Ifugao State University

COLLEGE OF CRIMINAL JUSTICE EDUCATION

Potia Campus, Alfonso Lista Ifugao

INSTRUCTIONAL MATERIALS in FORENSIC INSTRUMENTATION

I. Introduction of Forensic Science Instrumentation

Forensic
 The adjective forensic comes from the Latin word forensis, meaning “in open
court” or “public.” When you describe something as forensic you usually mean
that is has to do with finding evidence to solve a crime. It could also mean that it
has to do with the courts or legal system. Forum
 Relating to or denoting the application of scientific methods and techniques to the
investigation of crime.
 Scientific tests or techniques used in connection with the detection of crime.
 Forensic science, or forensics, is the application of science to establish how
historical events occurred and thereby provide impartial evidence that can be
used in a court of law.
Forensic science was used to solve important issues and criminal cases
long before the term was coined. Famous examples include Archimedes’
discovery that the amount of gold used in King Hiero II’s crown could be
calculated by immersing the crown in water and measuring the amount of water it
displaced, and the autopsy of the Roman Emperor Julius Caesar in 44 BCE that
confirmed his death by blood loss.
Forensic Instrumentation
 Forensic science instrumentation is a branch of pure chemistry having similarities
to physical chemistry.
 It is a branch in which new methods and instrumentation are applied on the
evidences to get the nature, information and compositions of the matter of
evidences.
- Range of Instrumentation
Forensic laboratory equipment ranges from instrumentation you would see
in a general laboratory, such as microscopes, fume hoods, chromatographs and
spectrometers, to equipment used for specific forensic analysis, like
cyanoacrylate fuming chambers for lifting of latent fingerprints. Alternative light
sources are used frequently both at the crime scene and in the laboratory, in
addition to cameras and video recording devices. Analysis of digital evidence,
like phones and computers, requires high-tech extraction software and analysis
tools.

What is a forensic investigation?

Forensics are the scientific methods used to solve a crime. Forensic
investigation is the gathering and analysis of all crime-related physical evidence
in order to come to a conclusion about a suspect. Investigators will look at blood,
fluid, or fingerprints, residue, hard drives, computers, or other technology to
 establish how a crime took place. This is a general definition, though, since there
are number of different types of forensics.

Forensic Laboratory Equipment

When equipping a forensic laboratory, the cost of purchasing and maintaining
instrumentation and equipment is always the first considerations. In a forensic lab, the
second tends to be analysis runtime. Due to increasing reliance on forensic analysis in
criminal investigations—and a lack of personnel—many forensic labs have faced
mounting workloads and backlogs. Employing instrumentation and equipment that is
automated, fast and accurate allows forensic professionals to move on to the next case
as quickly as possible.
Importance of Forensic Instrumentation
 Develop an understanding of those instrumental tools and their application to
solve important analytical problems.
 Familiarize with the fundamental principles of operation of modern analytical
instrumentation;
 Appropriate choices and efficient use of measurement tools;
 Knowledge of measurement principles is necessary for calibration,
standardization and validation of instrument method.

Uses of forensic science instrumentation

Instrumentation have several uses in the field of forensic science. Some of them
are:
 It helps to separate out the components of evidence.
 Separates out drugs from sample in less time (from body, or in pharmacy)
 Separation of ink.
 Poison detection.
 Used in water treatment
 Microscopic technologies are used in analysis of questioned documents.
 Separation of blood components.
 Analysis of metal
 Recognizing the bombs
 Identification of gunshot residue
 Analysis of paints.

Laws and Principles of Forensic Science

Forensic Science is the science which has developed its own Laws and
Principles. The Laws and Principles of all the natural sciences are the bases of forensic
Science.
Every object, natural or man-made, has an individuality which is not duplicated in
any other object.
1. Law of Individuality
Anything and everything involved in a crime, has an individuality. If
the same is established, it connects the rime and criminal.
 This principle at first sight appears to be contrary to common beliefs
and observations. The grains of sand or common salt, seeds of plants or
twins look exactly alike.

2. Principle of exchange
Contact exchange traces is principle of exchange. It was first
enunciated by the French scientist, Edmond Locard. Commonly known as
Edmond Locard’s maxim on Interchange.
According to the principle, when criminal or his instruments of crime
come in contact with the victim or the objects surrounding him, they leave
traces. Likewise, the criminal or his instruments pick up traces from the
same contact.

3. Law of Progressive change

Change is inevitable’’, this also applies to object. Different type of
objects may take different time spans.
The criminal undergoes progressive changes. If he is not
apprehended in time, he becomes unrecognizable. The scene of
occurrence undergoes rapid changes. The weather, the vegetable growth,
and the living beings make extensive changes in comparatively short
periods. Sample degrade with time, bodies decompose, tire tracks & bite
marks fade, the firearm barrel loosen, metal object rust, etc.

4. Principle of Comparison
Only the likes can be compared’’ is the principle of comparison. It
emphasize the necessity of providing like samples and specimens for
comparisons with the questioned items. A question hair can only be
compared to another hair sample, same tool marks, bite marks, tire marks,
etc.

5. Principle of Analysis
The analysis can no better than the sample analyzed. Improper sampling
and contamination render the best analysis useless. The principle
emphasizes the necessity of correct sampling and correct for effective
experts.

6. Law of Probability
All identification, definite or indefinite, are made consciously or
unconsciously, on the basis of probability.

