Application of Volume‐of‐Fluid Method to Analyze the Viscosity Effect on the Spine Formation of Bloodstains

In bloodstain pattern analysis, the blood droplet volume and surface impact velocity play an important role, and many related experimental studies have been carried out. If an appropriate computational fluid dynamics (CFD) model that could solve bloodstain patterns, especially spine formation bloodstain patterns, can be obtained, the blood droplet volume and impact speeds at various crime scenes can be predicted more accurately. For this purpose, Flow‐3D software using the volume‐of‐fluid method was applied to analyze the behavior of human blood droplets during an impact event, especially focusing on the viscous effect on splashing, which forms the spine which can be used to predict the impact velocity. To obtain a non‐Newtonian viscosity model of blood for a computational fluid dynamic analysis, the venous blood samples of 163 people were tested using a hemorheology instrument. Among the venous blood samples of 163 people, 37 samples for which all blood test results were in a normal range were selected for the non‐Newtonian viscosity modeling. From the CFD analysis, it could be concluded that a non‐Newtonian viscosity model is more appropriate than a constant viscosity model for predicting splashing that forms the spine. The gradient of the non‐Newtonian model at a high shear rate has more of an effect on spine formation than that at a low shear rate. The lowest viscosity with a high velocity at the outer front of the radiating flow plays an important role in forming the splashing pattern.     

The Reliability of Pattern Classification in Bloodstain Pattern Analysis—PART 2: Bloodstain Patterns on Fabric Surfaces,

This study was designed to produce the first baseline measure of the reliability of bloodstain pattern classifications on fabric surfaces. Experienced bloodstain pattern analysts classified bloodstain patterns on pairs of trousers that represented three fabric substrates. Patterns also varied in type (impact, cast‐off, expiration, satellite stains from dripped blood, and transfer) and extent. In addition, case summaries that accompanied each pattern contained contextual cues that either supported the correct answer (i.e., positive bias), were misleading toward an incorrect answer (i.e., negative bias), or contained no directional information (i.e., neutral). Overall, 23% percent of the resulting classifications were erroneous. The majority (51%) of errors resulted from analysts misclassifying satellite stains from dripped blood. Relative to the neutral information, the positive‐bias information increased correct classifications and decreased erroneous classifications, and the negative‐bias information decreased correct classifications and increased erroneous classifications. The implications of these findings for BPA are discussed.     

Quantitative Differentiation of Bloodstain Patterns Resulting from Gunshot and Blunt Force Impacts

Bloodstain pattern analysis (BPA) provides significant evidentiary value in crime scene interpretation and reconstruction. In this work, we develop a quantitative methodology using digital image analysis techniques to differentiate impact bloodstain patterns. The bloodstain patterns were digitally imaged and analyzed using image analysis algorithms. Our analysis of 72 unique bloodstain patterns, comprising more than 490,000 individual droplet stains, indicates that the mean drop size in a gunshot spatter pattern is at most 30% smaller than the mean drop stain size in blunt instrument patterns. In contrast, we demonstrate that the spatial distribution of the droplet stains—their density as a function of position in the pattern—significantly differs between gunshot and blunt instrument patterns, with densities as much as 400% larger for gunshot impacts. Thus, quantitative metrics involving the spatial distribution of droplet stains within a bloodstain pattern can be useful for objective differentiation between blunt instrument and gunshot bloodstain patterns.     

Alteration of expirated bloodstain patterns by Calliphora vicina and Lucilia sericata (Diptera: Calliphoridae) through ingestion and deposition of artifacts

Abstract: Bloodstain pattern analysis can provide insight into a sequence of events associated with a violent crime. However, bloodstain pattern analysis can be confounded by the feeding activity of blow flies. We conducted two laboratory experiments to investigate the relationships between Lucilia sericata (green bottle fly) and Calliphora vicina (blue bottle fly), expirated bloodstains, and pooled bloodstains on a range of surfaces (linoleum, wallpaper, textured paint). C. vicina and L. sericata changed bloodstain pattern morphology through feeding and defecation. They also deposited artifacts in rooms where blood was not present originally. Chemical presumptive tests (Hemastix^®, phenolphthalein, leucocrystal violet, fluorescein) were not able to differentiate between insect artifacts and bloodstains. Thus, C. vicina and L. sericata can confound bloodstain pattern analysis, crime scene investigation, and reconstruction. Crime scene investigators should be aware of these fundamental behaviors, and the effects that blow flies can have on expirated and pooled bloodstain patterns.     

