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.     

DNA investigative lead development from blood and saliva samples in less than two hours using the RapidHIT™ Human DNA Identification System

The RapidHIT™ Human DNA Identification System produces full STR DNA profiles (sample in, results out) in 90 min. Here we present results from a new protocol on the RapidHIT System, designed for crime scene samples. Data are presented for blood and saliva samples, with results for a range of samples and substrates. Success rates and sensitivity display a high level of performance for the sample types evaluated in the study and demonstrate the efficacy of the new protocol for crime scene samples.     

A Novel Solvent-based Method to Separate Duct Tape from Porous Surface for Fingerprint Development

In this study, we have proposed a novel solvent-based method using a specific concentration of 3M™ Novec™ HFE-72DE and 7200 solvents to untangle duct tape from porous article with minimal impact to the quality of latent fingerprint deposited on the sticky-side surface of duct tape. A series of experiments determined that a mixture of 30% HFE-72DE:7200 (v/v) was found to be the most effective to separate various brands of duct tape from different types of porous surface, including office copy paper, newspaper, cardboard, and tissue wipe, which had been stored for up to 30 days before untangling. Further studies also revealed that 30% HFE-72DE:7200 was compatible with three common fingerprint development methods for porous articles, namely ninhydrin, indanedione-zinc, and physical developer. The nonflammability and low toxicity nature of this novel solvent mixture also make it ideal for separating duct tape from porous surface in laboratory or at crime scene.     

Evaluating the use of hypoxia sensitive markers for body fluid stain age prediction

To augment DNA profiling and body fluid identification techniques efforts are being made to increase the amount of information available from a crime scene stain, which includes efforts to identify externally visible characteristics through phenotypic analysis. A key question surrounding crime scene stains is the length of time between deposition of the stain and its subsequent recovery, in that is the stain recovered related to the incident in question or from a previously deposited stain number of weeks earlier? The inability to answer this fundamental question has a detrimental effect upon the successful completion of a criminal investigation. Once a body fluid leaves the body, the oxygen concentration in the environment changes; therefore, it may be that this change could cause a change in the expression of hypoxia-sensitive biomarkers. Here, a range of bloodstains, liquid saliva and liquid semen samples were collected at 0 days, 7 days, 14 days, 21 days and 28 days of degrading at room temperature (19–22 °C), before undergoing total RNA extraction and cDNA synthesis. Blood was recovered from filter paper with 3 mm², with saliva and semen being left in their tubes and swabbed at the appropriate times. All samples then underwent quantitative PCR targeting Vascular Endothelial Growth Factor A (VEGFA) and Hypoxia-Inducible Factor 1 Alpha (HIF1A), with B-Actin (ACTB) as a reference gene. A range of linear and quadratic correlation values was obtained from the qPCR data and used to develop a predictive model with a mean absolute deviation (MAD) of 4.2, 2.1, and 5 days for blood, saliva, and semen respectively. Blind testing indicated that a stain age prediction model based upon VEGFA with ACTB as a reference gene could be used on samples up to four weeks old with a margin of error ranging from 2 days through to 5 days. While a sizeable potential time frame exists using this model; this represents a significant step towards the target of having an accurate stain age prediction model.     

Assessment of body fluid identification and DNA profiling after exposure to tropical weather conditions

Despite current advances in body fluid identification, there are few studies evaluating the effect of environmental conditions. The present work assessed the detection of body fluids, blood, semen, and saliva, through lateral flow immunochromatographic (LFI) tests, exposed to tropical weather conditions over time, also evaluating the possibility of obtaining STR (short tandem repeat) profiles and identifying mitochondrial DNA (mtDNA) polymorphisms. Blood, semen, saliva samples, and mixtures of these fluids were deposited on polyester clothes and exposed to open-air tropical weather conditions for 1 month. The test versions from LFI (SERATEC®, Germany) Lab and crime scene (CS) used for the detection – one per each body fluid type – demonstrated that it is possible to identify body fluids and their mixtures up to 14 days after deposition. At 30 days, blood and semen were detected but not saliva. Full STR profiles were obtained from 14-day-old blood samples, and partial profiles were obtained from the remaining samples. It was possible to sequence mtDNA in the samples previously analyzed for STR profiling, and haplogroups could be assigned. In conclusion, this study demonstrated for the first time the possibility of body fluid identification and DNA profiling after exposure to tropical weather conditions for 1 month and also demonstrated the value of mtDNA analysis for compromised biological evidence.     