7. Law of circumstantial facts

Evidences given by eye witness or victim may not always be accurate.
Sometimes victims may intentionally lie or sometimes because of poor
sense (such as low sight, unclear hearing), exaggeration & assumptions.
Important terminologies associated with instrument
1. Analytical Technique – It is vital scientific phenomenon which prove to be useful
for providing information regarding composition of a substance.
2. Analytical method - It is definite application of a method to analyses a sample
- It is also known as instrumental method

Classification of instrumental techniques

1. Microscopy
2. Spectrometry
3. NAA-Neutron Activation Analysis
4. X-ray & X-ray based techniques
5. Mass spectroscopy
6. Chromatography
7. Electrophoresis
8. Immunoassays

Classification of Method
Deals with method for determining the chemical composition of samples (evidence)
of matter as applies to forensic problems.
A. Classical Method
- In the early years of chemistry, most analyses were carried out by
separating the components of interest in a sample by precipitation,
extraction or distillation.
- The majority of classical analytical methods rely on chemical reactions
to perform an analysis.

a) Qualitative Analysis
- Yields information about the identity of atomic or molecular species or
the functional groups in the sample.
- the separated components were then treated with reagents that yield
products that could be recognized by their colors, their boiling and
melting points, their solubilities in a series of solvents, their odors, their
optical activities or their refractive indexes

1. Chemical Test- A chemical test is typically a fast reaction

performed in a test tube that gives a dramatic visual clue (a
color change, precipitate, or gas formation) as evidence for a
chemical reaction.
2. Flame Test – used to identify the presence of a relatively small
number of metal ions in a compound. Not all metal ions give
flame colours.

b) Quantitative Analysis
o Provides numerical information as to the relative amount of one or
more components of the sample.
o The amount of analyte is determined by gravimetric or by volumetric
measurements.
1. In gravimetric measurements, the mass of the analyte or some
compound produces from the analyte is determined.
 2. In volumetric, also called titrimetric procedures, the volume or
mass of a standard reagent required to react completely with
the analyte is measured

B. Instrumental Method
The instrumental methods of chemical analysis are divided into categories
according to the property of the analyte that is to be measured. Many of the
methods can be used for both qualitative and quantitative analysis. The major
types of instrumental methods are the spectral, electroanalytical, and
separatory.
a. Spectral methods
Spectral methods measure the electromagnetic radiation that is
absorbed, scattered, or emitted by the analyte. Because the types of
radiation that can be monitored are multitudinous and the manner in which
the radiation is measured can significantly vary from one method to
another, the spectral methods constitute the largest category of
instrumental methods.

b. Electroanalysis
The second major category of instrumental analysis is
electroanalysis. The electroanalytical methods use electrically conductive
probes, called electrodes, to make electrical contact with the analyte
solution

c. Separatory methods
The final major category of instrumental methods is
the separatory methods. Chromatography and mass spectrometry are two
such methods that are particularly important for chemical analysis.

Types of Forensic Analysis

While the structure of forensic labs can vary, especially between private, public
and government, a fully funded forensic laboratory often includes the following sections:
 Biology / DNA Analysis
 Drug Chemistry
 Forensic Toxicology
 Firearms / Tool marks
 Latent Prints
 Trace Evidence Analysis
 Footwear / Tire Tracks

Certain forensic labs may also include more niche analysis sections, such as:
 Questioned documents
 Microscopy
 Crime Scene and Crash Reconstruction
 Digital Evidence
 Instrumentation and Analytical Methodology in Forensic Science
Forensic analysis generally consists of three different components which may be
employed in any given case.
1. Identification refers to the qualitative determination of the identity of a substance,
based on measurements of its physical and chemical properties. Results of such
identifications alone do not usually shed much new light on a case; however,
there are cases in which the identification of a substance completes the forensic
investigation. A controlled substance possession case is a good example of the
latter. Not only is identification of the substance all that is required of the analyst,
but also it is an essential legal requirement if the charge is to be sustained.
2. Individualization refers to the demonstration of common origin between two
objects. At present, not many types of evidence can be completely individualized.
A true individualization is a demonstration that a sample or item is unique within
its own class or that questioned and known samples had a common origin.
3. Reconstruction refers to the establishment of the place, time, and sequence of
events that occurred during the perpetration of a criminal act. Complete forensic
analysis consists of: (1) recognition of the physical evidence; (2) collection of the
physical evidence, observing appropriate preservation and documentation
procedures; (3) submission of the evidence for analysis; (4) analysis of the
evidence, which may involve identification and/or individualization procedures;
(5) interpretation of the results of the examination in terms of the facts of the
case; and (6) an attempt to reconstruct the events and present the results and
conclusions to appropriate investigative and judicial bodies.

II. Instrumentation Method -

1. Microscopy
- A microscope is an optical instrument that uses a lense or combination
of lenses to magnify and resolve the fine details of an object. The
earliest methods for examining physical evidence in crime laboratories
relied almost solely on the microscope to study the structure and
composition of matter.
- The microscope is used by forensic scientists to locate, isolate,
identify, and compare samples. Because of its low magnification, wide
field of view, large working distance, and stereoscopic vision, the
stereomicroscope is used for preliminary evidence evaluations.
The history of the microscope spans centuries.
Roman philosophers mentioned “burning glasses” in their writings
but the first primitive microscope was not made until the late 1300’s. Two
lenses were placed at opposite ends of a tube

First Microscope
Grinding glass to use for spectacles and magnifying glasses
was common place during the 13th century. In the late 16th century
several Dutch lens makers designed devices that magnified objects. Dutch
spectacle makers Zaccharias Janssen and Hans Lipperhey are noted as
the first men to develop the concept of the compound microscope. But in
1609 Galileo Galilei perfected the first device known as a microscope.

Lens Improvement
Later in the 16th century, Anton van Leeuwenhoek began polishing
and grinding lenses when he discovered that certain shaped lenses
increased an image’s size.
Leeuwenhoek is considered the founder of the study of microscopy
and played a vital role in the development of cell theory.

Achromatic Lens
Using early microscopes was difficult. Light refracted when passing
through the lenses and altered what the image looked like.
When the achromatic lens was developed for use in eyeglasses by
Chester Moore Hall in 1729, the quality of microscopes improved.

Mechanical Improvements
During the 18th and 19th centuries, many changes occurred in both
the housing design and the quality of microscopes.
Microscopes became more stable and smaller. Lens improvements
solved many of the optical problems that were common in earlier versions.

August Kohler is credited with inventing a way to provide uniform

microscope illumination that allowed specimens to be photographed.