Preliminary Observations on the Ability of Hyperspectral Imaging to Provide Detection and Visualization of Bloodstain Patterns on Black Fabrics

Abstract: The analysis of bloodstain patterns can assist investigators in understanding the circumstances surrounding a violent crime. Bloodstains are routinely subjected to pattern analysis, which is inherently dependent upon the ability of the examiner to locate and visualize bloodstain patterns on items of evidence. Often, the ability to properly visualize bloodstain patterns is challenging, especially when the stain patterns occur on dark and/or patterned substrates. In this study, preliminary research was performed to better understand how near‐infrared reflectance hyperspectral imaging (HSI) could be used to observe bloodstain patterns on commonly encountered black fabrics. The ability of HSI to visualize latent bloodstains on several commonly encountered substrates is demonstrated. The images acquired through HSI are of sufficient quality to allow for differentiation between stains produced from an impact mechanism or a transfer mechanism. This study also serves as a proof of concept in the differentiation of multiple staining materials. Because of its ability to generate spectral data, the data provide a preliminary separation of stains where more than one type of stain existed.     

Impinging blood droplets on different wettable surfaces: Impact phenomena,contact line motion,post-impact oscillation and dried stains

Understanding the underlying hydrodynamics of impinging blood droplets and finding out the physical parameters determining the bloodstain characteristics are of great importance in blood related forensic investigations. In this work, the impact of non-Newtonian blood droplets on solid surfaces ranging from lyophilic to superlyophobic was systematically investigated and compared to that of Newtonian droplets with a similar dynamic shear viscosity. We show that impinging blood droplets behave as low-viscosity Newtonian droplets in the short-time spreading, which is dominated by capillary and inertial forces, but their non-Newtonian viscoelasticity would notably affect the droplet retraction and post-impact oscillation occurring in large timescales. Whereas the strong liquid–solid adhesion and the non-Newtonian elongational viscosity hinder droplet recoiling and thus alter the impact phenomena on lyophobic and superlyophobic surfaces, the shear and elongational viscosities are coupled to result in higher damping coefficients of oscillating blood droplets after deposition, in comparison to that of impinging Newtonian droplets. The size of the dried bloodstain was found to be different from both the maximum spreading radius of the droplet that can reach during impact and the final radius of the deposited droplet after oscillation, and their correlations are highly dependent on the impact velocity and surface wettability. Moreover, the morphologic characteristics of the bloodstains would also be changed by varying either the impact velocity or the surface wettability. We envision that these findings can not only find applications in the bloodstain pattern analysis, but also provide useful information for medical diagnosis based on blood droplet test.     

The Reliability of Pattern Classification in Bloodstain Pattern Analysis,Part 1: Bloodstain Patterns on Rigid Non‐absorbent Surfaces

This study was designed to produce the first baseline measure of reliability in bloodstain pattern classification. A panel of experienced bloodstain pattern analysts examined over 400 spatter patterns on three rigid non‐absorbent surfaces. The patterns varied in spatter type and extent. A case summary accompanied each pattern that either contained neutral information, information to suggest the correct pattern (i.e., was positively biasing), or information to suggest an incorrect pattern (i.e., was negatively biasing). Across the variables under examination, 13% of classifications were erroneous. Generally speaking, where the pattern was more difficult to recognize (e.g., limited staining extent or a patterned substrate), analysts became more conservative in their judgment, opting to be inconclusive. Incorrect classifications increased as a function of the negatively biasing contextual information. The implications of the findings for practice are discussed.     

