Fundamental studies of bloodstain formation and characteristics

A detailed understanding of blood droplet impact dynamics and stain formation is an essential prerequisite to the interpretation of both individual bloodstains and spatter patterns. The current literature on theoretical models for the spreading and splashing of liquid drops on surfaces relevant to the forensic context of bloodstain formation has been reviewed. These models have been evaluated for a paper substrate using experimental data obtained as function of droplet size, impact velocity and angle. It is shown that for perpendicular impact there are fairly simple mathematical models for the spreading diameter and the number of scallops or spines formed around the stain though these have quite limited ranges of validity in their basic form. In particular, predictions for the diameter are best for small droplets impacting at high velocity and the number of spines saturates for higher impact velocities. In the case of spreading, a modification to the energy conservation model is found to provide excellent agreement with experimental stain diameters across a wide range of impact velocities. For non-perpendicular impact, the width of stains is found to depend principally on the normal component of impact velocity and may be predicted by an appropriate modification to the expression for the perpendicular case. Limitations in the calculation of impact angle from the stain aspect ratio are identified and a theoretical basis for the prediction of spines around an elliptical stain is proposed. Some key issues for future research are identified which include a systematic, quantitative study of the effect of surface properties on bloodstain formation.     

The influence of fabric surface characteristics on satellite bloodstain morphology

Bloodstains on fabrics such as clothing, soft furnishings or carpets are often encountered in casework. These stains often have a distinctive morphology that includes satellite stains, thought to be a highly sensitive feature that is a function of surface roughness. This study presents the findings of experimental studies conducted with proxy blood on two fabrics, similar in labeled composition, to assess the influence of fabric type on satellite stain generation. The morphology of proxy blood stains on the two fabric types were found to be statistically distinguishable from one another, with the volume of satellite stains generated being dependent upon the surface roughness of the fabric. These findings provide an initial step that illustrates the viability of providing an empirical evidence base for the interpretation of satellite stains in forensic blood pattern analysis (BPA).     

The utility of polyester and cotton as swabbing substrates for the removal of cellular material from surfaces

Various types of cotton and polyester fabrics were tested to ascertain the optimal physical and chemical characteristics of fabrics needed for the removal of cellular material from surfaces. DNA quantitation values obtained on dried saliva stains showed no difference between cotton and polyester across all constructions and solvent conditions. Fabrics used dry and with water yielded higher quantitation values than those used with isopropanol. Quantitation values were also higher for wovens and nonwovens than knits across all solvent conditions. Low thread count fabrics used with water yielded higher quantitation values, but no correlation between thread count and quantitation values was observed with dry fabrics. A low thread count woven fabric, however, outperformed other tested fabrics when swabbing object surfaces in a highly used room. Full DNA profiles from fingerprints on glass surfaces were obtained with low thread count woven and nonwoven fabrics but not with the knit fabric tested.     

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.     

Acid phosphatase screening — Wetting test paper or wetting fabric and test paper?

Detecting and locating semen stains is addressed by identifying the associated acid phosphatase activity of semen. The recovery of semen stains is critical as it can, via DNA testing, address the possible source(s) of the semen and may aid in the interpretation of a case. The purpose of these experiments, carried out on behalf of the Body Fluids Forum, was to consider whether wetting the test paper alone or wetting the semen stained fabric and the test paper affected the detection and location of the semen stains on various fabric types, or the subsequent recovery of spermatozoa from these fabrics. It became evident that the preferred approach varied depending on the fabric type being tested but that more often than not, wetting both the fabric and the test paper had a detrimental effect on the recovery of spermatozoa.     

被洗织物上潜血痕迹发现方法的比较

目的探索被洗涤后衣物上潜血痕迹的发现方法。方法用联苯胺法、鲁米诺法和新型仪器TL-445激光生物检材发现仪寻找不同织物上不同洗涤强度的潜在血迹。结果联苯胺仅能发现部分纯棉织物上潜血,鲁米诺能显示全部纯棉织物上血迹,TL-445激光生物检材发现仪能显示所有纯棉、纯化纤织物上血迹。结论用TL-445激光生物检材发现仪检测相比联苯胺法和鲁米诺法更适合洗涤后织物上潜血痕迹的发现。     

PCR-based detection of salivary bacteria as a marker of expirated blood

Distinguishing between bloodstains caused by a spatter pattern or by expirated blood may be crucial to a forensic investigation. Expirated blood is likely to be contaminated with saliva but current techniques have limited sensitivity, especially with small bloodstains. We report that a PCR assay, designed to detect salivary bacteria, can amplify streptococcal DNA from saliva stains applied to fabrics for at least 62 days after seeding. Bacterial DNA was detected when 0.01 µl of saliva was present in the stain and the amplification was not affected by contamination with blood. These findings indicate that PCR amplification of salivary microbial DNA may have application in the identification of expirated bloodstains in forensic case-work.     