Ernst Leitz devised a way to allow for different magnifications

using one microscope by putting multiple lenses on a movable turret at the
end of the lens tube.
Looking for a way to allow more light-spectrum colors to be visible,
Ernst Abbe designed a microscope that in a few years would provide
Zeiss with the tools to develop the ultraviolet microscope.

Modern Technology Improving Microscopy

The invention of the microscope allowed scientists and scholars to
study the microscopic creatures in the world around them. Charles
Spencer demonstrated that light affected how images were seen. It took
over one hundred years to develop a microscope that worked without light.

Electron Microscopy - is a microscope that focuses beams of

energetic electrons to examine objects up to nano-scales.

Dr Sydney Smith, in 1934, was the first person to use microscopic hair
comparisons in a murder trial by visually matching hair from the crime
scene to the hair of the defendant. By the late 1970s, FBI laboratory
personnel commonly used microscopic hair analysis as part of their toolkit
in forensic investigations
 Virtual vs Real Images - Virtual image: refers to an image that is only
observable through a lenses or series of lenses.
Real image: refers to an image that is not observed through a lenses.
The magnifying glass - A single lenses generally held in the hand that can
magnify an image about five to ten times.
Types of Microscopes - The optical principles of the compound microscope are
incorporated into the basic design of different types of light microscopes.
A. The compound microscope
A compound microscope is a type of microscope which uses visible light
and a system of lenses to magnify images of small samples.
Compound microscopes are designed to view specimens that are
transparent -- they have been stained and affixed to a slide.
Objective Lenses:
Usually you will find 3 or 4 objective lenses on a microscope.
Our microscopes consist of 4x, 10x, 40x powers.
When coupled with a 10x (most common) OCULAR -eyepiece lens, total
magnification is
40x = (4x times 10x)
100x = (10x times 10x)
400x = (40x times 10x)

The mechanical system is composed of six parts:

Base - the support upon which the instrument rests.
Arm - a C-shaped upright structure, hinged to the base that supports the
microscope.
Stage - The horizontal plate upon which the specimen rests
Body Tube - which houses the objective and focus lenses
Course and Fine adjustments - to bring the lenses into alignment.

The optical system is made up of four parts:

Illuminator, artificial light source used to illuminate the specimen being examined.
Condenser, collects light rays from the illuminator and concentrates them on the
specimen.
Objective lense, is the lense positioned closest to the specimen.
Eyepiece, Ocular lense, is the lense closest to the eye.

B. The comparison microscope

 Modern firearms (BALLISTICS) examination began with the introduction of
the comparison microscope, with its ability to give the firearms examiner a
side-by-side magnified view of bullets.
 Essential to the use and application of the comparison microscope is the
closely matched nature of the objective lenses with minimal but identical lens
distortions

C. The stereoscopic microscope – The Dissecting Microscope

  The stereoscopic microscope has proven most useful for the examination of
details that characterize many types of physical evidence, not requiring high
magnification.
 The stereoscopic microscope provides magnifying powers that range from
10x to 125x
 The stereoscopic microscope has the distinct advantage of offering a three-
dimensional image of an object. It also provides a right-side-up image.
 The stereoscopic microscope has a wide field of view and offers great depth
of focus, making it an ideal instrument for locating trace evidence that may be
associated with debris, garments, weapons, or tools.
 Stereoscopes are able to view non-transparent objects at much lower
magnifications than compound microscopes. The lower magnification of a
stereoscope is not a shortcoming but a design decision. Stereoscopes are
used to view larger, three dimensional objects.

D. The polarizing microscope

 In 1808, French physicist Etienne Louis Malus discovered the refraction
and polarization of light.
 In 1828, William Nicol of Edinburgh invented a prism for polarization, as
an indispensable part of polarizing.
 In 1839, William Henry Fox Talbot invented polarizing microscope
 A compound or stereoscopic microscope can be modified to be outfitted
with a polarizer and analyzer so as to be capable of allowing the viewer to
detect polarized light.
 The effect of introducing a specimen that polarizes light will be to orient
the polarized light, allowing it to pass through the analyzer. The result
produces vivid colors and intensity contrast that make the specimen
readily distinguishable. Usually used in the field of geology.
 Produces 3D images
 Works on the principle of interference
 Polarizer, positioned in the light path somewhere before the
specimen, and:
Analyzer (a second polarizer), placed in the optical pathway
between the objective rear aperture and the observation tubes or camera
port. Image contrast arises from the interaction of plane-polarized light
with;
Birefringent (or doubly-refracting) specimen to produce two
individual wave components that are each polarized in mutually
perpendicular planes. The velocities of these components, which are
termed the ordinary and the extraordinary wavefronts, are different and
vary with the propagation direction through the specimen. After exiting the
specimen, the light components become out of phase, but are recombined
with constructive and destructive interference when they pass through the
analyzer.

E. The micro spectrophotometer

 First microspectrophotometer was built in the 1940s.
 By linking a microscope to a computerized spectrophotometer, a new
dimension has been added to the capability of the microscope, giving rise
to the micro spectrophotometer.
  Using the micro spectrophotometer, a forensic analyst can now view a
particle under a microscope while, at the same time, a beam of light is
directed at the particle in order to obtain its absorption spectrum.
 Designed to measure UV-Visible-NIR spectra of microscopic samples or
microscopic areas of larger objects. (Note: NIR= near infrared)
 To measure UV-visible-NIR range transmission, absorbance, reflectance,
emission and fluorescence spectra of sample, you need the CRAIC
software.
o This allows identification of
- Dyes
- Materials
- Concentration of chemicals

ELECTRON MICROSCOPE
In 1931, German scientist Ernst Ruska developed the first electron microscope.
The Electron Microscope (EM) is an impressively powerful microscope that exists
today, allowing researchers to view a specimen at nanometer size.
They utilize the same principles behind an optical microscope, but rather than photons
or particles of light, concentrate electrons, charged particles located on the outside of
atoms, onto an object.
Additional differences include preparation of specimens before being placed in the
vacuum chamber, the use of coiled electromagnets instead of glass lenses, the use of a
thermionic gun as an electron source and the image or electron micrograph is viewed
on a screen rather than an eyepiece.
Techniques, which vary based on type of specimen and analysis, include:

 Cryofixation - a technique for fixation or stabilization of biological

materials as the first step in specimen preparation for electron microscopy
and cryo-electron microscopy.
 Fixation - Fixation of tissues is the most crucial step in the preparation of tissue
for observation in the transmission electron microscope.
a. Osmium tetroxide and aldehydes
b. Sodium acetate/sodium veronal
 Dehydration – remove water from the biological specimens so that they
can be further processed using light microscopy and electron microscopy.
 Embedding – tissues to be analyzed in the electron microscope are
usually embedded in resins for ultramicrotomy.
 Epoxy resins
 Acrylic resins
 Sectioning - produce clear images of focal planes deep within a thick
sample.
 Staining - the most widely used stains in electron microscopy are
the heavy metals, uranium and lead.
  Freeze-fracture and Freeze-etch - a process whereby specimens,
typically biological or nanomaterial in nature, are frozen, fractured, and
replicated to generate a carbon/platinum “cast” intended for examination
by transmission electron microscopy
 Sputter Coating - is the process of applying an ultra-thin coating of
electrically-conducting metal – such as gold (Au), gold/palladium (Au/Pd),
platinum (Pt), silver (Ag), chromium (Cr) or iridium (Ir) onto a non-
conducting or poorly conducting specimen.
F. Transmission Electron Microscope (TEM)
Like the scanning electron microscope, the transmission electron
microscope (TEM) uses electrons in creating a magnified image, and samples
are scanned in a vacuum so they must be specially prepared. Unlike the SEM,
however, the TEM uses a slide preparation to obtain a 2-D view of specimens,
so it's more suited for viewing objects with some degree of transparency. A
TEM offers a high degree of both magnification and resolution, making it useful
in the physical and biological sciences, metallurgy, nanotechnology and
forensic analysis.
The TEM is a popular choice for nanotechnology as well as semiconductor
analysis and production.

G. The Scanning Electron Microscope (SEM) is a different and final approach to

microscopy that will be considered.
The image formed by the scanning electron microscope is produced by
targeting, with electromagnets, a beam of electrons onto the specimen and
studying the electron emission on a closed TV circuit. The primary electron
beam, emitted from a hot tungsten filament, causes the emission of electrons
from the element making up the outer layer of the specimen. The emitted
electrons are collected, and the integrated and amplified signal is displayed on
the TV circuit.

- The major attractions of the SEM image are its high magnification, high
resolution, and great depth of focus.
- In its usual mode the SEM has a magnification range of 10X to 100,000X.
- Its depth of focus is some 300X better than optical systems at similar
magnifications, and the resultant picture is almost stereoscopic in
appearance.
- Its great depth of field and magnification are invaluable in determining
structural relationships over a contextually broad area.
- Microscope examination of fibers, like the cotton fiber using SEM, cannot
provide quantitative characteristics of fibers, but is non-destructive and can be
used for a qualitative comparison.
Although SEMs are approximately 10 times less powerful than TEMs, they
produce high-resolution, sharp, black and white 3D images.

Electron Microscope Advantages

 The primary advantage is its powerful magnification.
 The potential runs the gamut of scientific fields including biology, gemology,
medical and forensic sciences, metallurgy and nanotechnologies.
  EMs also have many technological and industrial applications, such as
semiconductor inspection, computer chip manufacturing, quality control and can
even be used as part of a production line.

Electron Microscope Disadvantages

 The main disadvantages are cost, size, maintenance, researcher training and
image artifacts resulting from specimen preparation.
 This type of microscope is a large, cumbersome, expensive piece of equipment,
extremely sensitive to vibration and external magnetic fields.
 It needs to be kept in an area large enough to contain the microscope as well as
protect and avoid any unintended influence on the electrons.
 Upkeep involves maintaining stable voltage supplies, currents to electromagnetic
coils/lens and circulation of cool water so the samples are not damaged or
destroyed from heat given off during the process of energizing the electrons.
 Special training is required to learn the involved processes of specimen
preparation, to minimize and recognize preparation-related artifacts and to
operate the microscope itself.
 Despite these disadvantages, EMs are assets to high-end research laboratories;
this powerful piece of equipment has resulted in innumerable advances in
science and industry.

2. Spectroscopy

2.1 Spectroscopy - is the study of the absorption and emission of light and other
radiation by matter. It involves the splitting of light (or more precisely electromagnetic
radiation) into its constituent wavelengths (a spectrum), which is done in much the same
way as a prism splits light into a rainbow of colours. In fact, old style spectroscopy was
carried out using a prism and photographic plates.
 a. Absorption
- It is a technique in which the power of a beam of light measured before and
after interaction with a sample is compared.
Example: Infrared Spectroscopy (IR), MRI etc.
b. Emission
- It uses the range of electromagnetic spectra in which a substance radiates
(emits)
- The substance first must absorb
Some practical ways we use spectroscopy include:
 We can use the unique spectra to identify the chemical makeup, and
temperature and velocity of objects in space.
 For metabolite screening and analyzing, and improving the structure of
drugs.
 For measuring sampled chemicals or nanoparticles through their mass-to-
charge ratio using a mass spectrometer.

2.2 Spectrometry - is the measurement of the interactions between light and matter,
and the reactions and measurements of radiation intensity and wavelength. In other
words, spectrometry is a method of studying and measuring a specific spectrum, and
it’s widely used for the spectroscopic analysis of sample materials.

History of Spectrometry
- 1600s Isaac Newton discovered that focusing light through glass
split it into the different colours of the rainbow (known as the spectrum of visible
light).
- Generations of work by scientists, such as William Hyde Wollaston,
lead to the discovery of dark lines that were seemingly randomly placed along
 this spectrum. Eventually it was determined that these were the after-effects of
the absorption of chemicals in the earth’s atmosphere.
- 1814, Joseph von Fraunhofer invented the spectroscope and he is
sometimes referred to as the father of the spectroscopy.

