Scanning electron microscopy analysis of experimental bone hacking trauma

The authors report on their macro- and microscopy study of bone lesions made by a sharp force instrument (a single blade knife), and a sharp-blunt instrument classified as a chopping weapon (a hatchet). The aim of this work was to attempt to identify the instrument by analyzing the general class characteristics of the cuts. Each weapon was used on human bones. The results indicate that macroscopic analysis is more problematic. The microscopic analysis assessed that characteristics examined were effective in distinguishing sharp from sharp-blunt injury to the bone. The microscope facilitates analysis unachievable with macroscopic methods, some three-dimensional characteristics not visible to the naked eye being clearly defined with its use. Emphasis has been placed on the value of SEM as an anthropologist's tool in bone lesion injuries.     

Macroscopic characteristics of hacking trauma

Hacking trauma is often encountered in forensic cases, but little experimental research has been conducted that would allow for the recognition of wounds caused by specific weapon types. In this paper, we report the results of a hacking trauma caused by machete, cleaver, and axe weapons and the characteristics of each weapon type on bone. Each weapon type was employed in multiple trials on pig (Sus scrofa) bones and then the wounds were examined macroscopically for several characteristics that serve to differentiate the weapons.     

Scanning electron microscopy of incinerated teeth

The aim of the present scanning electron microscopy study was to document the nature of morphologic changes occurring in human enamel and dentin subjected in vitro to temperatures in the range of 200-1,000 degrees C for variable times. The results of the investigation confirm that human enamel and dentin remain microscopically identifiable after incineration at 1000 degrees C. Furthermore, these tissues remain identifiable after incineration at 1,000 degrees C for periods greater than 3 h. No consistent or reliable differences in morphology could be detected in enamel or mineralized dentin incinerated in the temperature range 200-600 degrees C. Temperature-dependent changes involving the predentin zone were observed. Following incineration at 800 degrees C for over 3 h and at 1,000 degrees C for 3 h, a metamorphosis of enamel and dentin into a globular form was observed.     

Microscopic characteristics of hacking trauma

The purpose of this study was to determine if it is possible to associate machetes, axes, and cleavers with the microscopic parallel striations they leave on the cut surfaces of the bone. Hacking trauma was experimentally inflicted on pig bones using machetes, axes, and cleavers. Negative impressions of both the cut surfaces of the bone and the weapon blades were analyzed using scanning electron microscopy. The results of this investigation indicate that it is possible to correlate a class of hacking weapons to trauma inflicted on bone by these weapons.     

Scanning electron microscopic analysis of skin resolution as an aid in identifying trauma in forensic investigations

The forensic investigator is frequently confronted with cases that present with wounds and blunt force trauma. Presently, the forensic investigator depends upon previous experience and further investigative deduction of the crime scene to analyze these injuries. Although not readily apparent to the naked eye, many skin tissue injuries can be visualized with scanning electron microscopy (SEM). This study was designed to establish skin trauma resolution using SEM in various skin preparations. Tissue trauma was induced on leather, preserved skin, fresh skin, and living skin using dies of varying thread size. Calibrated pressure forces in pounds per square inch (psi) were applied and impressions made using vinyl polysiloxane. Positive replicas of the tissues were prepared for SEM using isocyanate resin. After sputter coating the cast with 35 nm of gold-palladium, electron micrographs were generated using a Jeol JSM-5310LV scanning electron microscope. To establish resolution, thread widths of 52, 104, and 208 threads per inch (tpi) and trauma forces of 150, 200, and 250 psi were used to produce the impressions. Microgrooves that were identified on the die threads were analyzed. The optimum pressure for resolution studies was 150 psi using the 52 tpi die on the leather sample (4.67 +/- 0.88 microm, p = 0.046 and 0.025, respectively, by ANOVA). The resolution was compared to that of leather using preserved, fresh, and living skin. The resolution in preserved and fresh skin was less than for leather (9.00 +/- 1.73 and 10.5 +/- 4.5 versus 4.67 +/- 0.88 microm, p = 0.09 and p = 0.20, respectively). Living skin resolution was 3 microm at 52 tpi and 100 psi. Various implements of blunt force trauma were also examined using the leather sample. Time after trauma resolution was examined at 0 (3 microm), 5 (6 microm), 10 (8 microm), and 20 (9 microm) min in living tissue. A comparison between the microgrooves on the die replicas and the tissue trauma impressions revealed striking agreement for both linearity and resolution. Analysis of the microgrooves suggests that discrete morphological characteristics are seen in skin tissue traumas. This method could expand the tools available for the forensic investigation of blunt force trauma.     

