Styryl dye coated metal oxide powders for the detection of latent fingermarks on non-porous surfaces

Conventional fingermark powders rely on contrast induced by absorption/reflection (e.g. black powder) or luminescence in the visible region (e.g. Blitz Green(?)). In most cases, these powders provide sufficient contrast; however, in some circumstances surface characteristics can interfere with the visualisation of powdered fingermarks. Visualisation in the near infra-red (NIR) region, however, has been shown to eliminate interferences commonly encountered in the visible region. In this study, a mixture of rhodamine 6G and the NIR laser dye styryl 11 (STaR 11) was coated onto an aluminium oxide nanopowder and then mixed with silver magnetic powder to develop and visualise fingermarks in the NIR. When compared to Blitz Green(?), it was determined that the STaR 11 magnetic powder was better suited for marks deposited on textured surfaces and for older marks, whereas Blitz Green(?) performed better on smooth glossy surfaces. The ability of the STaR 11 mixed dye formulation to be visualised in both the visible and NIR regions also provides a significant advantage over conventional luminescent fingermark powders.     

Identification and age estimation of blood stains on colored backgrounds by near infrared spectroscopy

Non-destructive identification and subsequent age estimation of blood stains are significant steps in forensic casework. The latter can provide important information on the temporal aspects of a crime. As previously shown, visible spectroscopy of blood stains on white backgrounds can successfully be used for their identification and age estimation. The use of this technique however, is hampered by dark backgrounds. In the present study the feasibility to use near infrared (NIR) spectroscopy was evaluated for blood stain identification and age estimation on dark backgrounds. Using NIR reflectance spectroscopy, blood stains were distinguished from other substances with 100% sensitivity and 100% specificity. In addition, Partial Least Squares Regression analysis was applied to estimate the age of blood stains on colored backgrounds. The age of blood stains up to 1 month old was estimated successfully with a root mean squared error of prediction of 8.9%. These findings are an important step toward the practical implementation of blood stain identification and age estimation in forensic casework, where a large variety of backgrounds can be encountered.     

Identification in blood stains through DNA typing with C4 and HLA-DR probes

A restriction fragment length polymorphism analysis using double digestion of DNA preparations with XbaI and BglII restriction enzymes and hybridization with C4 and HLA-DR probes is described. The typing conditions selected reveal extensive individual variation in both C4 and DR gene regions. In our panel of 46 unrelated individuals, 37 different phenotypic patterns were recognized when both probes were used, and preliminary discriminative power values of 0.865 and 0.914 were calculated for C4 and DR beta, respectively. The probability of a chance match using both systems is probably about 1.5.10(-2). The potential of this method for individual identification of blood stains was demonstrated on DNA prepared from 6-month-old dried blood stains from seven panel individuals. The seven individuals were all identified when comparing stain DNA patterns with panel control patterns. No RFLP pattern changes were observed following storage of blood stains. Based on these experiments with C4 and DR beta DNA typing under laboratory conditions, it is concluded that DNA typing with such probes may become a powerful tool in future stain identification analyses.     

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.     

Fluorescent TiO2 powders prepared using a new perylene diimide dye: applications in latent fingermark detection

A new, highly fluorescent dye was synthesised using oleylamine combined with a perylene dianhydride compound. The new dye was characterised by 1H NMR, UV-vis spectroscopy and fluorescence spectroscopy as well as quantum yield. The dye was absorbed onto titanium dioxide nanoparticles for use as a fingerprint detection powder. The new fluorescent powder was applied to latent fingermarks deposited onto different non-porous surfaces and compared with commercial fluorescent powders. The powder exhibits strong fluorescence at 650-700 nm under excitation at 505 nm. On glass surfaces, the new powder gave images showing tertiary-level detail of the fingermark ridges with almost no background development. Compared with current magnetic fluorescent powders, the new powder was slightly weaker in fluorescence intensity but produced significantly less background development, resulting in good contrast between the fingermark and the substrate.     

