Evaluation of the Forensic Utility of Scanning Electron Microscopy‐Energy Dispersive Spectroscopy and Laser Ablation‐Inductively Coupled Plasma‐Mass Spectrometry for Printing Ink Examinations

Improvements in printing technology have exacerbated the problem of document counterfeiting, prompting the need for analytical techniques that better characterize inks for forensic analysis and comparisons. In this study, 319 printing inks (toner, inkjet, offset, and Intaglio) were analyzed directly on the paper substrate using scanning electron microscopy‐energy dispersive spectroscopy (SEM‐EDS) and Laser Ablation‐Inductively Coupled Plasma‐Mass Spectrometry (LA‐ICP‐MS). As anticipated, the high sensitivity of LA‐ICP‐MS pairwise comparisons resulted in excellent discrimination (average of ~ 99.6%) between different ink samples from each of the four ink types and almost 100% correct associations between ink samples known to originate from the same source. SEM‐EDS analysis also resulted in very good discrimination for different toner and intaglio inks (>97%) and 100% correct association for samples from the same source. SEM‐EDS provided complementary information to LA‐ICP‐MS for certain ink types but showed limited utility for the discrimination of inkjet and offset inks.     

The Effects of Environmental Exposure on the Optical,Physical, and Chemical Properties of Manufactured Fibers of Natural Origin

Manufactured fibers derived from natural origins include viscose rayon, azlon, and polylactic acid (PLA). A 2‐year study was conducted to document any changes these fibers undergo as a result of exposure to various environmental conditions. Fabric swatches representing each fiber type were exposed to freshwater, saltwater, heat, cold, ultraviolet light, or composter conditions. Fibers from the swatches were periodically analyzed using polarized light microscopy and Fourier transform infrared microspectroscopy. Fiber solubility and melting‐point behavior were measured every 6 months. Except for the complete degradation of viscose rayon in the composter, saltwater, and freshwater environs, no changes in the optical properties, infrared spectra, solubility, or melting points of the remaining fibers in any of the environments were observed. However, microscopic morphological changes were observed in fibers from two azlon swatches submerged in freshwater and saltwater, two PLA swatches exposed to ultraviolet light, and two viscose rayon swatches exposed to ultraviolet light.     

The Effect of Skin Debris on Gunshot Residue Sampling and Detection

These experiments were designed to determine whether skin debris (desquamated epithelial cells and apparent skin oils) affects gunshot residue (GSR) particle detection on the sticky tape lift samples prepared for scanning electron microscopy (SEM). A dabbing experiment showed that GSR particles accumulate not only on the adhesive surface of the sampler, but also on the epithelial cell surfaces. Samplers were loaded with target GSR followed by dabbing 30 times on the back of a hand. Backscatter electron images were taken at 20 kV and for some at 30 kV of the same areas. The samplers were then treated with a sodium/calcium hypochlorite solution (bleach) to remove skin debris and again imaged in the SEM. Comparison of these images shows more GSR particles will likely be revealed at 30 kV than 20 kV and more particles revealed by the bleach treatment in an automated SEM system.     

Atomic force microscopy as a biophysical tool for nanoscale forensic investigations

The atomic force microscope (AFM) has found its way to the arsenal of tools available to the forensic practitioner for the analysis of samples at the nano and microscales. As a non-destructive probing tool that requires minimal sample preparation, the AFM is very attractive, particularly in the case of minimal or precious sample. To date, the use of the AFM has primarily been in the arena of imaging where it has been complementary to other microscopic examination tools. Forensic applications in the visual examination of evidence such as blood stains, questioned documents, and hair samples have been reported. While a number of reviews have focused on the use of AFM as an imaging tool for forensic analyses, here we not only discuss these works, but also point to a versatile enhancement in the capabilities of this nanoscale tool – namely its use for force spectroscopy. In this mode, the AFM can determine elastic moduli, adhesion forces, energy dissipation, and the interaction forces between cognate ligands, that can be spatially mapped to provide a unique spatial visualization of properties. Our goals in this review are to provide a context for this capability of the AFM, explain its workings, cover some exemplary works pertaining to forensic sciences, and present a critical analysis on the advantages and disadvantages of this modality. Equipped with this high-resolution tool, imaging and biophysical analysis by the AFM can provide a unique complement to other tools available to the researcher for the analysis and characterization of forensic evidence.     

Persistence of cellular material after exposure to water

Disposing of items of forensic relevance in bodies of water is one countermeasure offenders can use to avoid detection. The impact of immersion in water has been explored for blood, saliva, and semen; however, few studies have assessed touch DNA. Here we report on the effect of exposure to water on the persistence of touch DNA over prolonged periods of time. To evaluate the persistence of cells from touch DNA, after water exposure, three substrates and two water types were tested: plastic, metal, and ceramic, submerged into seawater or tap water. Diamond™ Nucleic Acid Dye was used to stain cells deposited by touch. Cell counts before and after water exposure were compared to investigate cell loss over time, ranging from 6 hours to 5 days. A logarithmic increase in the percent of cells lost was observed over time when the data for substrate and water type conditions were combined. Substrate type influenced the persistence of cells, with the metal substrate retaining cells longer than plastic or ceramic. The influence of water type appeared dependent on the substrate, with varied cell persistence on metal whereas plastic and ceramic recorded similar cell loss over time between water types. The ability to visualize cells after exposure to water could assist in triaging evidence within operational forensic laboratories and allow for targeted sampling. This proof-of-concept study demonstrated that greater than 50% of cells can persist on various items submerged in aqueous environments for at least 5 days, highlighting the possibility for downstream DNA testing.     

