Touch Samples Research Articles

A generic label-free microfluidic microobject sorter using a magnetic elastic diverter.

Cell sorters play important roles in biological and medical applications, such as cellular behavior study and disease diagnosis and therapy. This work presents a label-free microfluidic sorter that has a downstream-pointing magnetic elastic diverter. Different with most existing magnetic sorters, the proposed device does not require the target microobjects to be intrinsically magnetic or coated with magnetic particles, giving users more flexibility in sorting criteria. The diverter is wirelessly deformed by an applied magnetic field, and its deformation induces a fluid vortex that sorts incoming microobjects, e.g., cells, to the collection outlet. The diverter does not touch samples in this process, reducing the sample contamination and damage risks. This sorter uses a magnetic field generated by off-chip electromagnetic coils that are centimeters away from the device. With simple structure and no on-chip circuits or coils, this device can be integrated with other lab-on-a-chip instruments in a sealed chip, ameliorating the safety concerns in handling hazardous samples. The parallel and independent control of two such diverters on a single chip were demonstrated, showing the potential of doubling the overall throughput or forming a two-stage cascaded sorter. The sorter was modeled based on the Euler-Bernoulli beam theory and its reliability was demonstrated by achieving a raw success rate of 96.68% in sorting 1506 registered microbeads. With a simple structure, the sorter is easy and cheap to fabricate. The advantages of the proposed sorter make it a promising multi-purpose sorting tool in both academic and industrial applications.

Flow cytometry analysis of epithelial cell populations from touch samples using the BD Influx flow cytometry platform

'Touch' or trace cell mixtures submitted as evidence are a significant problem for forensic laboratories as they can render resulting genetic profiles difficult or even impossible to interpret. Optical signatures that distinguish epidermal cell populations from different contributors could facilitate the physical separation of mixture components prior to genetic analysis, and potentially the downstream production of single source profiles and/or simplified mixtures. For this dataset, optical properties including forwards scatter (FSC), side scatter (SSC), and fluorescence emissions in the Allophycocyanin (APC) channel were measured in epithelial cell populations from touch samples collected from several different contributors on multiple days to assess inter- and intra-contributor variability.

Screening Test for Shed Skin Cells by Measuring the Ratio of Human DNA to Staphylococcus epidermidis DNA.

A novel screening method for shed skin cells by detecting Staphylococcus epidermidis (S. epidermidis), which is a resident bacterium on skin, was developed. Staphylococcus epidermidis was detected using real-time PCR. Staphylococcus epidermidis was detected in all 20 human skin surface samples. Although not present in blood and urine samples, S. epidermidis was detected in 6 of 20 saliva samples, and 5 of 18 semen samples. The ratio of human DNA to S. epidermidisDNA was significantly smaller in human skin surface samples than in saliva and semen samples in which S. epidermidis was detected. Therefore, although skin cells could not be identified by detecting only S. epidermidis, they could be distinguished by measuring the S. epidermidis to human DNA ratio. This method could be applied to casework touch samples, which suggests that it is useful for screening whether skin cells and human DNA are present on potential evidentiary touch samples.

Forward-scatter and side-scatter dataset for epithelial cells from touch samples analyzed by flow cytometry.

‘Touch’ or trace biological samples submitted to caseworking labs as evidence often contain biological material from multiple individuals which can result in mixed DNA profiles. These mixture profiles are difficult to interpret and may cause analytical bottlenecks for forensic laboratories. The data in this brief reports the variation in the relative abundance of intact epithelial cells deposited by four different donors across nine days. Touch samples were generated each day by rubbing a polypropylene tube with both hands for five minutes. Forward-scatter area (FSC-A) and side-scatter area (SSC-A) data was acquired with the BD FACSCanto™ II Analyzer. The relative abundance of different sub-populations within the FSC-A and SSC-A plots was calculated against the total number of events analyzed in each sample. Mean and standard deviation values were calculated for each donor.

Optical characterization of epidermal cells and their relationship to DNA recovery from touch samples

The goal of this study was to investigate the relative contributions of different cellular and genetic components to biological samples created by touch or contact with a surface – one of the most challenging forms of forensic evidence. Touch samples were generated by having individuals hold an object for five minutes and analyzed for quantity of intact epidermal cells, extracellular DNA, and DNA from pelleted cell material after elution from the collection swab. Comparisons were made between samples where individuals had washed their hands immediately prior to handling and those where hand washing was not controlled. The vast majority (84-100%) of DNA detected in these touch samples was extracellular and was uncorrelated to the number of epidermal cells detected. Although little to no extracellular or cell pellet-associated DNA was detected when individuals washed their hands prior to substrate handling, we found that a significant number of epidermal cells (between ~5x10 3 and ~1x10 5) could still be recovered from these samples, suggesting that other types of biological information may be present even when no amplifiable nuclear DNA is present. These results help to elucidate the biological context for touch samples and characterize factors that may contribute to patterns of transfer and persistence of genetic material in forensic evidence.