Latent evidence detection using a combination of near infrared and high dynamic range photography: an example using bloodstains

In this paper, we use bloodstains to illustrate an approach for identifying latent evidence on dark cloth using near infrared (NIR) photography combined with high dynamic range (HDR) photography techniques. NIR photography alone has been used to capture latent evidence that cannot be seen in normal ambient light. HDR techniques combine multiple bracketed photographs of the same image to increase the dynamic range of the photograph which can provide greater contrast. Using NIR photography alone, we were able to detect a bloodstain up to a 1/16 dilution, an improvement over previous studies. Combining NIR photography with the HDR process resulted in a noticeable increase in visibility up to 1/16 dilution when compared to NIR photographs alone. At 1/32 dilution, we were able to detect bloodstains that were not visible using NIR alone. NIR is a useful tool for imaging latent evidence, and combining NIR with HDR consistently provides better results over NIR alone.     

Visualization of dyes on thin-layer chromatography plates by means of the argon ion laser

A simple method for the detection of nanogram amounts of dyes by means of their fluorescence in the visible region of the spectrum is described. To stimulate the fluorescence, an argon ion laser with principal lines of 514.5 nm and 488.0 nm is used. The sensitivity of the technique allows the visualization of an amount of dye smaller than could be detected with either visible absorbance or ultraviolet stimulated fluorescence.     

A further study to investigate the detection and enhancement of latent fingerprints using visible absorption and luminescence chemical imaging

This study investigated the application of chemical imaging to the detection of latent fingerprints using the Condor macroscopic chemical imaging system (ChemImage Corp., Pittsburgh, USA). Methods were developed and optimised for the visualisation of untreated latent fingerprints and fingerprints processed with DFO, ninhydrin, cyanoacrylate, and cyanoacrylate plus rhodamine 6G stain. The results obtained with chemical imaging were compared to the detection achieved using conventional imaging techniques. The Condor significantly improved the detection of many prints, especially those that might be considered poor quality or borderline prints. Prints on newspaper treated with ninhydrin and DFO, and prints on white and yellow paper treated with ninhydrin, benefited the most from chemical imaging detection. In many cases, fingerprints undetectable using conventional imaging techniques could be visualised with chemical imaging. Ridge detail from untreated prints on yellow paper was also detected using the Condor. When prints of high quality were examined, both detection techniques produced quality results. The results of this project demonstrate that chemical imaging offers advantages over conventional visualisation techniques when examining latent fingerprints, especially those that would be considered difficult, such as weak prints or prints on surfaces that produce highly luminescent backgrounds. Standard testing procedures for the detection and enhancement of fingerprints by chemical imaging are presented and discussed.     

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.     

Identification of human urinary stains by the quotient uric acid/urea nitrogen

Uric acid (UA) and urea nitrogen (UN) were determined in urinary stains and the UA/UN x 20 values were calculated. The values in human urinary stains were 1.11-4.21, while those in other mammals except some of chimpanzees, were under 0.7, and those in fecal stains of birds were over 80. Most of the stains of other human body fluids or plant juices tested contained neither UA nor UN, and some contained one, but never the other. Ascorbic acid (AS) of up to 100 mg/dl in urine did not interfere with UA determination when dried human urinary stains were analyzed. It was also found that the contents of UA were very low at the peripheral parts of urinary stains. The present results indicate that the quotient UA/UN is useful for identification of human urinary stains in forensic practice provided that the peripheral part of the stain is not used.     

Differex~（TM）与Chelex-100法在混合斑检验中的应用比较

目的探讨DifferexTM系统法在法医学性侵犯案件混合斑检验中的应用价值,并与传统Chelex-100法进行比较。方法对日常性侵犯案件中的生物检材分别通过DifferexTM系统法与Chelex-100法检验来比较两种方法的优劣。结果采用Chelex-100法提取时,在所检验样本中仅有床单全部获得单一分型,而采用DifferexTM系统法则只有一条内裤的分型为混合型,其它均为单一分型。应用DifferexTM系统法还可以同时提取女性上皮细胞DNA,成功率为100%。采用Chelex-100法时出现双尖峰的频率也明显超过DifferexTM系统法。结论 DifferexTM系统法在法医学性侵犯案件混合斑检验中比Chelex-100法具有更明显的优势,是一种非常有应用前景的方法。     

