Forensic microscopy

Article by:
Jones, Edwin L., Jr. Forensics Department, Ventura County Sheriff's Crime Laboratory, Ventura, California.
Publication year:2014
DOI:http://dx.doi.org/10.1036/1097-8542.801800
Content
 Stereomicroscope
 Biological microscope
 Polarized light microscope
 Comparison microscope
 Scanning electron microscope
 Digital forensic microscopy
 Bibliography
 Additional Readings

The application of microscopy techniques for purposes of civil or criminal law. Edmond
Locard (1877–1966) stated that every contact leaves a trace. This is known as the
Locard exchange principle, the basis for much of forensic microscopy. Since the early
1800s, the microscope has been used to help solve crimes. Today, it remains one of the
most used tools in the crime laboratory. Criminal evidence ranging in scale from
micrometer-sized particles to hair and paint chips can be found and removed to a
laboratory for microscopic analysis, where its history may be deduced to help solve the
crime. Microscopy can provide insight into the identity and origin of a material, what has
happened to it and when, and the routes it may have taken between a crime victim,
suspect, and crime scene (Fig. 1). See also: Criminalistics

SAVE
Fig. 1 Hypothetical flow of trace evidence between a crime scene, victim, and suspect. Seven types of trace
evidence were transferred from the crime scene to both the suspect and victim. Three types of trace
evidence were found at both the crime scene and on the victim. One type of trace evidence was transferred
from the victim to the suspect and the crime scene. One type of evidence was transferred from the victim to
the suspect. One type of evidence was transferred from the suspect to the crime scene.

Stereomicroscope
The low-powered stereomicroscope is probably the most used microscope in the crime
lab. The magnification typically ranges from 7× to 40×, and the image is upright (not
reversed) and three-dimensional. With additional lenses, the magnification can range
from below 3× to above 100×. This type of microscope can be used with a variety of
lighting techniques such as transmitted, fluorescence, diffuse, coaxial, and oblique
illumination. See also: Fluorescence microscope; Microscope; Optical microscope

The stereomicroscope is used in drug analysis to help identify marijuana and to

evaluate powders and chunks of material that may need to be separated before
analysis. In examining firearms and tool marks, it is used to evaluate striations or
imprint marks, look for trace evidence, evaluate bullet holes for muzzle blast, and
search for unburned gunpowder particles. In addition, when a bullet passes through an
object, some of that object may be transferred to the bullet and detected
microscopically. In forensic biology, the stereomicroscope is used to find and measure
small bloodstains, to evaluate a bloodstain to determine if it came from the inside or
outside of a garment, and to remove stains from hidden places. When examining
documents, it can be used to evaluate typefaces, examine strikeovers and ink lines that
cross over, and find erasures or alterations. For trace evidence, the stereomicroscope is
the tool of choice for finding and characterizing hairs, fibers, paint, glass, soil, and
building material. See also: Forensic biology

Biological microscope
The biological microscope is used for examining biological fluids, human tissue, hairs,
fibers, drug crystals, food, and other stains. A typical biological microscope has 4×,
10×, 20×, and 40× objectives on a revolving nosepiece. It is normally equipped with
10× eyepieces giving a final magnification of from 40× to 400×. Many additional lenses
can be added to this style of microscope to increase the range of magnification from
less than 20× to greater than 1500×. In addition, these microscopes can be modified to
include darkfield, phase contrast, fluorescence, dispersion staining, darkfield epi-
illumination, and reflected coaxial illumination. When darkfield epi-illumination or
reflected coaxial illumination is used, the biological microscope is transformed into a
metallurgical microscope, which can be used to examine surface detail and opaque
objects. See also: Phase-contrast microscope; Reflecting microscope

A forensic pathologist typically uses the biological microscope to examine tissue

samples from injuries to determine if the injuries occurred before, during, or after death.
In the forensic biology section of the laboratory, the microscope is typically used to
identify sperm in rape cases. This process involves differential staining and examining
the slide at greater than 200×. When sperm are identified from samples collected from
a victim, this is proof that sexual activity has taken place. The legal issue of consent is
not addressed by the presence of sperm. Other body fluids such as vaginal secretions,
saliva, and feces might have been mixed with semen, and microscopy can give vital
clues as to the origins of these other body fluids.

Some crime labs use crystal tests to identify drugs such as phencyclidine,
amphetamines, cocaine, barbiturates, heroin, morphine, and codeine. In a typical
crystal test, a small amount of the drug and a liquid reagent are placed on a microscope
slide. After a short time, unique crystals for that drug can be recognized at 100× with a
biological microscope.