UBM、SL-OCT及房角镜检查前房角结构的比较

目的 探讨超声生物显微镜（ultrasound biomicroscope,UBM）、裂隙灯前节光学相干断层成像（slit lamp optical coherence tomography,SL-OCT）与房角镜检查房角后退和房角关闭的一致性. 方法 在同一暗室中对被检查眼上方、下方、颞侧、鼻侧4个方位进行UBM、SL-OCT及房角镜检查,记录前房角情况,应用x2检验和Kappa检验对3种检查方法就房角关闭、房角后退的检查结果一致性进行比较. 结果 应用UBM、SL-OCT及房角镜检查判断房角关闭和房角后退,3种检查方法差异无统计学意义（P＞0.05）,UBM与房角镜检查房角后退结果一致性更好（Kappa值0.882）,而SL-OCT与房角镜检查房角后退结果一致性一般（Kappa值0.624）. 结论 以房角镜检查作为标准,对房角后退的判断UBM优于SL-OCT;对房角关闭的判断,UBM、SL-OCT及房角镜检查一致性均良好.     

Using Raman Spectroscopy to solve crime: inks, questioned documents and fraud

Raman microscopy is becoming a tool of major importance in forensic analysis, particularly of drugs and explosives. It is a non-invasive, non-destructive chemical probe allowing samples to be examined in their entirety without any preparation. This paper demonstrates the use of the technique as a general tool for inks analysis. Furthermore, it addresses two important issues that historically have been extremely difficult for the professional document examiner, namely, comparison of black ballpoint inks and the chronological sequencing of crossed ink lines. We show that Raman can successfully distinguish between a representative sample of commercially available black ballpoint inks. This data has been converted into a database for future reference. A method for chronological sequencing of crossed ink lines has been developed using confocal Raman microscopy. Case study work has shown the feasibility of this approach.     

Standard 28-cycle amplification of X/Y-FISH labelled epithelial cells for DNA profiling

The examination of sexual assault evidence frequently involves the analysis of samples that comprise mixtures of male and female cells. Separating male and female cells benefits analysis as the results are more likely to be simplified into profiles from single contributors. Some separation methods have focussed on separation of sperm from epithelial cells, but samples without sperm also require separation (vasectomised males, licked skin, etc.). X/Y chromosome FISH labelling when combined with laser micro-dissection (LMD) is a reliable method to separate male and female epithelial cells, but has mostly been combined with increased cycle PCR to create DNA profiles, limiting its use in many forensic laboratories. This study aimed to determine the limits of cell numbers collected by LMD for standard 28-cycle DNA profiling, and to test the effects, if any, on stochastic variation normally caused by sampling effects. Male and female epithelial cells were stained using the Vysis CEP X/Y DNA Probe kits, and collected using a Leica LMD6000. DNA was extracted and amplified by the ESR in-house one-tube method, using standard 28-cycle PCR with the AmpFISTR Identifiler™ (Applied Biosystems) multiplex kit. Full IdentifilerTM DNA profiles were produced using standard 28-cycle PCR, and partial profiles suitable for submission were produced from even relatively low numbers of cells collected. Profiling results were compared with low-copy number PCR on low numbers of cells stained and collected in the same manner, and the observed effects on heterozygote balance are discussed.     

SEM and Stereomicroscopic Analysis of Cut Marks in Fresh and Burned Bone

This study documents the prevalence of cut mark characteristics in fresh and burned domestic pig ribs. Stab wounds from single edge serrated and smooth‐edged knives were inflicted in the vertebral and sternal regions of each fresh rib. Each rack of ribs was then divided into vertebral and sternal units. Vertebral units were defleshed and their associated cut marks were examined using a stereomicroscope. Sternal units were burned in an outdoor fire pit and examined with the addition of a scanning electron microscope (SEM). Linear cuts, V‐shaped cross‐sections, mounding, hinge fractures, and wastage were all observed on burned ribs. There was an overall decrease in the prevalence of all features (up to a 40% decrease), regardless of knife type, in burned ribs. Striations within cut marks were not observed in either fresh or burned ribs. Oblique faulting and bone lifts could only be observed using the SEM. Mounding and wastage were obliterated during the burning process. Therefore, cut marks in burned bone should ideally be examined for their characteristics utilizing an SEM.     

Lead Theft—A Study of the “Uniqueness” of Lead from Church Roofs

In the United Kingdom, theft of lead is common, particularly from churches and other public buildings with lead roofs. To assess the potential to distinguish lead from different sources, 41 samples of lead from 24 church roofs in Northamptonshire, U.K, have been analyzed for relative abundance of trace elements and isotopes of lead using X‐ray fluorescence (XRF) and inductively coupled plasma mass spectrometry, respectively. XRF revealed the overall presence of 12 trace elements with the four most abundant, calcium, phosphorus, silicon, and sulfur, showing a large weight percentage standard error of the mean of all samples suggesting variation in the weight percentage of these elements between different church roofs. Multiple samples from the same roofs, but different lead sheets, showed much lower weight percentage standard errors of the mean suggesting similar trace element concentrations. Lead isotope ratios were similar for all samples. Factors likely to affect the occurrence of these trace elements are discussed.