Forensic Discrimination of Dyed Hair Color: I. UV‐Visible Microspectrophotometry*†

Abstract: Current protocols for examining hair do not attempt to differentiate hair color using instrumental analysis. In this study, hair samples treated with 55 different red hair dyes were analyzed using UV‐visible microspectrophotometry between 200 and 700 nm. Using air as a background reference gave the best results, although mounting media such as glycerin could also be used. The contribution of the hair substrate is predominantly observed in the range of 300–400 nm while the dye peak is evident in the range of 425–550 nm. It was found that the presence of hair dye reduces the overall intrasample variability of the hair color. In addition, visual inspection and spectral interpretation showed that dyed hair exhibits distinct and discernable shades. The color of all samples was stable during storage and while all hair dyes fade with washing, significant fading of the color was only evident after daily washing for 3 weeks.     

The application of visible wavelength reflectance hyperspectral imaging for the detection and identification of blood stains

Current methods of detection and identification of blood stains rely largely on visual examination followed by presumptive tests such as Kastle–Meyer, Leuco-malachite green or luminol. Although these tests are useful, they can produce false positives and can also have a negative impact on subsequent DNA tests. A novel application of visible wavelength reflectance hyperspectral imaging has been used for the detection and positive identification of blood stains in a non contact and non destructive manner on a range of coloured substrates. The identification of blood staining was based on the unique visible absorption spectrum of haemoglobin between 400 and 500 nm. Images illustrating successful discrimination of blood stains from nine red substances are included. It has also been possible to distinguish between blood and approximately 40 other reddish stains. The technique was also successfully used to detect latent blood stains deposited on white filter paper at dilutions of up to 1 in 512 folds and on red tissue at dilutions of up to 1 in 32 folds. Finally, in a blind trial, the method successfully detected and identified a total of 9 blood stains on a red T-shirt.     

新型荧光502粉末与502胶＋荧光染色显现潜在手印的比较

目的对新型荧光502粉末显现非渗透性光滑客体表面潜手印的一步荧光502显现新技术和传统502胶水熏显＋荧光染料染色显现手印技术进行比较研究。方法比较实验研究,对荧光502粉末和502胶水使用同一自动熏显柜先后进行熏显,荧光502粉末熏显结束后直接在蓝光灯下激发进行观察和拍照;502胶水在熏显完成和聚合物完全固化后使用罗丹明6G和BBD溶液进行荧光染色,再进行光致荧光照相。结果一步熏显和二次染色的显现效果基本相同,但两者的显现效果也因检材性质和遗留时间的不同而存在一些差别。结论新型荧光502粉末可以作为一种显现非渗透性光滑客体表面上潜手印的常用方法,尽管新方法对某些客体的显现效果并不好,但对大多数客体来说,其显现效果并不弱于二次染色的方法,并且使用新型荧光502粉末进行一步熏显不会引入有机溶剂,不会引起502聚合物的溶解或造成破坏,确保了纹线及其细节特征不被损坏,也避免了染色后因漂洗不当使手印纹线被冲掉的可能,还不会破坏脱落细胞等生物检材。     

Bloodstains on Leather: Examination of False Negatives in Presumptive Test and Human Hemoglobin Test

Presumptive tests for blood are very simple and sensitive tests used in the search for evidence. They also provide initial information on the nature of stains. A second test can confirm their nature. However, these tests can present false–negative results for different reasons. Some of those reasons have been studied, while others, those caused by the substrate material that contains the stain, are less well known. This work studies the effect of one component of a leather substrate—quebracho extract—on presumptive and human hemoglobin blood tests. Assays were performed using samples of blood dilutions contaminated with quebracho extract and others formed on a substrate containing the contaminant. Results show an undoubted interference that causes false negatives and even visible to the naked eye stains and also indicate that some tests (phenolphthalein) are more affected than others. Examiners should be taken into account when working on this kind of substrates.     

Evaluation of four deoxyribonucleic acid (DNA) extraction protocols for DNA yield and variation in restriction fragment length polymorphism (RFLP) sizes under varying gel conditions.