As each atom corresponds to and can be represented by an individual

spectra, we can use the analysis of wavelengths in the light spectrum to identify
them, quantify physical properties, and analyze chemical chains and reactions
from within their framework.

A spectrometer consists of three main components – entrance slit, grating and detector.
a. Entrance Slit
Light from the source enters the entrance slit and the size of the slit determines
the amount of light that can be measured by the instrument. The slit size also affects
the optical resolution of the spectrometer, where the smaller the slit size, the better
the resolution.
b. Grating
A monochromatic uses a phenomenon of optical dispersion in a prism or
diffraction from diffraction gratings to select a particular wavelength of light. In
traditional spectrometers, prisms were used to disperse light.
c. Detector
The detector captures the light spectra and measures the intensity of light as a
function of wavelength. These data are then digitized and plotted onto a software as
a graph.

Types of Spectrometer
1. Nuclear Magnetic Resonance (NMR) Spectrometer
- The NMR Spectrometer observes and measures the interaction of nuclei spins when
the sample is placed in a strong, constant magnetic field. The NMR signal is produced
when the nuclei interacts with the magnetic field at a frequency that resonates with the
frequency of the nuclei.

2. Mass Spectrometer
- A Mass Spectrometer measures the mass-to-charge ratio of ions and identifies the
composition of elements present in a sample. This works by ionizing a sample, which
causes some of the molecules to become charged and separate according to their
mass-to-charge ratio.
These ions are then detected by a device that can detect charged particles.

3. Optical Spectrometer
- An Optical Spectrometer measures the properties of light, usually near the optical
region in the electromagnetic spectrum, i.e. ultraviolet, visible and infrared light.

4. Atomic Absorption Spectrometry (AAS)

Atomic absorption spectrometry (AAS) detects elements in either liquid or solid samples
through the application of characteristic wavelengths of electromagnetic radiation from a
light source. Individual elements will absorb wavelengths differently, and these
absorbance are measured against standards. In effect, AAS takes advantage of the
different radiation wavelengths that are absorbed by different atoms.

Differences between Spectrometry and Spectroscopy

 Spectroscopy is the science of studying the interaction between matter
and radiated energy. It’s the study of absorption characteristics of matter, or
absorption behavior of matter, when subjected to electromagnetic radiation.
Spectroscopy doesn’t generate any results, it’s simply the theoretical approach to
science.
On the other hand, spectrometry is the method used to acquire a
quantitative measurement of the spectrum. It’s the practical application where
results are generated, helping in the quantification of, for example, absorbance,
optical density or transmittance.
In short, spectroscopy is the theoretical science, and spectrometry is
the practical measurement in the balancing of matter in atomic and
molecular levels.

3. Chromatography
Is an important biophysical technique that enables the separation,
identification, and purification of the components of a mixture for qualitative and
quantitative analysis. Proteins can be purified based on characteristics such as
size and shape, total charge, hydrophobic groups present on the surface, and
binding capacity with the stationary phase. Four separation techniques based on
molecular characteristics and interaction type use mechanisms of ion exchange,
surface adsorption, partition, and size exclusion. Other chromatography
techniques are based on the stationary bed, including column, thin layer, and
paper chromatography.

Based on this approach three components form the basis of the chromatography
technique.
 Stationary phase: This phase is always composed of a “solid” phase or “a
layer of a liquid adsorbed on the surface a solid support”.
 Mobile phase: This phase is always composed of “liquid” or a “gaseous
component.”
 Separated molecules

The type of interaction between stationary phase, mobile phase, and

substances contained in the mixture is the basic component effective on
separation of molecules from each other.

The purpose of applying chromatography which is used as a method of

quantitative analysis apart from its separation, is to achieve a satisfactory
separation within a suitable time interval. Various chromatography methods have
been developed to that end. Some of them include column chromatography, thin-
layer chromatography (TLC), paper chromatography, gas chromatography, ion
exchange chromatography, gel permeation chromatography, high-pressure liquid
chromatography, and affinity chromatography.

Types of chromatography
a) Column chromatography - Since proteins have difference characteristic
features as size, shape, net charge, stationary phase used, and binding capacity,
each one of these characteristic components can be purified using
chromatographic methods. Among these methods, most frequently column
chromatography is applied. This technique is used for the purification of
biomolecules. On a column (stationary phase) firstly the sample to be separated,
 then wash buffer (mobile phase) are applied. Their flow through inside column
material placed on a fiberglass support is ensured. The samples are
accumulated at the bottom of the device in a time-, and volume-dependent
manner
b) Ion-exchange chromatography - Ion- exchange chromatography is based on
electrostatic interactions between charged protein groups, and solid support
material (matrix). Matrix has an ion load opposite to that of the protein to be
separated, and the affinity of the protein to the column is achieved with ionic ties.
Proteins are separated from the column either by changing pH, concentration of
ion salts or ionic strength of the buffer solution. Positively charged ion- exchange
matrices are called anion-exchange matrices, and adsorb negatively charged
proteins. While matrices bound with negatively charged groups are known as
cation-exchange matrices, and adsorb positively charged proteins.
c) Affinity chromatography - This chromatography technique is used for the
purification of enzymes, hormones, antibodies, nucleic acids, and specific
proteins. A ligand which can make a complex with specific protein (dextran,
polyacrylamide, cellulose etc.) binds the filling material of the column. The
specific protein which makes a complex with the ligand is attached to the solid
support (matrix), and retained in the column, while free proteins leave the
column. Then the bound protein leaves the column by means of changing its
ionic strength through alteration of pH or addition of a salt solution.
d) Paper chromatography - In paper chromatography support material consists of
a layer of cellulose highly saturated with water. In this method a thick filter paper
comprised the support, and water drops settled in its pores made up the
stationary “liquid phase.” Mobile phase consists of an appropriate fluid placed in
a developing tank. Paper chromatography is a “liquid-liquid” chromatography.
e) Thin-layer chromatography – With this method, lab technician use a silica gel
or alumina as its stationary phase and coat it onto a thin glass or plastic. The
mobile phase is like high performance liquid chromatography, in that it uses
either a singles solvent or a mixture of solvents.
Thin-Layer chromatography is best suited for smaller experiments that
require analyzing liquid or solvent compound, the number of components, or the
purity of compounds in a mixture.
f) Gas chromatography – This method is one of the most common in the
chromatography world. Like all other forms of chromatography. It has a mobile
phase and stationary phase. In this case, the mobile phase is an inert gas, and
the stationary phase is a column within the machine.
Gas chromatography is Ideal in any scenario that needs to separate
volatile mixtures.
g) High-pressure liquid chromatography (HPLC) – This method is favorite in
many labs because of its efficiency using a liquid (or solvent) mobile phase that
pushes the sample through the machine using a high – pressure pump. HPLC is
best for any scenarios that require separating non-volatile mixtures.

