J O U R N A L O F

CLINICAL
FORENSIC
MEDICINE
Journal of Clinical Forensic Medicine 13 (2006) 215218
www.elsevier.com/locate/jcfm

Short report

The use of an alternative light source to detect semen in clinical

forensic medical practice q
a,*
C.A. Lincoln , P.M. McBride b, G.R. Turbett c, C.D. Garbin c, E.J. MacDonald d

a
Clinical Forensic Medicine Unit, Gold Coast Oce, Level 2, Surfers Paradise Police Centre, 68 Ferny Avenue, Surfers Paradise Qld 4217,
South East Queensland, Australia
b
Sexual Assault Medical Ocer, WA, Australia
c
Managing Scientist, Forensic Biology, PathWest Laboratory Medicine, WA, Australia
d
Clinical Leader, Wellington Sexual Health Service, 17 Adelaide Rd, Newtown, Wellington, New Zealand

Available online 31 March 2006

Abstract

One of the primary aims of forensic examination in sexual oences is to detect and recover biological material that will link the oen-
der with the complainant. One potentially valuable method by which trace biological evidence may be identied in other forensic settings
is via the use of an Alternate Light Source (ALS).
The aim of this study was to determine whether or not there was any potential benet in using an ALS as an adjunct in sexual assault
examinations to aid the detection of forensically relevant areas on the body which are not identiable on visual inspection for sampling.
We present two case reports, which illustrate the potential value of using an ALS in clinical forensic medical practice as an adjunct in
sexual assault examinations to detect potentially forensically useful areas of skin to sample for semen.
Prior to introducing the ALS into our clinical forensic medical practice, we undertook a number of simple laboratory studies to deter-
mine a protocol for its use. Semen is known to uoresce using an ALS at a wavelength of 450 nm. Although we did not conduct a rigorous
scientic evaluation of the technique, we evaluated the use of an ALS to detect semen on a range of inanimate surfaces as well as human
skin. On all surfaces, visibility of uorescence was increased by reduced distance of light source from the surface and increased concen-
tration of semen on the surface, but was not noticeably aected by the angle at which the light source was held in relation to the surface.
Crown Copyright 2006 Published by Elsevier Ltd and AFP. All rights reserved.

Keywords: Semen; Fluorescence; Alternative light source

1. Introduction of an ALS to detect recently deposited semen and urine on

An Alternative Light Source (ALS) equipped with exci-
tation lters has been used successfully to detect nger-
prints, body uids and trace evidence in a number of
studies.1,5 These studies involved uorescence of biological
uids and stains on inert materials. Neither provided
guidelines for the use of an ALS in the clinical setting or
demonstrated the benet of detection of biological material
on human skin. More recently, a study comparing the use
q
This paper is part of the special issue entitled Sexual Oences, guest
edited by Dr. Guy Norfolk and Dr. Cath White.
*
Corresponding author. Tel.: +617 5570 7829; fax: +617 5570 7800.
E-mail address: Lincoln.CatherineA@police.qld.gov.au (C.A. Lin-
coln).
skin and cloth has cast doubt on the benet of an ALS in
clinical practice.3 The authors of this paper also provide a
comprehensive explanation of various light sources avail-
able for use in the forensic setting and in particular, refer
to another recent paper casting doubt on the ecacy of a
Woods Lamp in uorescing semen.4
One of the primary aims in the forensic examination of
subjects in sexual oence investigations is to detect and
sample biological material with the potential to link them;
thus any means of facilitating or enhancing the detection of
forensically useful areas on the body from which to sample,
could be of signicant value.
The rst part of this report addresses some practical
aspects of the use of an ALS on both inanimate surfaces

1353-1131/$ - see front matter. Crown Copyright 2006 Published by Elsevier Ltd and AFP. All rights reserved.
doi:10.1016/j.jcfm.2006.02.016
216 C.A. Lincoln et al. / Journal of Clinical Forensic Medicine 13 (2006) 215218

and on human skin. The second part discusses two clinical  ALS held at 20 cm from surface at an angle of 90;
cases in which the use of an ALS to direct the collection of  ALS held at 10 cm from surface at an angle of 90;
forensic samples from the body of a sexual assault com-  ALS held at 5 cm from surface at an angle of 90;
plainant was found to be forensically valuable.  ALS held at 5 cm from surface at an angle of 45.

