Workshops

Question 1

Give reasons why the inclusion of trace evidence in a criminal investigation may be beneficial.

Answer 1

• Trace evidence doesn’t lie, have an agenda, forget things, succumb to emotion or misinterpret scenarios.
• Traces can provide evidence of association or contact, between people, places and/or objects.
• They can assist in crime reconstruction, i.e. activity level propositions.
• They provide robust and complementary evidence in casework.
• We can no longer rely exclusively on DNA evidence as the sensitivity of its analysis means contamination is a problem.
• It can provide valuable evidence where DNA and fingermarks presence would be unhelpful. (For example a domestic assault case in the victims home)
• Where no matches are revealed on either the DNA or fingerprint databases, traces can provide investigative leads.

Question 2

What do we mean by the term chemometrics

Answer 2

• Computationally intensive, multivariate statistical analysis, applied to chemical systems or processes.
• Reduction of data dimensionality in complex datasets.
• Pattern recognition techniques that reveal sample groupings or underlying trends.
• An aid to objective and reproducible data interpretation or parameter optimisation.
• Techniques that reveal important variables within datasets and those that demonstrate covariance.
• An aid to data visualisation.

Question 3

Explain how Raman microspectroscopy is advantageous for the analysis of trace evidence specifically.

Answer 3

• It is a non-destructive technique so the sample’s integrity is maintained.
• It is a rapid technique.
• Microspectroscopy has a high spatial resolution so it is good for trace samples.
• Organic and inorganic components within a sample can be profiled simultaneously.
• It is a sensitive technique and is therefore able to detect slight changes in analyte concentration.
• It is superior to IR for the analysis of inorganic components, i.e. metal-ligand bonds.
• It aids identification of a sample through functional group assignments and provides additional, complementary information.
• Minimal sample preparation means sample integrity is maintained; sample alteration or contamination is unlikely.
• Polymorphs may be determined.
• Mapping function allows for sample heterogeneity to be ascertained.
• Microscope imaging allows precise focussing of laser on specific sites of interest within a sample.

Question 4

Name factors you would need to consider when interpreting the evidential value of fibres recovered from a deceased victim’s garment.

Answer 4

• Are they relevant in the context of the case?
• Do they “match” or share similar characteristics with anything relating to the suspect?
• Background levels – how common is the fibre?
• Must consider 2⁰, 3⁰ and even 4⁰ transfer.
• Non-contact transfer?
• The fibre’s material and shedding likelihood.
• The material of the garment.
• How many fibres were recovered.
• The location and surface area of fibre recovery.
• Any evidence of emergency services intervention?
• Any contamination from the scene?
• Have elimination samples been taken from emergency personnel and excluded as a source?
• Attempts at cleaning/laundering
• Environmental conditions at an indoor scene.
• Weather conditions at an outdoor scene.
• Wild animal interference? Or even household pet
• Time elapsed since crime to recovery and recovery to analysis.

Question 5

Name differences between raman and infrared

Raman

Answer 5

• Light scattering
• Requires change in polarisability (C=C, C-C, C-H)
• Little or no sample preparation necessary
• Measure through transparent packaging
• Aqueous samples

Question 6

Name differences between raman and infrared

IR

Answer 6

• IR absorption
• Requires change in dipole moment (O-H, N-H, C=O)
• Sample preparation usually necessary
• Short optical path length required
• Non-aqueous samples

Question 7

Which part of the analytical workflow for both fibre and paint evidence, would provide a result with the highest probative value?

Answer 7

• Physical fit

Question 8

In fibre analysis, name physical properties that we use microscopy to examine.

Answer 8

• Surface treatments
• Dye penetration
• Cross section
• Chemical composition
• Colorant/dye
• Texture
• Morphology
• Diameter
• Dichorism/pleochroism
• Isotopy/anisotropy
• Refractive index/birefringence

Question 9

Give pieces of information that are available for a particular colourant (pigment or dye) in the Colour Index International list.

Answer 9

• It’s application
• It’s hue
• It’s properties
• Chemical composition
• Generic name
• Colurant properties
• CI number
• Companies that manufacture it
• Trade names

Question 10

Explain why Principal Component Analysis (PCA) is a better unsupervised exploratory data analysis technique than Cluster Analysis (CA).

Answer 10

• CA groups samples into clusters based on their calculated distance (similarity), but the resultant dendrogram does not tell you why the samples are similar/different.
• PCA provides a loadings plot as part of the visual overview; this is a map of the variables, which in turn is related to the chemistry of the samples.
• The loadings plot can be used in conjunction with the scores plot - which is a map of the samples - to work out how the samples are similar/different based upon their chemical compositions.

