bomb pulse dating, which involves the analysis and interpretation of 14C concentration, is one technique that may assist in these investigations by providing an estimate of year of birth and year of death. This review examines the technique of bomb pulse dating and its use in the identification of differentially preserved unknown human remains. Research and case studies implementing bomb pulse dating have predominantly been undertaken in the Northern Hemisphere and have demonstrated reliable and accurate results. Limitations were, however, identified throughout the literature. These included the small sample sizes used in previous research/case studies which impacted on the statistical significance of the findings, as well as technique-specific issues. Such limitations highlight the need for future research.     

Forensic Comparison of Automotive Aluminum Wheel Fragments Using Synchrotron Radiation X‐ray Fluorescence with 18‐ and 116‐keV Excitation X‐rays

The performance of synchrotron radiation X‐ray fluorescence (SR‐XRF) spectrometry for nondestructive discrimination of small fragments of automotive aluminum wheels was studied. Fragments (< 500 × 500 μm²) of 45 kinds of wheels were first analyzed by scanning electron microscopy–energy dispersive spectroscopy (SEM‐EDS) and then by SR‐XRF. Despite the Mg/Al intensity ratio being a useful identification index, SEM‐EDS was not efficient enough because of the absence of other meaningful indicators of comparison. Conversely, pairwise comparison was conducted and a 92.9% identification was achieved via SR‐XRF using 18‐keV X‐rays. Trace heavy elements in the high‐energy region were detected by SR‐XRF using 116‐keV X‐rays, and an 82.9% identification was obtained. Combined use of 18‐ and 116‐keV X‐rays allowed 98.2% identification. Hence, SR‐XRF is a powerful tool for nondestructive discrimination of automotive aluminum wheels with high precision using trace elements in a wide energy region.     

Development of a Computer‐Assisted Forensic Radiographic Identification Method Using the Lateral Cervical and Lumbar Spine

Medical examiners and coroners (ME/C) in the United States hold statutory responsibility to identify deceased individuals who fall under their jurisdiction. The computer‐assisted decedent identification (CADI) project was designed to modify software used in diagnosis and treatment of spinal injuries into a mathematically validated tool for ME/C identification of fleshed decedents. CADI software analyzes the shapes of targeted vertebral bodies imaged in an array of standard radiographs and quantifies the likelihood that any two of the radiographs contain matching vertebral bodies. Six validation tests measured the repeatability, reliability, and sensitivity of the method, and the effects of age, sex, and number of radiographs in array composition. CADI returned a 92–100% success rate in identifying the true matching pair of vertebrae within arrays of five to 30 radiographs. Further development of CADI is expected to produce a novel identification method for use in ME/C offices that is reliable, timely, and cost‐effective.