Highly Sensitive Detection of Blood by Surface Enhanced Raman Scattering,,

Raman spectroscopy for forensic body fluid analysis has received some attention due to the nondestructive nature and potential application for identification at the crime scene; however, its usage has been limited by low detection sensitivity. Surface enhanced Raman scattering (SERS) was evaluated for blood identification for forensic applications. Specifically, a SERS‐active substrate was fabricated, composed of nickel nanotips coated with Ag nanoparticles. Compared with a conventional substrate, the SERS substrate enhanced Raman scattering by more than two orders of magnitude and allowed blood to be identified to a dilution of 1:100,000. Blood was also successfully detected by swabbing the SERS substrate directly on mock evidence. Most importantly, Raman spectra obtained by swabbing the SERS substrate on blood stains were free of luminescence even when blood was deposited on luminescent fabrics. The nondestructive character, simplicity of sample preparation, and high sensitivity make SERS a prime candidate for field and laboratory‐based blood identification.     

The Influence of New Technologies on the Visual Attention of CSIs Performing a Crime Scene Investigation

Currently, a series of promising new tools are under development that will enable crime scene investigators (CSIs) to analyze traces in situ during the crime scene investigation or enable them to detect blood and provide information on the age of blood. An experiment is conducted with thirty CSIs investigating a violent robbery at a mock crime scene to study the influence of such technologies on the perception and interpretation of traces during the first phase of the investigation. Results show that in their search for traces, CSIs are not directed by the availability of technologies, which is a reassuring finding. Qualitative findings suggest that CSIs are generally more focused on analyzing perpetrator traces than on reconstructing the event. A focus on perpetrator traces might become a risk when other crime‐related traces are overlooked, and when analyzed traces are in fact not crime‐related and in consequence lead to the identification of innocent suspects.     

The Effects of Substance Use on Offender Crime Scene Behavior

Abstract:  Substance use has an effect on an individual's propensity to commit acquisitive crime with recent studies showing substance users more likely to leave forensic material at a crime scene. An examination of acquisitive crime solved in Northamptonshire, U.K., during 2006 enabled 70 crime scene behavior characteristics to be analyzed for substance and nonsubstance use offenders. Logistical regression analyses have identified statistically significant crime scene behavior predictors that were found to be either present at or absent from the crime scene when the offender was a substance user. Most significant predictors present were indicative of a lack of preparation by the offender, irrational behavior, and a desire to steal high value, easily disposed of, property. Most significant predictors absent from the crime scene were indicative of more planning, preparation, and execution by the offender. Consideration is given to how this crime scene behavior might be used by police investigators to identify offenders.     

"Biological identikit": Development of a SNPs-panel for the analysis of forensic DNA phenotyping and ancestry

Personal identification in mass disasters and in crimes is essential for humanitarian, ethical and legal reasons. In these contexts, when individuals cannot be identified by standard forensic DNA analysis, the Forensic DNA Phenotyping and the analysis of the biogeographical ancestry could help. The aim of this study was to evaluate the potential of a new panel of 891 SNPs in predicting phenotypic traits and biogeographical origin to create a “biological identikit”. In addition to fresh biological material, old evidence found at the crime scene or extracted and long-term stored DNA were tested with 41 SNPs for phenotyping and 850 SNPs for ancestry. All the SNPs were successfully incorporated into a single two-step multiplex PCR reaction using the IonAmpliSeq ™ Library Plus and applied for massive parallel sequencing with the Ion S5 platform using up to 0.05 ng/µL of DNA. The analysis of the results was carried out with an in-house predictive algorithm and consulting 20 population databases. By comparing the results obtained with identikit or video-photographic surveys, it was possible to predict phenotype and ancestry with an accuracy greater than 90%. While these new markers cannot identify a specific individual, they can be a valuable investigative tool.     