An Impact Velocity Device Design for Blood Spatter Pattern Generation with Considerations for High‐Speed Video Analysis,

A mechanical device that uses gravitational and spring compression forces to create spatter patterns of known impact velocities is presented and discussed. The custom‐made device uses either two or four springs (k₁ = 267.8 N/m, k₂ = 535.5 N/m) in parallel to create seventeen reproducible impact velocities between 2.1 and 4.0 m/s. The impactor is held at several known spring extensions using an electromagnet. Trigger inputs to the high‐speed video camera allow the user to control the magnet's release while capturing video footage simultaneously. A polycarbonate base is used to allow for simultaneous monitoring of the side and bottom views of the impact event. Twenty‐four patterns were created across the impact velocity range and analyzed using HemoSpat. Area of origin estimations fell within an acceptable range (ΔX_av = ?5.5 ± 1.9 cm, ΔY_av = ?2.6 ± 2.8 cm, ΔZ_av = +5.5 ± 3.8 cm), supporting distribution analysis for the use in research or bloodstain pattern training. This work provides a framework for those interested in developing a robust impact device.     

Differentiation of Twenty‐One Glitter Lip‐Glosses by Pyrolysis Gas Chromatography/Mass Spectroscopy*

Abstract: Differentiation of 21 glitter lip‐glosses from seven manufacturers was attempted by pyrolysis gas chromatography/mass spectroscopy. Samples were pyrolyzed on a ribbon probe at 800°C for 20 sec and analyzed with an Agilent^® 6890N Network GC System and Agilent^® 5973 Network Mass Selective Detector with MSD Productivity ChemStation^® Data Analysis software. The total ion chromatograms obtained were examined and differences in the presence or absence of certain chromatographic peaks corresponding to certain pyrolysis products (e.g., styrene, cyclohexane) noted. In cases where the total ion chromatograms between lip‐glosses were similar, select ion profiling was performed. Of the 21 lip‐glosses, 15 were differentiated by either the total ion chromatograms alone or through select ion profiling. Considering that lip‐glosses are typically worn by young women (who are disproportionately victims of sexual assault), this study offers the potential of being able to provide investigative leads in sexual assault investigations with evidentiary samples of this kind.     

Automatic Classification of Bloodstain Patterns Caused by Gunshot and Blunt Impact at Various Distances

The forensics discipline of bloodstain pattern analysis plays an important role in crime scene analysis and reconstruction. One reconstruction question is whether the blood has been spattered via gunshot or blunt impact such as beating or stabbing. This paper proposes an automated framework to classify bloodstain spatter patterns generated under controlled conditions into either gunshot or blunt impact classes. Classification is performed using machine learning. The study is performed with 94 blood spatter patterns which are available as public data sets, designs a set of features with possible relevance to classification, and uses the random forests method to rank the most useful features and perform classification. The study shows that classification accuracy decreases with the increasing distance between the target surface collecting the stains and the blood source. Based on the data set used in this study, the model achieves 99% accuracy in classifying spatter patterns at distances of 30 cm, 93% accuracy at distances of 60 cm, and 86% accuracy at distances of 120 cm. Results with 10 additional backspatter patterns also show that the presence of muzzle gases can reduce classification accuracy.     

Exsanguinated blood volume estimation using fractal analysis of digital images

The estimation of bloodstain volume using fractal analysis of digital images of passive blood stains is presented. Binary digital photos of bloodstains of known volumes (ranging from 1 to 7 mL), dispersed in a defined area, were subjected to image analysis using FracLac V. 2.0 for ImageJ. The box-counting method was used to generate a fractal dimension for each trial. A positive correlation between the generated fractal number and the volume of blood was found (R(2) = 0.99). Regression equations were produced to estimate the volume of blood in blind trials. An error rate ranging from 78% for 1 mL to 7% for 6 mL demonstrated that as the volume increases so does the accuracy of the volume estimation. This method used in the preliminary study proved that bloodstain patterns may be deconstructed into mathematical parameters, thus removing the subjective element inherent in other methods of volume estimation.     