Improving the effectiveness of fluorescence for the detection of semen stains on fabrics

The paper describes the use of the Polilight, a light source based on a xenon arc lamp, to exploit the fluorescence properties of semen as an aid to searching fabrics for stains in sexual assault cases. The broad excitation spectrum of semen allows the fluorescence to be generated at a range of wavelengths. This permits the excitation and emission conditions to be selected that minimize interference from background fluorescence of the fabric and thereby optimizes the contrast between the fabric and the stain. A common method for the observation of fluorescence is the use of colored plastic goggles or filters. The paper shows that the detection of fluorescence from semen stains is significantly enhanced using appropriate interference filters.     

Morphology of low-velocity impact stains produced from single drops of blood

Systematic variation of blood droplet volume, the distance fallen and the surface (paper, wood, plastics, tiles) led to the conclusion that the size and the shape of the stains ("fingers", satellites) allowed to deduce the distance fallen but only if the actual surface structure was known. We found that detailed photography at the crime scene was necessary, yet experiments have to be performed due to the extreme influence of the actual surface texture on all characteristics (size, spines, peripheral spatter) of the blood stains.     

Chemical enhancement of footwear impressions in blood on fabric – Part 1: Protein stains

A range of protein stains were utilised for the enhancement of footwear impressions on a variety of fabric types of different colours with blood as a contaminant. A semi-automated stamping device was used to deliver test impressions at a set force to minimise the variability between impressions; multiple impressions were produced and enhanced by each reagent to determine the repeatability of the enhancement. Results indicated that while most protein stains used in this study successfully enhanced impressions in blood on light coloured fabrics, background staining caused interference on natural fabrics. Enhancement on dark coloured fabrics was only achieved using fluorescent protein stains, as non-fluorescent protein stains provided poor contrast.A further comparison was performed with commercially available protein staining solutions and solutions prepared within the laboratory from the appropriate chemicals. Both solutions performed equally well, though it is recommended to use freshly prepared solutions whenever possible.     

Predicting the position of the source of blood stains for angled impacts

Droplets of pig's blood were dropped onto paper at different angles to the horizontal to produce blood stains. Impact velocities varied from 1.82 to 5.76 m/sec, drop size from 3.7 to 5.0 mm in diameter, and the surface sloped at angles between 22.7 degrees and 90 degrees to the horizontal. From the data a single equation relating stain size to drop size and velocity for all impact angles was produced; ab = 111.74 (Re(1/2)We(1/4))(0.75)D(o)D(o) + 0.00084 with R(2) = 0.88, where a is the stain width, b the stain length, Re the Reynolds number, and We the Weber number. A second equation related the number of spines, N, to drop size, velocity, and surface slope for all impact angles as N = 0.76 We(0.5) sin(3)theta with R(2) = 0.9, where theta is the impact angle. Combining these equations the impact velocity can be determined and hence the position of the stain's source.     

Analysis of Luminescent Gunshot Residue (LGSR) on Different Types of Fabrics

The collection of gunshot residue on fabric can be an arduous task due to the microscopic size of particles (blind collection) and sheddability of some fabrics. The introduction of luminescent markers and consequent formation of luminescent gunshot residue (LGSR) can facilitate this analysis. In this study, different fabrics were analyzed in order to verify the persistence of the LGSR on them, the possibility of collecting and analyzing particles by video spectral comparator (VSC) and SEM/EDS. Also, different colored fabrics were used as targets in order to investigate influence of fabric color on LGSR visualization. Furthermore, the influence of the fabric type in the distribution of the LGSR deposited around the projectile´s hole entrance was evaluated. The fabric sheddability did not alter collection of the particles or analysis. It was possible to observe and collect LGSR on all tested fabrics, even after the fabric had been shaken, or in colored fabrics.     

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.     

Deducing drop size and impact velocity from circular bloodstains

An experimental study was done to determine the diameter and velocity of blood drops falling on a surface by measuring the size of bloodstains they produced and counting the number of radial spines projecting from them. Bloodstains were formed by releasing drops of pig blood with a range of diameters (3.0-4.3 mm) and impact velocities (2.4-4.9 m/s), onto four different flat surfaces (glass, steel, plastic, paper) with varying roughness (0.03-2.9 microm). High-speed photography was used to record drop impact dynamics. Bloodstain diameters and the number of spines formed around the rim of stains increased with impact velocity and drop diameter. Increasing surface roughness reduced stain diameter and promoted merging of spines, diminishing their number. Equations are presented that explicitly relate drop diameter and impact velocity to measurements of stain diameter and number of spines.     

Distinction of bloodstain patterns from fly artifacts

Forensic scientists may encounter blood spatter at a scene which may be pure or a mixture of fly artifacts and human bloodstains. It is important to be able to make an informed identification, or at least advanced documentation of such stains since the mechanics of production of fly artifacts are not determinable to the crime scene reconstructionist from regular police forces. We describe three cases in which experiments and crime scene reconstruction led to additional information. Case 1: Above the position of a victim, numerous blood stains of the low-high-velocity type were found. Exclusion of these stains being caused by force (but instead caused by the activity of adult blow flies) by use of the following observations that were confirmed in experiments: (a) sperm-/tadpole-like structure with length > width, (b) random directionality, and (c) mixture of round symmetrical and teardrop shaped stains. Case 2: A reddish spatter field was found on a fan chain two rooms away from the place where a dead woman was found. Localization of the spatter on the bottom end of the surface hinted strongly towards fly activity. Case 3: Double homicide; submillimeter stains were found on a lamp between the two corpses. Activity of flies was less likely compared to alternative scenario of moving lampshade and violent stabbing.     