The Morphology of Fecal and Regurgitation Artifacts Deposited by the Blow Fly Lucilia cuprina Fed a Diet of Human Blood

Fly feces and regurgitation deposits may be mistaken for bloodstain patterns at a crime scene, potentially compromising event reconstruction and/or misdirecting police resources. In some instances, these artifacts contain sufficient human biological material to generate a full DNA profile, sometimes 2 years after deposition. Clearly, it is important that investigators can make the distinction between artifacts and bloodstains. This study examined 6645 artifacts deposited on a smooth, nonporous surface after Lucilia cuprina were fed human blood. Artifacts were also compared with bloodstains on a variety of other surfaces. Both similarities and differences were found between artifacts and bloodstains, highlighting the need for an identification system to assist personnel with little training in bloodstain pattern analysis. The morphology of the artifacts has been described so that these deposits may be more clearly distinguished from bloodstains, targeted by crime scene personnel as potential sources of human DNA, and/or identified as potential evidence contaminants. Flowcharts have been devised to facilitate the analysis.     

Palynological Investigation of Mummified Human Remains

Pollen analysis was applied to a mummified homicide victim in Nebraska, U.S.A., to determine the location of death. A control sample showed the normal ambient pollen in the garage crime scene. Ambient windborne types, common in the air of the region, dominated the control. Internal samples were analyzed from the sacrum, intestine, and diaphragm. Microfossils were recovered from the rehydrated intestine lumen. The intestinal sample was dominated by Brassica (broccoli). The sacrum sample was high in dietary types but with a showing of ambient types. The pollen from the diaphragm was dominated by ambient pollen similar to the control samples. The discovery of diverse pollen spectra from within a single mummy was unexpected. They show that ingested and inhaled pollen mixed in the corpse. The data linked the decedent to a specific crime scene in her Nebraska home in the southern tier of eastern counties on the border with Kansas.     

The use of error rates to predict and characterize the efficacy of presumptive field tests

From a forensic perspective, a presumptive test, one which indicates the presence or absence of a certain target material such as blood, is an invaluable tool. Among these tests, there are different specificities, sensitivities, and shelf lives. The accuracy of a test is an algebraic combination of the specificity and sensitivity of the test. Each test has limitations as given by its false positive and false negative rates. The aim of this study was to illustrate how the false positive and false negative rates are to be properly determined using a simulation study for the phenolphthalein test. New presumptive tests must be properly evaluated/validated through testing of commonly encountered household items and other potentially probative items usually found at crime scenes, however, the makeup of test sets must appropriately capture all error rates. In order to correctly use these results when the test is applied to an unknown sample recovered at a crime scene, the error rates cannot be applied directly to estimate whether or not the sample is actually the analyte of interest. In a validation study, the forensic scientist calculates the false positive rate as the p(Positive Reaction|Blood), whereas at the scene, the crime scene investigator wishes to determine the p(Blood|Positive Reaction). All crime scene investigators need to ensure that the conditional is not transposed when interpreting such results. Furthermore, this work provides a model for the assessment of a multiple test diagnostic system intended for investigators.     

Juvenile firesetters: Crime scene actions and offender characteristics

Purpose. To investigate whether and to what extent the thematic structure of crime scene actions in arsons identified in Canter and Fritzon (1998) is replicated for juvenile firesetters and to explore whether any associations between the crime scene action themes and offender characteristics would be evident. Methods. The crime scene actions and offender characteristics of 61 male and 5 female juvenile firesetters (aged 6‐17 years) were examined. The data were drawn from a larger database originally collected and content analysed in Fritzon (1998). In total, 43 dichotomous crime scene actions, 17 offender background characteristics and offender criminal record variables had been coded. Smallest space analysis was employed to examine the configuration of crime scene action and offender characteristic variables. The associations between the crime scene actions and offender characteristics, as well as the criminal record variables, were analysed using the correlation. Results. Distinct structural themes for crime scene actions were found in juvenile firesetting, similar to those identified in Canter and Fritzon (1998). Contrary to Canter and Fritzon, only two groups of background characteristics were identified, depressed and delinquent, the latter being more common and related to an instrumental form of firesetting. The expressive form of firesetting was associated with offenders' psychopathology and female gender. The presence of a crime scene action theme was associated with the offender's age. Conclusions. The structural themes of firesetting behaviour appear to transpire early. The background characteristics of juvenile firesetters indicate that juvenile firesetting is often associated with antisocial behaviour and psychopathology, deserving, therefore, disparate prevention, intervention and investigation programmes.     