2. Equipment and materials The uorescent areas were rated by two reviewers who
took into account both the brightness and contrast to give
Alternative light source: PoliRay (Ron Australia Pty a subjective measure of the overall visibility of the uores-
Ltd., Dingley, Vic., Australia), with an excitation lter of cent areas.
450 nm and barrier lter goggles. Visibility of observed uorescent areas were rated by
Pentax MZ-50 SLR camera, 2880 mm macro lens and scaling a combination of the brightness and contrast from
KV 550 nm camera lter. 0 to ++++ and recorded.
Semen from a fertile non-vasectomised male.
Surfaces examined: 3.2. Human skin (anterior forearm)

 Green cotton material; The forearm of a volunteer was examined by two inde-
 Light blue denim material; pendent observers using the ALS to ascertain whether there
 Plastic transparency lm; were any areas of uorescence present prior to the applica-
 White absorbent paper; tion of semen.
 Human skin anterior forearm. Three areas of semen at volumes of 5 ll, 10 ll
and 20 ll were then applied to the forearm of the volun-
teer, allowed to dry and examined by two independent
3. Method observers using barrier lter goggles under the following
conditions:
3.1. Inanimate surfaces
 ALS held at 20 cm from surface at an angle of 90;
Each of the four types of inanimate surface was stained  ALS held at 10 cm from surface at an angle of 90;
with 100 ll of semen, so that the semen stain covered a cir-  ALS held at 5 cm from surface at an angle of 90.
cular area of approximately 1 cm diameter.
Each surface was then examined by three independent Visibility of observed uorescent areas were rated by scal-
observers using barrier lter goggles under the following ing a combination of the brightness and contrast from 0 to
conditions: ++++ and recorded (see Fig. 1).