This study originated from discussions and recommendations of the Technical Working Group on DNA Analysis Methods (TWGDAM). Four bloodstain deoxyribonucleic acid (DNA) extraction protocols and five semen stain DNA extraction protocols were evaluated. Nine laboratories participated in the extraction of DNA from 20 bloodstains and 20 semen stains using each protocol. All blood and semen stains originated from a single donor and were prepared under uniform conditions to permit the direct comparison of DNA yields and restriction fragment lengths. The extracted DNA from approximately 600 bloodstains and 700 semen stains was quantified by yield gel analysis and a slot blot hybridization technique. The extracted DNA was digested and restriction fragment length polymorphism (RFLP) patterns were generated using three single-locus probes. The RFLP sizing data produced from the blood and semen stains were evaluated with respect to (1) DNA extraction method, (2) gel length, (3) agarose type, (4) presence or absence of ethidium bromide in the gel, and (5) fragment sizes obtained from DNA isolated directly from the donor's liquid blood. This study demonstrates conclusively that high-molecular-weight DNA can be isolated using either organic or nonorganic DNA extraction protocols and that the resulting RFLP sizes are highly reproducible regardless of gel length, agarose type, or presence/absence of ethidium bromide.     

pYNZ22位点扩增片段长度多态性法医应用的研究

对120名中国人进行pYNZ22位点扩增、银染检测，已找到11个等位基因，频率分布于0.4～30.4％，片段长170bp～870bp，相邻等位基因长度相差70bp，杂合度为73％，DP值为0．938。分析了五个家系均符合孟德尔定律。同一个体的血液、血斑、精液、混合斑中分离出的精子、毛囊、唾液及其它有核细胞组织的DNA多次进行pYNZ22位点扩增，得到一致的结果。灵敏度达到0．5ng核DNA。     

Evaluation of iodine-benzoflavone and ruthenium tetroxide spray reagents for the detection of latent fingermarks at the crime scene

The performance of two spray reagents, iodine-benzoflavone and ruthenium tetroxide (RTX), was evaluated and compared with the conventional technique currently used at the crime scene, that is, powdering. Neither the spray techniques nor powdering were shown to be suitable for all surfaces and ages of marks tested. On some surfaces such as glass and treated wood, powdering was still the superior technique, whereas the spray techniques produced better development on wallpaper, vinyl, and brick. Sequencing work showed that RTX was incompatible with powdering and cyanoacrylate (with a rhodamine 6G stain). Iodine-benzoflavone can be used successfully either before or after powdering in a sequence; however, it was incompatible with cyanoacrylate. Two non-CFC formulations of iodine-benzoflavone using HFC4310mee and HFE7100 solvents were tested and shown not to be as effective as the original Arklone (CFC-113) formulation; however, the HFC4310mee solvent is recommended as the most suitable replacement solvent. Due to the expense of the commercial RTX spray, attempts at formulating a more cost-effective version were also carried out. A formulation was developed that gave comparable development to the commercial version but at a much cheaper cost, and with a shelf life of up to two months. Recommendations are presented for which techniques are suitable for different surfaces and ages of marks. Powdering was shown to be the best technique on all ages of marks tested on treated wood, glass, and also on marks aged three days and older on paint. Iodine-benzoflavone was the best technique on wallpaper, vinyl, brick, and raw wood. RTX was the best detection technique for fresh marks and marks aged up to one day on wallpaper and paint.     

A BPA Approach to the Shroud of Turin

An investigation into the arm and body position required to obtain the blood pattern visible in the image of the Shroud of Turin was performed using a living volunteer. The two short rivulets on the back of the left hand of the Shroud are only consistent with a standing subject with arms at a ca 45° angle. This angle is different from that necessary for the forearm stains, which require nearly vertical arms for a standing subject. The BPA of blood visible on the frontal side of the chest (the lance wound) shows that the Shroud represents the bleeding in a realistic manner for a standing position while the stains at the back—of a supposed postmortem bleeding from the same wound for a supine corpse—are totally unrealistic. Simulation of bleeding from the nail wounds contacting wood surfaces yielded unclear results.     

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.