Opticalcharacterization of epidermal cellsand their relationship to DNA recovery fromtouch samples.

The goal of this study was to investigate the relative contributions of different cellular and genetic components to biological samples created by touch or contact with a surface - one of the most challenging forms of forensic evidence. Touch samples were generated by having individuals hold an object for five minutes and analyzed for quantity of intact epidermal cells, extracellular DNA, and DNA from pelleted cell material after elution from the collection swab. Comparisons were made between samples where individuals had washed their hands immediately prior to handling and those where hand washing was not controlled. The vast majority (84-100%) of DNA detected in these touch samples was extracellular and was uncorrelated to the number of epidermal cells detected. Although little to no extracellular or cell pellet-associated DNA was detected when individuals washed their hands prior to substrate handling, we found that a significant number of epidermal cells (between ~5x10 (3) and ~1x10 (5)) could still be recovered from these samples, suggesting that other types of biological information may be present even when no amplifiable nuclear DNA is present. These results help to elucidate the biological context for touch samples and characterize factors that may contribute to patterns of transfer and persistence of genetic material in forensic evidence.

Assessment and mitigation of DNA loss utilizing centrifugal filtration devices.

Maximizing DNA recovery during its isolation can be vital in forensic casework, particularly when DNA yields are expected to be low, such as from touch samples. Many forensic laboratories utilize centrifugal filtration devices to purify and concentrate the DNA; however, DNA loss has been reported when using them. In this study, all centrifugal filtration devices tested caused substantial DNA loss, affecting low molecular weight DNA (PCR product) somewhat more than high molecular weight DNA. Strategies for mitigating DNA loss were then examined, including pre-treatment with glucose, glycogen, silicone (RainX(®)), bovine serum albumin, yeast RNA, or high molecular weight DNA. The length of pre-treatment and UV irradiation of pre-treatment reagents were also investigated. Pre-treatments with glucose and glycogen resulted in little or no improvement in DNA recovery, and most or all DNA was lost after silicone pre-treatment. Devices pre-treated with BSA produced irregular and uninterpretable quantitative PCR amplification curves for the DNA and internal PCR control. On the other hand, nucleic acid pre-treatments greatly improved recovery of all DNAs. Pre-treatment time and its UV irradiation did not influence DNA recovery. Overall, the results show that centrifugal filtration devices trap DNA, yet their proper pre-treatment can circumvent that loss, which is critical in the case of low copy forensic DNA samples.

Direct qPCR quantification of unprocessed forensic casework samples

The current short tandem repeat (STR) typing workflow for forensic casework samples involves sample collection, sample screening, DNA extraction, DNA qPCR quantification and STR amplification. Although very effective and powerful, this workflow still has room for improvements. For example, the screening assays in practice do not provide DNA related information and also do not work with touch DNA samples, which make up of the majority of the property crime samples. It is known that not all DNA samples have equal probative values. Considering the DNA backlog situation crime laboratories face today, an effective screening tool would be highly desirable. It would allow forensic scientists to prioritize the DNA samples so that the limited resources would be first spent on samples that would have better chances of producing informative STR profiles. qPCR assay does provide DNA quantity and gender information and would be an ideal screening tool. However, prior to quantification, sample extraction and purification are required. By the time a DNA sample is ready for qPCR quantification, time and resources have already been spent on samples that should have been given low priority or excluded from further processing if DNA quantity and gender information were known. To overcome this problem, a direct quantification technology is developed to allow qPCR quantification of casework samples without the need for DNA extraction and purification. The key to a direct qPCR assay is the PE-Swab, a novel sample collection device. A small sample punch can be generated from a PE-Swab and placed in a qPCR reaction for quantification. After optimizing the punch size and the quantification software baseline setting, accurate DNA quantification can be obtained from a sample without the need to carry out DNA extraction and purification. Proof of concept studies were done with low lever touch samples as well as blood samples. The PE-Swab also allows direct STR amplification of casework samples without the need for DNA extraction. Besides its potential as a screening tool, the direct qPCR assay can also be used to normalize the DNA input for a direct STR amplification reaction. The feasibility of the direct qPCR/direct STR amplification workflow was demonstrated with touch DNA samples and blood stain samples.