The use of Polilight in the detection of seminal fluid, saliva, and bloodstains and comparison with conventional chemical-based screening tests

Biological stains can be difficult to detect at crime scenes or on items recovered from crime scenes. The use of a versatile light source may assist in their detection. The ability of Polilight to locate potential semen, saliva, and blood stains on a range of substrates and at different dilutions was tested. We also tested the use of Polilight in comparison with conventional chemical-based presumptive screening tests such as acid phosphatase (AP), Phadebas, and luminol, often used in casework for detecting potential semen, saliva, and blood stains, respectively. The Polilight was able to locate stains that were not apparent to the naked eye. The color of the material on which a stain is deposited can have an effect on the detectibility of the stain. The Polilight was found to be comparable with the AP and Phadebas tests in terms of its sensitivity. In a comparative study between the AP test and Polilight on 40 casework exhibits, one false-negative result was observed when using the Polilight. On a series of mock casework exhibits it was determined that the Polilight can be used successfully to locate saliva stains for DNA analysis. The sensitivity of luminol for detecting potential bloodstains was greater than that of Polilight; however the Polilight has particular application in instances where a bloodstain may have been concealed with paint. Overall, the Polilight is a relatively safe, simple, noninvasive, and nondestructive technique suitable for use in forensic casework.     

Detection and visualization of human tears using alternate light sources for forensic purposes

The current scientific techniques for locating body fluids focus on quick and effective methodologies for easy and reliable identification. Efficient detection and identification of body fluids play a vital role in establishing the ‘corpus delecti’ of a crime. Non-destructive techniques such as the use of Alternate Light Sources (ALS) have been exploited for crime scene searches over large areas and detection of body fluids such as blood, semen, vaginal secretions, and saliva on a range of substrates. Tears are rarely found but can be considered as potential body fluid evidence due to their unique biochemical and molecular properties. Tears are secreted in response to physical or emotional stimuli. Due to the small volume of secretions, they are often overlooked in the crime scene. Tears may be found on surfaces such as clothing, bedding, tissue, handkerchief, or balaclava. The use of ALS to locate tears on tissue paper and fabric surfaces was tested which were not apparent to the naked eye. Tears stains were successfully detected on surfaces of forensic interest with varying sample ages up to three months with a broad excitation spectrum between 254 nm and 410 nm. Dried stains on tissue paper and fabric substrates were better detected with sharp margins, clear stain pattern visibility, and fluorescence intensity in comparison with moist and fresh stains. Tears stains can hence be detected with the use of ALS and suitable filter combinations under normal conditions and do not require any specific settings to locate them. These findings are suggestive for easy and quick identification of tears on large surfaces and as a presumptive test for forensic casework evidence examination.     

Changes in the morphology and presumptive chemistry of impact and pooled bloodstain patterns by Lucilia sericata (Meigen) (Diptera: Calliphoridae)

Bloodstain pattern analysis can be critical to accurate crime scene reconstruction. However, bloodstain patterns can be altered in the presence of insects and can confound crime scene reconstruction. To address this problem, we conducted a series of controlled laboratory experiments to investigate the effect of Lucilia sericata (Meigen) on impact bloodstains and pooled bloodstains in association with three combinations of common surfaces (linoleum/painted drywall, wood floor/wallpaper, and carpet/wood paneling). L. sericata fed from the pooled bloodstains and added insect stains through regurgitation and defecation of consumed blood. L. sericata formed defecatory trails of insect stains that indicated directionality. Defecatory stains fluoresced when viewed at 465 nm with an orange filter. These observations differed from Calliphora vicina insect stains because feeding on blood spatter was not observed and trails of insect stains were formed by L. sericata. The fluorescence of defecatory stains can be used as a method to detect insect stains and discriminate them from real bloodstains.     

Chemical enhancement of footwear impressions in blood on fabric — Part 3: Amino acid staining

Enhancement of footwear impressions, using ninhydrin or ninhydrin analogues is not considered common practice and such techniques are generally used to target amino acids present in fingermarks where the reaction gives rise to colour and possibly fluorescence. Ninhydrin and two of its analogues were used for the enhancement of footwear impressions in blood on various types, colours and porosities of fabric. Test footwear impressions on fabric were prepared using a specifically built rig to minimise the variability between each impression. Ninhydrin enhancement of footwear impressions in blood on light coloured fabric yielded good enhancement results, however the contrast was weak or non-existent on dark coloured fabrics. Other ninhydrin analogues which have the advantage of fluorescence failed to enhance the impressions in blood on all fabrics. The sequential treatment of impressions in blood on fabric with other blood enhancing reagents (e.g. protein stains and heme reagents) was also investigated.