Question 11

Equation for F

Answer 11

F = -kx

Question 12

Equation for stress

Answer 12

force / cross sectional area (A)

Question 13

Equation for strain

Answer 13

Extension/length

Question 14

Young modulus equation

Answer 14

• Plot stress on x axis
• Plot strain on y axis
• Gradient = young modulus

Question 15

What is the most common form of paint evidence?
When considering this evidence type with a comparison microscope, what particular features should you note?

Answer 15

• Automotive paint
• Colour of layers, thickness of layers, pigment distribution, textural chanes, layer sequence, pigment size and distribution, defects eg weathering, dirt and solvent traps

Question 16

What is the definition of the term “resolution”? What are the parameters that
limit the resolution of a light microscope?

Answer 16

• The limit of resolution in light microscopy is given by rayleighs criterion, half of the airy disk diameter (d) the airy disk diameter os given by d = 1.22 x wavelength x (focal length / lens diameter) acknoweldeg that wavelength, focal length and lens diameter matter for resolution.
• Resolution is the ability to determine between two points of specimen.

Question 17

In a light microscope explain what is the purpose of the… (i) Lamp at the base of the stand. (ii) Eyepiece lens.

Answer 17

1. Lamp in the base of the stand - Supplies light to the specimen. Without it the overall illumination and contrast would not be sufficient for imaging.
2. Eyepiece lens - Further magnifies the image from the objective and puts it in a form and position suitable for viewing.

Question 18

What is the role played by light in a light microscope?

Answer 18

In a reflected light microscope a light source (usually visible) is directed through a tube, reflects off the surface of the sample, and is then sent through a series of lenses to magnify the sample. The image is relayed to the eyepiece which puts it into a form suitable for viewing for the operator looking through the eyepiece.

Question 19

What are the different types of signal detected in SEM and typically what depth does each signal originate from, in relation to the surface of the sample? In your answer you should consider how these signals arise, following the interaction of the incoming probe beam with the sample.

Background

Answer 19

• SEM images are generally formed by detecting either the secondary SE (low-energy) electrons emitted from the sample, or the backscattered (high-energy) electrons. While secondary electrons can be formed anywhere within the volume of interaction, only those generated within a few nanometers of the surface actually escape to be detected. Most of these are generated before the beam spreads out, and thus provide information about a very small volume of material at the e-probe’s point of impact on the surface.
• Different types of signal originate from different depths.

Question 20

What is the role played by the beam of electrons in an Electron Microscope?

Answer 20

Electron microscopes use a beam of electrons rather than visible light to visualise the object. The beam interacts with the sample and produces several types of electron signals. Some of the electrons that are scattered are collected by a detector to produce an image, with magnifications of the order of 100,000x.

Question 21

What is the principal difference between SEM and TEM?

Answer 21

In a Scanning electron microscope (SEM) the focussed electron beam is scanned across the specimen and is reflected from the surface, while in a Transmission Electron Microscope (TEM) the beam passes through the specimen.

Question 22

What are the different types of signal detected in SEM and typically what depth does each signal originate from, in relation to the surface of the sample? In your answer you should consider how these signals arise, following the interaction of the incoming probe beam with the sample.

BSE

Answer 22

• The backscattered electrons able to escape from the sample are formed only in the first part of the electron trajectory, before the beam has lost too much energy, while they still stand a good probability of getting back to the surface, but still at greater depths than the secondary electrons.
• Hence the detected backscattered electrons (BSE) originate from a larger volume of the sample than do the secondary electrons, and thus form an image of lower resolution.
• BSEs that re-emerge from the sample surface can also generate secondary electrons, but in most cases these are few enough, and sufficiently diffuse, that they do not mask the high-resolution detail in the main secondary electron signal.

Question 23

What are the different types of signal detected in SEM and typically what depth does each signal originate from, in relation to the surface of the sample? In your answer you should consider how these signals arise, following the interaction of the incoming probe beam with the sample.

X-rays

Answer 23

• Production of X-rays can take place at any depth where the electron beam has enough energy to ionise the atoms.
• The probability of X-ray production will vary with depth, in a way that will vary from element to element, but it is clear that the volume from which the X-rays originate is substantially the whole interaction volume of the electron beam.
• While the X-rays can be absorbed as they pass through the material of the sample, they have, in general, a much longer path than electrons, and so have a good probability of leaving the sample and reaching the detector.
• Thus the spatial resolution of the X-ray signal is variable, and is frequently of the order of a micron or more.