Development of a mRNA profiling multiplex for the inference of organ tissues

Forensic characterisation of organ tissue generally occurs through histological and immunological assays of limited sensitivity. Here, we explore an alternative approach and examine a total of 41 candidate mRNA markers for their ability to differentiate between brain, lung, liver, skeletal muscle, heart, kidney and skin. Various selection rounds are applied involving 85 organ tissues (36 excised autopsy specimens and 49 frozen tissue sections, with at least ten specimens for each organ type), 20 commercially available RNAs from different human tissues and at least two specimens of blood, saliva, semen, vaginal mucosa, menstrual secretion or touch samples. Finally, 14 markers are regarded tissue-specific and included in an endpoint RT-PCR multiplex together with one general muscle, one blood and one housekeeping marker. This 17-plex is successfully used to analyse a blind test set of 20 specimens including mixtures, and samples derived from stabbing of organ tissues. With the blind test set samples, it is shown that an earlier described interpretation strategy for RNA cell typing results [1] is also effective for tissue inference. As organ-typing is embedded in a procedure of combined DNA/RNA extraction and analysis, both donor and organ type information is derived from the same sample. Some autopsy specimens presented DNA profiles characteristic for degraded DNA. Nevertheless, the organ-typing multiplex could generate full RNA profiles, which is probably due to small sizes of the amplicons. This assay provides a novel tool for analysis of samples from violent crimes.

The Influence of Swabbing Solutions on DNA Recovery from Touch Samples,

There has been minimal research into how to best obtain DNA from touch samples. Many forensic laboratories simply moisten a swab with water and use it for collecting cells/DNA from evidentiary samples. However, this and other methods have not been objectively studied in order to maximize DNA yields. In this study, fingerprints were collected using swabs moistened with water or laboratory or commercially available detergents, including sodium dodecyl sulfate (SDS), Triton X-100, Tween 20, Formula 409(®) , and Simple Green(®) . Prints were swabbed, DNA isolated using an organic extraction, yields quantified, and relative yields compared. In all cases, the detergent-based swabbing solutions outperformed water, with SDS and Triton X-100 producing significant increases in yield. Short tandem repeat profiles were consistent with the individuals that placed them. Subsequent analysis of SDS concentrations for collecting touch DNA demonstrated an increase in DNA yield with increasing SDS concentration, with an optimal concentration of approximately 2%.

Specific and sensitive mRNA biomarkers for the identification of skin in ‘touch DNA’ evidence

In forensic casework analysis it is often necessary to attempt to obtain DNA profiles from microscopic amounts of biological material left behind by perpetrators of crime. The ability to obtain profiles from trace biological evidence is routinely demonstrated with so-called ‘touch DNA’ evidence, which is generally perceived to be the result of DNA obtained from shed skin cells transferred from donor to an object or person during physical contact. Although a genetic profile from trace biological evidence is routinely obtained, the tissue source of the profile is rarely known. This merely perpetuates the ‘mystery’ of the nature of ‘touch DNA’ evidence allowing the significance or meaningfulness of genetic profiles obtained from these samples to be challenged. Numerous reports state that the tissue source of origin of ‘touch DNA’ evidence cannot be determined due to the small amount of biological material present, while others conclude that the DNA profiles are obtained from shed skin cells (as opposed to, say, buccal epithelial cells present in saliva traces) without any scientific basis for this assertion. Proper identification of the biological material present might be crucial to the investigation and prosecution of a criminal offense and a misrepresentation of the nature of the evidence can have undue influence on the perception of the circumstance of the crime. Thus far, research has failed to provide forensic scientists with feasible, definitive methods to identify the tissue origin of ‘touch DNA’.In the present work, we sought to identify novel highly specific and sensitive messenger RNA (mRNA) biomarkers for the identification of skin. Gene candidates were identified using both literature searches and whole transcriptome deep sequencing (RNA-Seq). Utilizing this dual approach, we identified and evaluated over 100 gene candidates. Five mRNA markers were identified that demonstrated a high degree of specificity for skin. Using these markers, we have been able to successfully detect and identify skin using as little as 5–25pg of input total RNA from skin and, significantly, in swabs of human skin and various touched objects. One of the markers, LCE1C, is particularly highly sensitive and was detected in the majority of skin samples tested including touched objects. We have been successful in incorporating the five skin biomarkers into two multiplex systems. Although further work is needed to optimize the assay for routine casework, the initial studies demonstrate that a molecular-based characterization of the biological material recovered from touch samples is possible.