Scanning electron microscopy determination of string mozzarella cheese in gastric contents

As part of a suspected homicide investigation, a sampling of the gastric contents from the victim was forwarded to the U.S. Food and Drug Administration's Forensic Chemistry Center (FCC) for analysis of specific, selected components. The victim was known to have consumed string mozzarella cheese, as a snack, less than 24 h before his disappearance and the subsequent discovery of the body. The investigation sought to confirm or dismiss speculation the victim may have been fed a meal or eaten additional food prior to his death. Analysis of the stomach contents involved examination by stereoscopic light microscopy (SLM) and isolation, processing, and analysis of suspect materials by scanning electron microscopy (SEM). Several wax-like, off-white to cream-colored objects were noted by SLM examination and removed from the gastric contents. Through a series of fixation, sectioning, drying, and coating steps, these objects were prepared for analysis by SEM. Comparison of the suspect material with laboratory control string mozzarella cheese showed excellent correlation between the analyzed samples, confirming the suspect material from the stomach contents as string mozzarella cheese.     

Scanning electron microscopy as an auxiliary method in the study of exhumed bones

Scanning electron microscopy (SEM) has been used in forensic science in many ways. The reports of cases in which SEM has been used as an auxiliary method in the investigation of exhumed bones are rare. In this article, we report an exhumation that was made to determine if a seized weapon could have been used in a homicide. We used SEM to analyze a fracture in the interior of the skull of the victim. The findings described in this article showed us that it is possible to develop new researches in this field.     

Quantitative analysis of sharp-force trauma: an application of scanning electron microscopy in forensic anthropology.

Scanning electron microscopy (SEM) has occasionally been used by anthropologists and forensic scientists to look at morphological characteristics that certain implements leave on bone. However, few studies have addressed techniques or protocols for assessing quantitative differences between tool marks on bone made by different bladed implements. In this study, the statistical variation in cut mark width was examined between control and test samples on bone using a scalpel blade, paring knife, and kitchen utility knife. Statistically significant differences (p < .0005) were found between cut marks made by the same knife under control and test conditions for all three knife types used in the study. When the control sample and test samples were examined individually for differences in mean variation between knife types, significant differences were also found (p < .0005). While significant differences in cut mark width were found, caution should be used in trying to classify individual cut marks as being inflicted by a particular implement, due to the overlap in cut mark width that exists between different knife types. When combined, both quantitative and qualitative analyses of cut marks should prove to be more useful in trying to identify a suspect weapon. Furthermore, the application of SEM can be particularly useful for assessing many of these features.     

Scanning electron microscopy: application in the identification of diatoms in cases of drowning.

A simple and rapid method is described for processing organ and water samples for the identification of diatoms so that they can be studied and recorded for taxonomic and diagnostic purposes. Samples are treated with concentrated nitric acid; the fluid obtained is centrifuged, and its sediment is dried, coated, and examined under the scanning electron microscope. The method does not alter the morphology of diatoms and allows the study of freshwater and seawater species present in the organs of bodies found in the water, making possible the diagnosis of drowning under specific conditions.     

Forensic analysis of soil and sediment traces by scanning electron microscopy and energy-dispersive X-ray analysis: an experimental investigation

This paper reports the results of a series of experiments carried out to determine the precision of soil trace comparisons based on elemental peak height ratios determined by energy-dispersive X-ray analysis (EDXRA) in a variable pressure scanning electron microscope (VP-SEM). Experiments were conducted on 'bulk' soil aggregates, ground powders prepared from the <150 microm soil fractions and on smears of both the bulk soil and <150 microm material placed on cotton cloth. X-ray count data were obtained using area scans and spot analyses at different magnifications. The effects on elemental peak height ratios of varying the SEM chamber pressure, beam spot size, emission current and accelerating voltage were also examined. The peak height ratios for oxygen, silicon, aluminium, potassium, calcium and iron were found to show little variation as a function of chamber pressure, spot size and emission current over the ranges examined, but a strong dependency on accelerating voltage was observed. Within-sample variation in results, expressed by the percentage coefficient of variation, was found to be lowest for area scan analyses of the ground <150 microm fractions and greatest for the spot analyses of the bulk soil aggregates and the <150 microm fractions. We conclude that comparison of elemental peak height ratios determined by EDXRA can be a useful tool for rapid screening of soil samples, especially when combined with investigation of other attributes of the soil traces such as colour, fabric and the composition, shapes and surface textures of individual particles or aggregates within the soil traces. If sufficient material is available and can be readily separated without contamination or loss, higher resolution and more precise elemental data should be obtained by methods such as inductively coupled plasma atomic-emission spectrometry (ICP-AES) or mass-spectrometry (ICP-MS).     