Fig. 1. Semen at volumes of 5 ll, 10 ll and 20 ll on anterior forearm skin (1 cm diameter disc) viewed through barrier lter with alternative light source at
wavelength 450 nm held at 20 cm.
 C.A. Lincoln et al. / Journal of Clinical Forensic Medicine 13 (2006) 215218
4. Results
On the inanimate surfaces, all observers found that vis-
ibility of the uorescence was increased by reduced distance
of light source from surface and increased concentration of
semen on the surface, but not noticeably aected by the
angle at which the light source was held in relation to the
surface.
On human skin, both observers found that visibility of
the uorescence was increased by reduced distance of light
source from surface and increased concentration of the
semen.
Prior to the application of semen to the forearm, the
forearm of the volunteer was examined for areas of uores-
cence. Several uorescent areas were noted on the skin as
follows:
(i) dried skin around ngernails;
(ii) thickened skin or calluses on palms;
(iii) diuse areas thought to be hand-cream.
The uorescence observed in (i) and (ii) correlated with
areas of skin that were identiable on visual inspection.
The uorescence observed in (iii) was indistinct, mottled
and widespread and did not correlate with any distinct fea-
ture identiable on visual inspection.
5. Case report 1
A 28-year-old woman alleged that a male had rubbed his
penis between her labia and ejaculated. She had not washed
or showered since the incident and was forensically exam-
ined 4 h afterwards. On macroscopic visual examination,
no dried uid was noted on the external genitalia. The
ALS was then used to examine the external genitalia, and
several areas of uorescence were noted; in particular,
two areas on the inner aspect of the labia majora. A swab
was taken of each of these areas. The areas of uorescence
were noted to disappear after swabbing. In addition to
these ALS-directed swabs, routine genital swabs were col-
lected from the inner thighs, external and internal aspects
of the labia, lower vagina, and the perineum without the
use of the ALS. Laboratory analysis of swabs taken with-
out use of the ALS revealed very occasional sperm heads
on four of the swabs and no sperm on the others. Analysis
of the two ALS-directed swabs revealed a number of com-
plete sperm and clumps of multiple sperm heads on one
swab and no sperm on the other. The positive ALS-direc-
ted swab was used for DNA extraction and proling.
6. Case report 2
A 25-year-old woman who alleged attempted penile vag-
inal penetration with ejaculation was forensically examined
within 12 h of the alleged incident. She said that she had
been menstruating at the time of the alleged incident, and
had not washed or showered since. On visual examination
217
in normal light, there were extensive areas of dried blood
on the inner aspect of both upper thighs. With the use of
the ALS, multiple small deposits of uorescent material
were noted and collected with swabs. The areas of uores-
cence were noted to disappear after swabbing. Routine
inner thigh swabs2 were then collected without the use of
the ALS, using the double-swab technique.6 Laboratory
analysis of swabs taken without using the ALS revealed
four sperm heads on the left inner thigh swab and two
sperm heads on the right inner thigh swab. Analysis of
the inner thigh swabs taken from the areas of uorescence
identied with the ALS revealed multiple sperm heads on
the left thigh swab and multiple sperm heads on the right
thigh swab. The ALS-directed swabs were used for DNA
extraction and proling.
7. Discussion
Laboratory experimentation with the ALS revealed that
on all surfaces including human skin, the subjective visibil-
ity of uorescence was increased by reduced distance of
light source from the target surface and also with an
increased concentration of semen, but was not noticeably
aected by the angle at which the light source was held in
relation to the surface.
A number of false positives were noted using the ALS in
the laboratory setting to detect uorescence on human
skin; two of these were easily identiable with the naked
eye as areas of thickened or dried skin and one was thought
to be hand-cream.
One of the uorescent areas swabbed in the rst case
study did not yield sperm, yet had a similar appearance
to the other uorescent area swabbed which yielded multi-
ple sperm heads. This implies that other material in the
genital area may uoresce and be indistinguishable from
semen. It is worth remembering that the ALS is not
semen-specic, and that, in principle, any dried biological
material may uoresce.
The fact that the ALS was used to direct swabbing prior
to the collection of routine genital swabs may have reduced
the yield of semen from the routine non-ALS directed
swabs taken afterwards, as the uorescent areas could be
said to have been within the general area that would be
swabbed routinely. However, we do not consider this to
negate their potential value. The examination protocol
was decided upon in an eort to avoid the partial removal
of material by taking a routine swab and leaving a less con-
centrated deposit of semen which may not have been
detected by the ALS, given that higher concentrations of
semen are more likely to uoresce. A larger amount of
semen present on a single swab is more likely to yield sperm
material during laboratory analysis. In addition, the wiping
of larger areas of skin instead of smaller dened areas to
collect semen increases the likelihood of simultaneously
collecting epithelial cells of the subject in addition to
semen, thereby reducing the purity of the sperm material
on the swab.
218 C.A. Lincoln et al. / Journal of Clinical Forensic Medicine 13 (2006) 215218
Clinician variability in both the extent of swab contact
with the skin of the external genitalia and the pressure used
during routine forensic swabbing should also be acknowl-
edged. In particular, forensic sampling from all surfaces
of the external genitalia, especially the labia as in Case
Report 1, can be dicult even when special care is taken.
When sampling from areas of skin which are larger, such
as the inner thigh as in Case Report 2, routine swabbing
with a cotton swab will not necessarily make contact with
all inner thigh skin, even with the best intentions of the cli-
nician. The pressure used when placing the swab onto the
skin and thus the likelihood of collecting material, may
also dier when swabbing a larger area blindly as opposed
to swabbing a smaller dened uorescent area.
In Section 6, the dried blood was clearly visible in nor-
mal light and did not interfere with identication of uo-
rescent areas using the ALS, because the blood appeared
dark at the light wavelength used (450 nm).
While the denitive nature of any areas of uorescence
cannot be determined on visual inspection and will always
require laboratory analysis for identication, an ALS may
usefully direct the forensic medical examiner to areas of
particular interest for forensic sampling. In the case of a
complainant who gives a history of suspected ejaculation,
but where semen cannot easily be visualised upon initial
examination, an ALS may optimise collection of forensi-
cally useful samples. This could be especially useful in cases
where allegations involve ejaculation onto parts of the
body other than the genital area. Conning swab collection
to the blind swabbing of large areas of skin such as the
abdomen or chest where a complainant thinks ejaculate
may have landed is less likely to yield useful amounts of
material than directed sampling.
Both case reports described reveal the potential benet
of using an ALS as an adjunct to the routine blind foren-
sic samples taken as part of most sexual assault forensic
examination protocols. It should be noted that the authors
do not advocate use of the ALS instead of the routine
forensic sampling protocol, but suggest that it may be use-
ful as a screening tool prior to the collection of routine
swabs for the reason outlined above. Further investigation
into the use of the ALS in clinical forensic medical practice
is required to identify other materials likely to uoresce.
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