Identification of skin in touch/contact forensic samples by messenger RNA profiling

The true nature of touch DNA evidence has remained elusive, generally perceived to be the result of DNA obtained from shed skin cells yet never confirmed with scientific certitude. This is largely due to the belief that it is not possible to ascertain the tissue source of origin of the biological material in touch DNA evidence. Thus far, research has failed to provide crime laboratories with feasible methods to identify the tissue source of origin of touch DNA. The aim of the current work was to identify highly sensitive and specific biomarkers for the identification of skin. We have previously demonstrated the use of tissue specific messenger RNA (mRNA) profiling assays for body fluid identification. We therefore utilized mRNA profiling to identify potential biomarkers for the identification of skin. From an evaluation of over 100 potential genes, we identified five mRNA markers that demonstrated a high degree of specificity for skin. Using these markers, we have been able to successfully identify skin using as little as 5–25 pg of input RNA. The presence of skin has been successfully identified in swabs of human skin and in a variety of touch samples. One of the markers (LCE1C) is particularly highly sensitive and permits the detection of skin in a majority of known skin containing samples tested. Although further work is needed to produce an assay for routine casework, these initial studies demonstrate that a molecular-based characterization of the biological material recovered from touch samples is possible.

Froth touch samples viewed with Scanning Electron Microscopy

The stability of the thin films found in the froth phase of flotation plays an important role in the overall efficiency of the separation process. In turn, the physical behaviour of the particles attached to the films has a direct impact on the lamella stability. This effect is not fully understood. The film solids loading, or attached mass to area ratio, is an indicator of froth stability and has been measured by touch sampling of surface bubbles. Multiple samples can also be assayed as an indicator of maximum possible grade. Here the touch sample technique is used to show the particle packing arrangement on the film. This work presents images of touch samples from the top of the froth viewed under a Scanning Electron Microscope (SEM). Froth samples were obtained during batch flotation of glass beads with a size fraction of 90–150 μm. The packing arrangement of the particles attached to the surface lamellae were clearly resolved, making it possible to obtain a measure of film packing density using image analysis. These results can be used to validate and expand physical models that predict film stability.

Consumer product evaluation based on tactile sensory information

Abstract This paper presents the results of an experiment in which consumers were asked to examine visually and touch samples of fabric. The participants were either given or not given verbal information relevant to the product category, and provided answers to the dependent variables immediately or after a 4‐minute delay. The results showed that memory, confidence in memory, and confidence in product evaluation were positively influenced by the availability of verbal information and therefore support the proposition that relevant verbal information can improve the encoding, retention, and retrieval of consumer experiences that include sensory information and lead to preferences that are better defined. Copyright © 2010 John Wiley & Sons, Ltd.

Task planning for dextrous manipulation using blind grasping tele-assistance

An approach to operator interaction for a dextrous manipulator is described, using a tele-assistance method termed blind grasping. Given approximate initial coordinates for the object location, the blind grasping approach sequences a series of dextrous finger contact movements to establish the object geometry and candidate stable grasp locations. The approach is implemented within the framework of the preference planning tele-assistant task planning architecture. Geometric and semantic models of the world are used as triggers into pre-stored lists of action sequences. Actions are triggered when their pre-conditions fail, representing a state in the world that the action can correct. Failure of the action to change the world state hierarchically triggers sub-actions, until atomic actions instruct lower levels of control. For the first prototype AMADEVS dextrous underwater gripper, it is demonstrated that variously shaped objects centred between the open gripper fingers can generally be grasped using simple closure. However, off-centre displacements as small as 1 cm require five touch samples to acquire the geometry with sufficient accuracy to choose stable grasp contact points.

Experimental study of probe–surface interaction in near-field optical microscopy

The physical mechanism of shear-force interaction between a fiber tip and a sample in near-field optical microscopy has been studied using tunneling current measurements between the tip and a sample. Optical and non-optical (tuning fork) shear-force distance controls have been used. In both cases a tunneling current between the tip and the sample has been detected at the very beginning of shear-force approach curve when the amplitude of tip vibration was just slightly smaller than its initial value. These results indicate that fiber-tips generally mechanically touch samples under shear-force distance feed-back control.