Chemical Enhancement Techniques of Bloodstain Patterns and DNA Recovery After Fire Exposure*

Abstract: It is common in forensic casework to encounter situations where the suspect has set a fire to cover up or destroy possible evidence. While bloodstain pattern interpretation, chemical enhancement of blood, and recovery of deoxyribonucleic acid (DNA) from bloodstains is well documented in the literature, very little information is known about the effects of heat or fire on these types of examinations. In this study, a variety of known types of bloodstain patterns were created in a four‐room structure containing typical household objects and furnishings. The structure was allowed to burn to flashover and then it was extinguished by firefighters using water. Once the structure cooled over night, the interior was examined using a bright light. The bloodstains were evaluated to see if the heat or fire had caused any changes to the patterns that would inhibit interpretation. Bloodstain patterns remained visible and intact inside the structure and on furnishings unless the surface that held the blood was totally burned away. Additionally, a variety of chemical techniques were utilized to better visualize the patterns and determine the possible presence of blood after the fire. The soot from the fire formed a physical barrier that initially interfered with chemical enhancement of blood. However, when the soot was removed using water or alcohol, the chemicals used, fluorescein, luminol, Bluestar^®, and Hemastix^®, performed adequately in most of the tests. Prior to DNA testing, the combined phenolphthalein/tetramethyl benzidine presumptive test for the presence of blood was conducted in the laboratory on samples recovered from the structure in an effort to assess the effectiveness of using this type of testing as a screening tool. Test results demonstrated that reliance on obtaining a positive presumptive result for blood before proceeding with DNA testing could result in the failure to obtain useful typing results. Finally, two DNA recovery methods (swabbing the stain plus cutting or scraping the stain) were attempted to evaluate their performance in recovering samples in an arson investigation. Recovery of DNA was more successful in some instances with the swabbing method, and in other instances with the cutting/scraping method. Therefore, it is recommended that both methods be used. For the most part, the recovered DNA seemed to be unaffected by the heat, until the temperature was 800°C or greater. At this temperature, no DNA profiles were obtained.     

A Y‐Chromosomal Haplotype with Two Short Tandem Repeat Mutations*

Abstract:  The male‐specific Y‐chromosomal short tandem repeat (STR) is a useful tool in forensic casework. The Y haplotype comprised of 16 loci, which is amplified simultaneously by AmpFlSTR^® Yfiler^TM PCR kit and provides strong exculpatory evidence in individual identification. We reported a rare Y‐STR profile with a null allele at the DYS448 locus and an off‐ladder allele at the DYS456 locus, when genotyping material from a vaginal swab in an alleged rape case. Sequence analysis revealed that the DYS448 null allele was a true type of null allele because of a total deletion of 11 upstream repeats and 9 bp of the N₄₂ region, and there were numerous primer binding site mutations as well. The amplicon of the DYS456 locus was a small 92‐bp fragment that was off‐ladder, and sequencing analysis showed that there were only 10 repeats (AGAT)₁₀. This Y chromosome haplotype that was comprised of two variations provided helpful evidence for personal identification.     

Enhancing the Visibility of Injuries with Narrow‐Banded Beams of Light within the Visible Light Spectrum

The use of narrow‐banded visible light sources in improving the visibility of injuries has been hardly investigated, and studies examining the extent of this improvement are lacking. In this study, narrow‐banded beams of light within the visible light spectrum were used to explore their ability in improving the visibility of external injuries. The beams of light were induced by four crime‐lites^® providing narrow‐banded beams of light between 400 and 550 nm. The visibility of the injuries was assessed through specific long‐pass filters supplied with the set of crime‐lites^®. Forty‐three percent of the examined injuries improved in visibility by using the narrow‐banded visible light. In addition, injuries were visualized that were not visible or just barely visible to the naked eye. The improvements in visibility were particularly marked with the use of crime‐lites^® “violet” and “blue” covering the spectrum between 400–430 and 430–470 nm. The simple noninvasive method showed a great potential contribution in injury examination.     