Initially chromatographic techniques were used to separate substances

based on their color as was the case with herbal pigments. With time its
application area was extended considerably. Nowadays, chromatography is
accepted as an extremely sensitive, and effective separation method. Column
chromatography is one of the useful separation, and determination methods.
Column chromatography is a protein purification method realized especially
based on one of the characteristic features of proteins. Besides, these methods
are used to control purity of a protein. HPLC technique which has many superior
features including especially its higher sensitivity, rapid turnover rate, its use as a
 quantitative method, can purify amino acids, proteins, nucleic acids,
hydrocarbons, carbohydrates, drugs, antibiotics, and steroids.

4. Neutron Activation Analysis (NAA)

Is an extremely sensitive technique used to determine the existence and
quantities of major, minor and trace elements in a material sample? NAA differs from
other methods in that it relies on the atom’s nucleus and ignores
chemical formulation, unlike mass-spectrometry or chromatographic methods. NAA
requires a source of neutrons, gamma-ray detectors and a thorough understanding
of how elements react to neutron bombardment.

NAA was discovered in 1936 by Hevesy and Levi, who found that samples
containing certain rare earth elements became highly radioactive after exposure to a
source of neutrons.
Forensic Analysis
Personnel of the NAA laboratory have considerable experience in the
forensic analysis of evidentiary materials. Bullet fragments, gunshot residue,
plastic, hair and fingernails, and geological materials are included among recent
examples. Comparing materials nondestructively is a chief advantage of NAA for
forensics.
High-Purity Materials
Materials such as high-purity silica, silicon, aluminum, other materials and
their compounds that do not form long-lived radionuclides, cellulose air filters, as
well as graphite are excellent matrices for high-sensitivity NAA. Such materials
can be irradiated in graphite rabbits for many hours in PT-1 for determinations of
many elements at the sub-ppb level. Silicon wafers and SiO2 used in fiber optics
are examples that have been analyzed.
Radiochemical separations:
Is primarily concerned with the study of radioactivity in naturally
occurring radioactive materials and in other materials in which radionuclides
and their compounds are produced by irradiation.
5. Electrophoresis - analysis is a method that is used to separate proteins of differing
sizes and/or charges. A sample of the protein, often DNA or RNA, is placed in wells
at one end of a gel. An electric current is applied that repels the proteins and causes
them to move down the gel. Because size and charge control how far the sample will
travel, the gel can then be used to determine the makeup of the protein. By
comparing the gel with the protein spread through it to a reference or standard
sample, an identification can be made. Electrophoresis analysis is used in forensics
to compare DNA, in medical laboratories to do genetic testing, and in microbiology
labs to identify microorganisms. In addition to analyzing proteins or DNA,
electrophoresis is also used to create purified samples of proteins. Once the sample
is run through the gel, the portion with the protein of interest can be removed and the
protein it contains can be used for analysis or testing.

6. Immunoassays –
An immunoassay is a type of bioanalytical test where detection of a target
compound (analyte) depends on a specific antibody-antigen reaction.
 Three main components: an Analyte, an Antibody, and a signal-generating label.
Immunoassays are popular test used in medical diagnostics and pharmacology
studies. They are also important in forensic science. This seminar will focus on two
forensic applications of immunoassays: their ability to detect trace amounts of drug in
money and fingerprints, and their ability to detect explosives residues.

Modern Technologies use for forensic science

Technology is quickly taking over every aspect of our lives, including solving
crimes. The rapid improvements in technology have meant that solving crimes almost
takes on a futuristic factor, like something from a work of fiction.
During the forensic science process, forensic equipment is used to process samples
and evidence to solve crimes. Measurements include analysis of evidence,
fingerprinting or DNA identification, analyzing drugs or chemicals, and dealing with body
fluids. Importantly, it’s the fusion of science and technology that allows forensic
scientists to do a lot of their work. Specifically, sciences such as biology, chemistry and
mathematics are combined with various technologies to process evidence.
There are many different technologies used in the forensic sciences that most people
don’t know exist. Some of these technologies include:

1. PHENOM DESKTOP SEM

The Phenom SEM is the best tool for forensic scientists as it offers a very simple
to use, fast and high-quality imaging tool with the added capability of determining
elemental composition. This assists scientists in analyzing evidence quickly and reliably
to help solve criminal cases. Specifically, the Phenom GSR Desktop SEM allows crime
labs to search for gunshot residue particles. It’s automated, and the software and
hardware are fully integrated, so it’s very user-friendly.
The Phenom GSR Desktop SEM also uses a unique system that combines automated
GSR analysis and classification software with element identification and the easy-to-use
Phenom XL desktop SEM. There are lots of software parameters, which can be
adjusted to individual needs. For example, you can optimize it for sensitivity or speed.