Question 24

Introduction to pollen

Answer 24

• Pollen is the powder containing the male gametophytes of seed-producing plants - dispersed in order to reproduce by wind, water or animals.
• A single anther can produce 50-250,000 pollen grains depending on the plant species

Question 25

Pollen size

Answer 25

Generally less than 50μm in diameter (range 2-100μm), often with air bladders which allow them to keep aloft in the wind & travel up to 2km from the parent plant

Question 26

Pollen structure

Answer 26

- Have a fairly hard coat to protect them as they are transported and are relatively resistant to destruction - Surface texture can be incredibly diverse and identifiable to a particular plant

Question 27

Forensic Palynology

Answer 27

- Pollen’s small size, vast amount, resistance to destruction, ability to be dispersed easily & identifiable features make it excellent as trace evidence.

Question 28

Benefits of pollen

Answer 28

Further pollen persists for a long time (even after washing) due to excellent adhesion properties, making it likely to be found everywhere: Clothes, Hair, Drugs, Soil, Vehicles and even in stomach contents

Question 29

Drawback of pollen as trace evidence

Answer 29

Palynologists are skilled in identification and take advantage of these features, surveying plants when needed. This gives rise to 2 challenges: - Lack of skilled people in handling & identification - Limited national or international databases of pollen.

Question 30

Pollen analytical workflow

Answer 30

- Sample collection is key: Needs to be performed quickly but extremely carefully to avoid contamination. - Sample collection combined with site survey - Lack of skilled people raises issues of quality of samples- contamination risk is very high - Pollen chemically processed and prepared for identification methodology. - SEM considered the standard for analysis but this is time consuming when performed at scale. - Semi-automated TLM & fluorescence emerging as potential rapid, accurate alternative. - Molecular Barcoding with high throughput sequencing also emerging as future standard for analysis & identification.

Question 31

Pollen case study

Answer 31

- Samantha Hall - Murdered, body found in 1996 in remote bush-land near costal dunes in Queensland, Australia. - Estranged husband was primary suspect, but had recent washed all of his clothing thoroughly. - Despite washing, a few pollen grains from two different species of wattle bushes were found on his cleaned clothing. - One species was quite common to the region, but the other wasn’t native and had only been imported to help stabilise the costal dunes. - Despite rigorous denial by the suspect - pollen trace evidence was sufficient to help convict him of murder.

Question 32

Which part of the hair is largely responsible for its mechanical properties and colour?

Answer 32

- Cortext which is the main bulk of hair. - Largely responsible for the mechanical properties of hair. - Contains most of the pigment granules giving hair a colour.

Question 33

Describe pigment granule features that you could investigate for similarities with a compound light microscope.

Answer 33

* Density * Size * Shape * Aggregates * Distribution * Texture

Question 34

Based on the microscopical analysis of hair alone, what are the strengths and of hair as a form of trace evidence?

Answer 34

- Variation between individuals provides the discriminating potential for forensic hair comparisons. - It’s found on all humans and other mammals. - It’s easily overlooked by criminals involved in nefarious activities. - It’s highly stable, resisting both physical and chemical degradation. - It’s constantly being produced & shed in their immediate environments and readily transferred. - It’s highly stable, resisting both physical and chemical degradation

Question 35

Based on the microscopical analysis of hair alone, what are the weaknesses of hair as a form of trace evidence?

Answer 35

- hair cannot be individualised on the basis of these physical characteristics alone - using microscopy alone can’t determine if an unknown originates from a specific individual. - In almost all studies found that transfer and persistence of hairs are complex issues with far too many variables to be able to predict the absolute number of hairs expected to be transferred or remain on an object in a real‐world scenario.

Question 36

Give 3 reasons why glitter and shimmer, although both multilayered particles intended to add sparkle to cosmetic products, are very different.

Answer 36

* Glitter is synthetic; Shimmer is largely natural. * Glitter is plastic based (PET, PBT, PMMA), aluminium based, or a combination of the two; Shimmer is either mica based (muscovite or synthetic fluorphlogopite) or borosilicate glass based, and coated with various metal oxides. * Glitter is cut from rolled sheets into identical and specific shapes; Shimmer occurs as randomly sized and shaped flakes .

Question 37

You are presented with a small, transparent fragment of trace evidence, how would you go about confirming that it is glass through physical examination?

Answer 37

Quartz & Minerals - Birefringent Plastic - Compresses under pressure

Question 38

you identify that the fragment is quartz - describe in detail the optical methodology you would use to compare it to a known sample to help identify it.

Answer 38

Birefrigence and retardation measurements