Bloodstain pattern analysis--casework experience

The morphology of bloodstain distribution patterns at the crime scene carries vital information for a reconstruction of the events. Contrary to experimental work, case reports where the reconstruction has been verified have rarely been published. This is the reason why a series of four illustrative cases is presented where bloodstain pattern analysis at the crime scene made a reconstruction of the events possible and where this reconstruction was later verified by a confession of the offender. The cases include various types of bloodstains such as contact and smear stains, drop stains, arterial blood spatter and splash stains from both impact and cast-off pattern. Problems frequently encountered in practical casework are addressed, such as unfavourable environmental conditions or combinations of different bloodstain patterns. It is also demonstrated that the analysis of bloodstain morphology can support individualisation of stains by directing the selection of a limited number of stains from a complex pattern for DNA analysis. The complexity of real situations suggests a step-by-step approach starting with a comprehensive view of the overall picture. This is followed by a differentiation and analysis of single bloodstain patterns and a search for informative details. It is ideal when the expert inspecting the crime scene has also performed the autopsy, but he definitely must have detailed knowledge of the injuries of the deceased/injured and of the possible mechanisms of production.     

Statistical Evaluation of Alternative Light Sources for Bloodstain Photography

Bloodstain photography is important in forensic applications, especially for bloodstain pattern analysis. This study compares the enhancement effect of bloodstain photography using three different types of light source: fluorescent white light, near‐ultraviolet (UV) light‐emitting diode (LED) light, and 410 nm LED light. Randomized complete block designs were implemented to identify the lighting that would statistically produce the best enhancement results for bloodstains on different types of surfaces. Bloodstain samples were prepared on white cotton, brown carpet, tar road, and wood. These samples were photographed in darkroom conditions using a Canon EOS 50D digital SLR camera, with Canon EFS 60 mm f/2.8 Macro USM lens. Two‐way analysis of variance and Fisher's least significant difference test were used to analyze the contrast of the images. The statistical analysis showed that 410 nm light is the best among the tested lights for enhancing bloodstains on the tested surfaces, where the contrast of bloodstain to background was the highest.     

Improved Area of Origin Estimation for Bloodstain Pattern Analysis Using 3D Scanning

An important component of crime scene reconstruction is bloodstain pattern analysis (BPA). Where BPA concerns impact patterns, estimating the area of origin is critical information for scene reconstruction. Traditionally, this is achieved by measuring individual bloodstains and performing trigonometric calculations; however, 3D scanning has been proposed as a viable alternative for overcoming logistical and practical concerns with the manual method. Therefore, this project aimed to establish whether the FARO Focus 3D scanner and FARO Zone 3D software can improve the accuracy of area of origin estimates relative to the manual method. We created a series of eight bloodstain impact patterns and performed paired analysis using the two methods to estimate areas of origin for each pattern. Our data suggested that FARO-derived estimates were generally more accurate than using the manual method. FARO-estimated heights of origin areas were generally closer to the true distance. Both methods underestimated the distance from the wall for most patterns originating 150mm or greater from the wall, but overestimated distances for patterns originating closer to the wall. The degree to which distances were underestimated increased significantly the further the blood source was from the wall and was greater for FARO-derived estimates. The results of this research contribute to the validation of these instruments for operational implementation for BPA and should be considered alongside the practical benefits of 3D scanning relative to manual methods. Further, 3D scanning can provide reliable BPA reconstruction documentation for technical review and court presentation.     

Improving the point of origin determination in bloodstain pattern analysis

In bloodstain pattern analysis, it is important to know the point of origin (PO) of an impact pattern. This point can be estimated by means of the stringing method, the tangent method, or by commercially available computer programs. In this study, the accuracy of two computer programs was investigated under different conditions. Impact patterns were created by means of a modified mouse trap, and subsequently the PO was calculated. By examining the characteristics of single bloodstains, the influence on the deviation could be determined. To improve the estimation of the PO, it is important to select bloodstains that lie close to the presumable location of the blood source, that are large (width >1.5 mm) and that show an elliptical form. If possible, bloodstains from different walls should be taken into account. Our recommendations may improve the PO determination of impact patterns.