2. ALTERNATIVE LIGHT PHOTOGRAPHY

Alternative light photography is one of the quickest ways to detect whether
damage has been done to a body before it even surfaces on the skin. Used by forensic
nurses, alternative light photography can sometimes mean the difference between life
and death.
The camera uses blue light and orange filters to see whether bruising has occurred
below the skin’s surface, and ultraviolet light to enhance bruises, bite marks and search
for trace evidence. Infrared photography is used to enhance blood that’s difficult to see
on dark and patterned clothing and tattoos due to decomposition, lividity and burning.
3. DIGITAL SURVEILANCE FOR GAMING EQUIPMENT
Criminals sometimes hide illicit data on an Xbox in the hope that a gaming
console won’t be seen as a likely evidence target. With an XFT Device, once the Xbox
file system is mounted, an analyst can browse the directory tree, list its contents, open
and view files, and expand subdirectories and files. The XFT Device will give authorities
access to hidden files on Xbox hard drives, and can also record access sessions to be
used as evidence in court.

4. FACIAL RECONSTRUCTION
Facial reconstruction is a method used in the forensic field when a crime involves
unidentified remains. The process recreates the face of an individual from their skeletal
remains through a combination of artistry, anthropology, osteology and anatomy. There
are three main types of facial reconstruction: two-dimensional (photographic prints or
drawing), three-dimensional (sculpture or high-res 3D computer image) and
superimposition.

Whilst not always the most reliable technology, facial reconstruction is used by many
forensic labs to determine the appearance of victims who are too damaged, or whose
bodies are decomposing, to make a visual identification. The user inputs data into the
software and a possible physical appearance is deduced.

5. DNA SEQUENCING
DNA is used to identify both criminals and victims by using trace evidence, such
as hair or skin. DNA sequencing determines the order of the four chemical blocks
(called “bases”) that make up the DNA molecule. Although DNA evidence alone isn’t
enough to secure a conviction today, DNA profiling has become the gold standard in
forensic science since the first case more than 30 years ago.
DNA sequencing allows forensic scientists to sequence STR (short tandem repeat)
markers, potentially resulting in an increased ability to differentiate between individuals
in complex DNA mixtures. Additionally, alternate marker types such as SNPs (single
nucleotide polymorphisms) can be more easily integrated into casework laboratories,
resulting in new capabilities, such as ancestry or phenotype prediction in unsolved
cases.

6. AUTOMATED FINGERPRINT IDENTIFICATION

Fingerprint identification allows forensic scientists to compare fingerprints found
to an extensive digital database. Since fingerprint identification first emerged in the
1980s, automated fingerprint identification systems (AFIS) have become central to the
work of police and other law enforcement agencies around the world. By dramatically
increasing the potential for successful identification of a suspect, these systems have
fundamentally changed the way that authorities approach the investigation of a wide
range of criminal activities.
The rapid adoption of AFIS has inevitably led to further investment in developing it.
Enhancements include the introduction of palm prints, interfacing the AFIS with other
criminal justice information systems, interfacing with digital mugshots and livescan
devices, and the use of multi-modal biometrics. Newer technologies such as magnetic
fingerprinting dust also means investigators are able to get a perfect impression without
compromising the fingerprint. Condemn

7. LINK ANALYSIS SOFTWARE

Link analysis is a data analysis method in the network theory that examines the
connections or relationships between the network nodes. The connection or relationship
can be between any type of node or object like people, organizations, and transactions.
When it comes to funds, there’s often a mountain of paperwork. Link analysis
software is used by specialist accountants to highlight any strange financial activity
found within the paper trail. The software looks at financial transactions and the profile
of the customer, and using statistics it generates possible illegal behavior. It’s mostly
used by financial institutions like banks and insurance and cyber security agencies to
uncover criminal networks. In the same way, government agencies perform link analysis
to investigate frauds, improve screening processes, investigate criminal activities, and
expose the terrorist network.

9. FIRE TECHNOLOGY
Forensic scientists apply technologies to heat samples taken from the scene,
causing any residue to separate. This sample is then analysed to determine the
chemical structure. Scientists also use other tests such as using liquid nitrogen gas to
trap residue which is then analysed using gas chromatography. The investigation will
include closely surveying the damaged scene to establish the cause of the fire.

11. 3D SCANNER
A 3D scanner is generally one of the most expensive but brilliant pieces of
equipment available to crime scene personnel. You can take the scanner, place it in the
middle of a room, and turn it on. The scanner will then rotate 360 degrees while taking
photographs of the entire room and the objects in it. It will also measure distances from
the scanner to the walls, and from the scanner to objects in the room.
Before the 3D scanner, drawing a room was time-consuming and labor-intensive. It also
required a great deal of artistry skills. Items were often left out to avoid ‘cluttering’ a
drawing, which resulted in an inaccurate account of the room. A 3D scanner gives
accuracy in 10-30 minutes, which is significantly less time than it would take an artist.
Classes/Types of Evidence –
Evidence is anything that can be used to determine whether a crime has been
committed. Evidence may link a suspect to a scene, corroborate or refute an alibi or
statement, identify a perpetrator or victim, exonerate the innocent, induce a
confession, or direct further investigation.

Direct and circumstantial evidence

Direct evidence establishes a fact. Examples of direct evidence are eyewitness

statements and confessions. Circumstantial evidence, on the other hand,
requires that a judge and/or jury make an indirect judgment, or inference, about
what happened.

Physical versus biological evidence

Forensic evidence can be divided into two basic categories: physical and
biological. Physical evidence may take the form of nonliving or inorganic items,
such as fingerprints, shoe and tire impressions, tool marks, fibers, paint, glass,
drugs, firearms, bullets and shell casings, documents, explosives, and petroleum
byproducts or distilled fire accelerants. Biological evidence, on the other hand,
includes organic things like blood, saliva, urine, semen, hair, and botanical
materials, such as wood, plants, pollens and yes, Clarice, moth cocoons.

Reconstructive evidence

Any evidence that helps law enforcement officer better grasp what happened at
the crime scene is considered reconstructive evidence. Broken glass or pried-
open doors and windows often reveal a perpetrator’s points of entry and exit, and
determining whether a window was broken from the inside or the outside tells
which way the perpetrator went through it.
Evidence derived from shoeprints, blood spatters, or the trajectory of bullets may
pinpoint where in the room everyone was located and exactly how and in what
sequence the events of the crime occurred. Whether the victim was attacked
from the front or from behind, whether the life was taken quickly or after a
struggle, and whether the prime suspect was at the scene at the time of the
murder are important aspects in creating a clearer picture of the crime scene.