Incised wound margins caused by steel blades. Scanning electron microscopy to determine wound direction

The aim of this paper has been to check the diagnostic ability of Scanning Electron Microscopy (S.E.M.) to determine wound direction. We have studied incised wound margins caused by steel blades in human skin samples from cadavers by S.E.M. It was found that a lateral accessory tail existed in 65% of the entrance edges. This finding was recorded in all the cases when wound direction was perpendicular to Lancer's lines of the skin area in which the wound was located. No similar finding in the exit tails was noticed. The depth and length of the wound margins was less accurate an index of the wound direction than a lateral accessory tail. We think that this finding could be useful in determining the entrance margin in incised wounds.     

Scanning electron microscope study of the lung in drowning

The authors describe the results obtained using the scanning electron microscope (SEM) to study drowned lungs and control lungs. Results are compared with a previous study carried out using laboratory animals. The results from both studies are analogous, in the absence of chronic pulmonary emphysema. The diagnosis of chronic pulmonary emphysema is simple with the SEM even when dealing with autoptic material obtained 24 to 48 h after death, but the diagnosis of drowning becomes difficult in lungs with chronic emphysema.     

Discriminant analysis of the mandible for sex determination

Seventeen measuring parameters were used to characterize 197 mandibles (109 males, 88 females) taken from the corpses of people 20-80 years of age from the Rhine-Main-Neckar area. The representative measuring parameters and discriminating functions of intact lower jaws and lower jaw fragments of most frequent fracture types were determined in this practice group by means of discriminating analysis. In the present research material it was possible to determine sex accurately from an intact lower jaw bone in 82.6% (m) and 79.5% (f) of the cases. Furthermore, our results show clearly that sex may even be determined from lower jaw fragments. The classification was correct, depending on the type of fragment, in 72.5%-81.7% (m) and 71.6%-79.5% (f) of the cases.     

Adjunctive use of scanning electron microscopy in bite mark analysis: a three-dimensional study

An examination of a case in which adjunctive use of scanning electron microscopy (SEM) demonstrated the presence of unusual three-dimensional characteristics in a bite mark. Despite the fact that many bite marks do not show "depth," demonstration of the presence of this third dimension can produce significant data for evidentiary purposes. In some instances, these data may transform what seem to be class characteristics of a bite mark into individual characteristics and thus impart much more uniqueness to the evidence. Because of the high level of resolution and magnification of SEM, some three-dimensional characteristics not visible to the eye can be shown quite clearly by its use. Emphasis will be given to the value of SEM as a tool of the forensic odontologist in bite mark analysis.     

The ultrastructure of the electric burn in man: a transmission electron microscopy-scanning electron microscopy study

The authors describe the ultrastructural aspects of the electric burn observed with the transmission electron microscope (TEM) and the scanning electron microscope (SEM). With the TEM the most apparent feature is the homogenization and arrangement of the filamentous cytoplasmic material in elongated and parallel bundles together with elongation of the nuclei and junctional structures. SEM studies present a three-dimensional vision of the filamentous material that confirms and clarifies the cellular deformation as a whole. An hypothesis is advanced suggesting that the phenomenon is due to the effects produced by the extremely rapid dehydration of the tissue or that the electric current and Joule effect can act upon the filaments and contractile proteins, causing the cell deformation.     

Chemical trace XRF analysis to detect sharp force trauma in fresh and burned bone

Forensic anthropologists may not always be able to differentiate heat-induced fractures from fractures with other aetiologies, namely sharp force traumas, with clear nefarious impact on medico-legal conclusions. The objective of this study was to experimentally investigate if blade chemical traces are transferred to defleshed bone tissue and if they remain there after a burning event. This was accomplished by prompting sharp force traumas in 20 macerated fresh pig ribs with five different instruments, namely a stainless steel knife, an artisanal knife and a ceramic kitchen knife, a small axe and a large axe. Another pig rib was used as control, not being subjected to any trauma. All instruments were probed by X-ray fluorescence (XRF) to establish the composition of each blade. Bone samples, both pre-burned and post-burned (at 500 °C, 700 °C, 900 °C and 1100 °C), were then probed by XRF. All sharp force instruments left detectable chemical traces on pre-burned bone, although not in all samples. Furthermore, traces were still detected after experimental burning in most cases. Potentially, XRF can provide relevant information about the aetiology of fractures in burned and unburned bones, although the effect of soft tissues and diagenesis must still be investigated.