Real-time forensic DNA analysis at a crime scene using a portable microchip analyzer

An integrated lab-on-a-chip system has been developed and successfully utilized for real-time forensic short tandem repeat (STR) analysis. The microdevice comprises a 160-nL polymerase chain reaction reactor with an on-chip heater and a temperature sensor for thermal cycling, microvalves for fluidic manipulation, a co-injector for sizing standard injection, and a 7-cm-long separation channel for capillary electrophoretic analysis. A 9-plex autosomal STR typing system consisting of amelogenin and eight combined DNA index system (CODIS) core STR loci has been constructed and optimized for this real-time human identification study. Reproducible STR profiles of control DNA samples are obtained in 2 h and 30 min with ≤0.8 bp allele typing accuracy. The minimal amount of DNA required for a complete DNA profile is 100 copies. To critically evaluate the capabilities of our portable microsystem as well as its compatibility with crime scene investigation processes, real-time STR analyses were carried out at a mock crime scene prepared by the Palm Beach County Sheriff's Office (PBSO). Blood stain sample collection, DNA extraction, and STR analyses on the portable microsystem were conducted in the field, and a successful “mock” CODIS hit was generated on the suspect's sample within 6 h. This demonstration of on-site STR analysis establishes the feasibility of real-time DNA typing to identify the contributor of probative biological evidence at a crime scene and for real-time human identification.     

犯罪现场生物检材的发现、提取策略

在改进现场生物检材的发现和提取技术的同时,也要进行理论创新和体系构建,才能够更好地指导实践。全局的观点、具体案件具体分析和犯罪现场重建对于现场勘查中有效地发现、提取和利用生物检材是十分重要的。     

Development of enhanced sensitivity protocols on the RapidHIT™ 200 with a view to processing casework material

The RapidHIT™ 200 device from IntegenX® provides a sample-to-profile platform that is capable of processing a variety of sample types. In this study we review the sensitivity of the ‘Run Other’ protocol for processing crime stain type samples containing various input quantities of DNA using the AmpFℓSTR® NGMSElect™ Express PCR Amplification Kit cartridges available from IntegenX®. The range of DNA inputs which achieved useable results were not as desired and therefore various enhancements to the instruments extraction processes were investigated. These studies showed an improvement in the range of DNA input templates that could by processed on the RapidHIT™ 200 by using the enhanced methods and resulted in three new run protocols.     

Pollen on grass clippings: putting the suspect at the scene of the crime.

In a case of alleged sexual assault, the pollen content of samples of grass clippings and soil from the suspect's clothing and shoes was compared to that of a sample of grass clippings from the alleged crime scene (a grassy area) to determine whether or not the suspect had been at the scene. The clothing and shoe samples showed a very strong correlation with each other and with the sample from the alleged crime scene in the combination of the different types of pollen present, very strongly supporting the contention that the suspect had been at the scene.     

Messenger RNA Profiling for Forensic Body Fluid Identification： Research and Applications

Identifying the origin of body fluids left at a crime scene can give a significant insight into crime scene reconstruction by supporting a link betw een sample donors and actual criminal acts. How ev-er, the conventional body fluid identification methods are prone to various limitations, such as time con-sumption, intensive labor, nonparallel manner, varying degrees of sensitivity and limited specificity. Re-cently, the analysis of cell-specific messenger RNA expression （mRNA profiling） has been proposed to supplant conventional methods for body fluid identification. Since 2011, the collaborative exercises have been organized by the European DNA Profiling Group （EDNAP ） in order to evaluate the robustness and reproducibility of mRNA profiling for body fluid identification. The major advantages of mRNA profil-ing, compared to the conventional methods, include higher sensitivity, greater specificity, the ability of detecting several body fluids in one multiplex reaction, and compatibilitywith current DNA extraction and analysis procedure. In the current review ,we provided an overview of the present know ledge and detection methodologies of mRNA profiling for forensic body fluid identification and discussed its possi-ble practical application to forensic casew ork.