Some Evidence Types and Applicable Instrumentation Methods Categories and

Types of Physical Evidence.

Classes/Types of Evidence
Physical Chemical Biological
Plastics (Pieces) Drugs and Toxic Blood
Substance
Glass (pieces) Paints, pigments Body fluids
Imprints (Fingerprints) Gunshot residues Hair
Indentations (tire Volatile substances Tissues
 impressions)
Striations (riffing Accelerants, solvents Pollen
impressions on bullets) alcohols (esp. ethanol)
Physical patterns Rubber materials Wood materials other plant
derived matter
Firearms, bullets, cartridge Resins, plastic Feathers
cases
Tool marks Explosive residues
Questioned Documents Fibers
Soil, Glass and
Miscellaneous trace
evidence

Some Evidences Types and Applicable Instrumental Method

Evidence type Applicable Instrumental method
Blood, Body Fluid, Tissue Spectrophotometry, Enzyme Assays
Blood, Body Fluid, Tissue genetic marker Electrophoresis, Isoelectric Focusing
analysis
Drugs, Toxic Substances UV and IR Spectrophotometry, X-ray
Diffractometry, Microscopy, GC/MS, HPLC
Paints, Pigments IR and Atomic Absorption
Spectrophotometry, Polarized Light and
Scanning Electron Microscopy, Emission
Spectrometry
Explosive Residues IR Spectrophotometry, Polarized Light
Microscopy, HPLC
Gunshot Residues Neutron Activation Analysis, Atomic
Absorption Spectrophotometry, SEM
Accelerate, Solvents Gas Chromatography
Ethyl Alcohol (Blood or Breath) Gas Chromatography, Enzyme Couple
Assay
Hair Microsocpy, Electrophoresis, Isoelectric
Focusing
Fibers Polarized Light Microscopy, IR and Visible
Spectrophotometry, Micro
spectrophotometry (UV, VIS, IR)
Instrumental Method or Methodology Categories and Types of evidence to
which applicable
Microscopically Visible General Examination; Biological; Question
Documents
Polarized Light Crystal, Mineral, Soil, Paints, Glass,
Explosive Residue
Scanning Electron (SEM) Gunshot Residue, Paints, Microphysical
Matching
Spectrophotometer/Spectral Drugs, Biochemical assay, Enzyme Assay,
Ultraviolet/Visible Organic Dyes and Pigments, Inks
Infrared Drugs, Fibers, Paints, Oils, Plastics,
Organic
Fluorometry Oils, Fluorescent organic substance,
Gunshot Residue
Atomic Absorption Gunshot Residue, Quantitative Elemental
 Analysis,
Atomic Emission Alloys, Paints, Qualitative Elemental
analysis
X-ray Diffractometry Drugs, Crystal, Minerals, Alloys, Metal
Mass Spectrometry (inc. GC/MS) Organic substance, Drugs, Accelerants,
Mixtures of Organics
Neutron Activation Analysis Gunshot Residue, High Sensitivity
Elemental Analysis
Chromatographic/Separation Thin – Drugs, Inks, Pigment, Pollutants Volatiles,
Layer (TCL), Gas Chromatography (GC) solvents, Alcohols, Toxins, Accelerants

Advantages
There are various advantages of forensics. It has a broad spectrum of applications
which are very useful to us. Some of the advantages of this science are discussed
below.

☛ With the help of certain computer tools, it is possible to control cybercrime. This is
done through packet sniffing (sensing critical information in the data packets), IP
address tracing (to get the address from where the criminal was accessing), email
address tracing (to get the details of the email server and in cases of email bombs).
This is called computer forensics.
☛ It helps in determining the cause of death by examining the postmortem changes,
blunt injuries, burns and scalds on the body, and the scene of death. If it’s sudden
natural death, the case is investigated by the coroner or a medical examiner.
☛ Forensic analysis is used to investigate accident cases and to determine its cause by
analyzing the vehicle condition, tire and other marks, eye witnesses, calculating the
vehicle’s speed etc.
☛ The alcohol content in a human being can be determined by analyzing the blood and
other body fluids like saliva, urine etc.
☛ It also includes anthropology and helps in sex determination.

☛ Clinical forensic medicine is useful in finding out child abuse, defensive wounds on a
victim, gunshot wounds, injury patterns in domestic violence victims, self-inflicted
injuries, sexual assault, and semen persistence.
☛ Biometrics technology is combined with forensics, which helps to identify the
fingerprint of the criminal, on the objects present in the crime scene.
☛ Phonetics, which is also a part of forensics that is used to tap the voice signals and
identify the speaker. Speech enhancement, speech coding and tape authentication are
other techniques used in phonetics.
☛ Other useful aspects of forensic analysis includes fire investigation, forgery and fraud
in payment cards, lie detection, footprint marks, voice analysis, digital imaging and
photography etc.

Disadvantages
Despite numerous advantages of this science, there are some ethical, legal, and
knowledge constraints involved in forensic analysis.
☛ DNA analysis of a person is believed to be against human ethics, as it reveals private
information about an individual.
☛ Equipment used in forensics is expensive.
☛ Scientific analysis consumes lot of time because of which the verdict is delayed.
☛ It requires precise and accurate analysis. Even if a minor error occurs in the analysis,
it may result in the wrong figure.
☛ The evidence cannot be accessible at all times.
☛ Evidence is prone to manipulation, which may end up in an unrighteous verdict.
☛ Interpretation of the analysis differs from one forensic scientist to another.
☛ Forensic analysis can be prevented by strong influences (political or financial factors).
☛ There is no particular standard to verify the result of the experiment. It requires wide
knowledge and intensive study.
☛ Innovation is hindered as the approach is mostly the same.
☛ Misconceptions and ignorance can mislead the experimental analysis.
☛ Maintaining privacy and secrecy of the information gathered through forensic analysis
is quite difficult.