Polarized light microscope

The polarized light microscope is a biological microscope with special modifications,
including a polarizer, analyzer, rotating stage, accessory slot, Bertrand lens, and a flip-
up condenser. This microscope is typically used to identify synthetic fibers, minerals,
glass, starch, and many other types of particles. It has the advantage of measuring
optical properties without altering the sample. Six major synthetic fiber types (acrylic,
acetate, rayon, olefin, nylon, and polyester) can be quickly differentiated with the
polarized light microscope (Fig. 2). A significant amount of training and experience, as
well as reference standards, is necessary to master the use of this instrument, since
most universities do not teach this subject. The McCrone Research Institute in Chicago
has trained many forensic scientists in the use of the polarized light microscope. See
also: Birefringence; Polarized light microscope

SAVE
Fig. 2 Crossed polarizers showing interference colors (represented here in black and white) of three
colorless synthetic fibers. These interference colors can be used to calculate the birefringence, which is an
optical property used to identify these fibers. The birefringence of polypropylene is medium, nylon is high,
and polyester is very high.

Comparison microscope
A comparison microscope consists of two identically equipped compound microscopes
joined together with an optical bridge, giving a split-screen image. The comparison
microscope is used to compare bullets, cartridge cases, and toolmarks. This microscope
normally uses low-powered objectives and reflected, diffused, or oblique illumination.
The comparison microscope is also used to compare trace evidence such as hairs and
fibers. It uses the same range of magnifications as the biological microscope.

One such case illustrating the comparison of hair, as well as DNA analysis, involved an
elderly male victim who was stabbed in a robbery at a hamburger stand. Clumps of long
dark hair that appeared to have been pulled out during a struggle were found in several
locations at the crime scene. Microscopic examination of that hair showed more than 50
anagen roots. Anagen roots are those still attached to the scalp with a blood vessel and
a nerve, indicating that the clumps of hair were forcibly removed during the struggle.
The hair from the crime scene had a maximum length of 17 in. The maximum length of
the victim's gray hair was less than 1 in. The police collected and the crime lab
compared hair from several long-haired suspects with negative results. Finally, a female
drug user was identified by a relative as the murderer. A hair sample was collected and
submitted to the hair examiner, who testified that the hairs from the crime scene
probably came from the suspect. A blood sample from the suspect along with 15 roots
from the crime scene hairs were submitted to a DNA laboratory (Fig. 3). The DNA
matched, and the suspect was convicted. This case became the first California DNA
case to successfully get through the Appeals Court.

SAVE
Fig. 3 Fifteen anagen hair roots sent for DNA typing.

Scanning electron microscope

The scanning electron microscope with energy dispersive x-ray spectrometer (SEM/EDX)
is routinely used to identify the elemental content of very small samples. The most
common sample analyzed with the SEM/EDX is gunshot residue. This is done by dabbing
the shooter's hands with sticky tape attached to an SEM stub (a substrate for mounting
specimens). The stub is then examined for gunshot residue (GSR) particles (usually 1–6
micrometers) that contain lead, barium, and antimony. Many of the SEM/EDX
microscopes are connected to computers that have programs that can search
automatically for GSR particles. See also: Scanning electron microscope

Digital forensic microscopy

Most microscope systems can be modified to capture digital images, which can be sent
by e-mail to forensic scientists for examination and comment. An automated system for
glass refractive index measurement uses a phase-contrast microscope, hot stage, video
system, and computer to compare glass samples. The National Integrated Ballistics
Identification Network (NIBIN), sponsored by the Federal Bureau of Investigation (FBI)
and Bureau of Alcohol, Tobacco and Firearms (ATF), uses digitally captured images
(through the microscope) of cartridge cases or bullets to link shooting incidents with
guns taken from suspects in distant jurisdictions.
 The infrared spectrophotometer with a microscope attached is a common tool in crime
laboratories. It is used to perform spectrophotometric analysis on very small samples
such as paints, plastics, fibers, and many other substances. The
microspectrophotometer is used to measure the spectrum of the visible colors,
ultraviolet radiation, and fluorescence that are present in microscopic objects. An
automated version of the microspectrophotometer using a remote control stage and
computer can search tape lifts taken from suspects, victims, and crime scenes. First a
fiber is chosen and entered into the apparatus. The tape lifts are then searched for that
target fiber.

Another digital system is a portable microscope with fiber-optic cables that is used to
acquire high-quality microscopic images such as a paint transfer on a bumper or hood
of a vehicle without dismantling.

Edwin L. Jones, Jr.

Bibliography
 P. De Forest, Foundations of forensic microscopy, in R. Saferstein (ed.), Forensic Science
Handbook, Prentice Hall, Upper Saddle River, NJ, 2002
 B. Fisher, Techniques of Crime Scene Investigation, CRC Press, Boca Raton, 1993
 J. Houde, Crime Lab: A Guide for Nonscientists, Calico Press, Ventura, 1999
 S. Palinik, Microscopy, in J. A. Siegal et al. (eds.), Encyclopedia of Forensic Sciences,
Academic Press, San Diego, 2000
 R. Saferstein, Criminalistics: An Introduction to Forensic Science, Prentice Hall, Upper
Saddle River, NJ, 2001

Additional Readings
 R. M. Brown and J. S. Davenport, Forensic Science: Advanced Investigations, South-
Western, Mason, OH, 2012
 M. M. Houck and J. A. Siegel, Fundamentals of Forensic Science, 2d ed., Academic Press,
Burlington, MA, 2010