ORIGINAL ARTICLE
Forensic Image Analyses of Skin and Underlying Muscles as a
Tool for Postmortem Interval Delimitation
Histopathologic Examination
El-Shaymaa El-Nahass, DVM, PhD, Walaa A. Moselhy, DVM, PhD, and Nour El-Houda Y. Hassan, DVM, PhD
Abstract: One of the biggest challenges for forensic pathologists is to
diagnose the postmortem interval (PMI) delimitation; therefore, the
aim of this study was to use a routine histopathologic examination and
quantitative analysis to obtain an accurate diagnosis of PMI. The current
study was done by using 24 adult male albino rats divided into 8 groups
based on the scarification schedule (0, 8, 16, 24, 32, 40, 48, and 72 hours
PMI). Skin specimens were collected and subjected to a routine histo-
pathologic processing. Examination of hematoxylin-eosinstained sections
from the skin, its appendages and underlying muscles were carried out. Mor-
phometric analysis of epidermal nuclear chromatin intensities and area
percentages, reticular dermis integrated density, and sebaceous gland
nuclei areas and chromatin condensation was done. Progressive histo-
pathologic changes could be detected in epidermis, dermis, hypodermis,
underlying muscles including nerve endings, and red blood cells in relation
to hours PMI. Significant difference was found in epidermal nuclear
chromatin intensities at different-hours PMI (at P < 0.001). The highest
intensity was detected 40 hours PMI. Quantitative analysis of measurements
of dermal collagen area percentages revealed a high significant difference
between 0 hours PMI and 24 to 72 hours PMI (P < 0.001). As the PMI
increases, sebaceous gland nuclei and nuclear chromatin condensation
showed a dramatic decrease. Significant differences of sebaceous gland
nuclei areas between 0 hours and different-hours PMI (P < 0.001) were
obtained. A combination between routine histopathologic examination
and quantitative and morphometric analysis of the skin could be used to
evaluate the time of death in different-hours PMI.
Key Words: histopathology, imageJ, postmortem interval delimitation,
skin
(Am J Forensic Med Pathol 2017;00: 0000)
T he determination of the time of death is one of the most
important long-standing problems in the field of forensic
medicine.1 Postmortem interval (PMI) is defined as the time
elapsed between the death and autopsy.2 An accurate estimation
of the PMI is an important task.3 Several techniques have been
used to determine the PMI; therefore, development and enhance-
ment of those techniques are greatly needed in forensic medicine
in order to enable objective and quantitative analyses of forensic
images.4 Computerized image analysis technique is a useful
and promising tool for the estimation of PMI with a good objec-
tivity and reproducibility as a quantitative indicator within the first
36 hours following death in rats.5 Meanwhile, the usage of such
technique may facilitate the detection of inconsistencies that result
from human visual system examination.4
Manuscript received November 2, 2016; accepted January 22, 2017.
From the Faculty of Veterinary Medicine, Beni-Suef University, Beni-Suef, Egypt.
The authors report no conflict of interest.
Reprints: El-Shaymaa El-Nahass, DVM, PhD, Faculty of Veterinary Medicine,
Beni-Suef University, Beni-Suef 62511, Egypt. E-mail: shima_k81@yahoo.com.
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ISSN: 0195-7910/17/00000000
DOI: 10.1097/PAF.0000000000000301
Am J Forensic Med Pathol Volume 00, Number 00, Month 2017
Copyright 2017 Wolters Kluwer Health, Inc. Unauthorized reproduction of this article is prohibited.
Histological examination of the skin/appendages is tradition-
ally done by few researchers. However, few attempts had been
made to analyze structural skin changes and appendages simulta-
neously and to compare obtained data with PMI.68
Application of computational morphometry in histological
cuts plays an essential role in both medical and biomedical
research and increases the objectiveness and reproducibility.912
Availability of specific free software such as ImageJ allows
the opportunity for quantifying the research in many fields
including dermatology.11,13,14
The current work applied an easy, simple, and highly infor-
mative method using ImageJ software on quantification of post-
mortem changes of routinely stained skin specimens. Therefore,
the aim of the present study was to evaluate different PMI
delimitations using (a) histopathologic examination of skin, its ap-
pendages, and underlying muscles and (b) morphometric exami-
nation of the epidermal and dermal layers of skin by using
computerized image analysis techniques.
MATERIALS AND METHODS
Animals and Experimental Design
Twenty-four adult male albino rats weighing 150.55 8.56 g
were obtained from the Breeding Unit of the Egyptian Organization
for the Biological and Vaccine Production and were kept under
standard laboratory conditions, fed with a commercial diet (con-
taining 24% protein), and given water ad libitum. The procedure
was carried out according to the Experimental Animal Ethics
Committee, Faculty of Science, Beni-Suef University. Rats were
killed via cervical dislocation and then classified into 8 equal
groups according to the scarification schedule based on collecting
dorsal skin samples at 0, 8, 16, 24, 32, 40, 48, and 72 hours PMI.
Samples were carefully taken to avoid traumatic artifact. Each
specimen was fixed in 10% neutral-buffered formalin for routine
histopathologic examination. Preservation of dead rats was done
in an indoor, shaded, and cool temperate climate (average temper-
ature 15C).
Morphometry
Skin sections were examined and captured using a digital
camera (DM2500 M; Leica, Germany), with Adobe Photoshop
CS3 (Middle eastern version 10; Adobe Systems Inc., San Jose,
CA) and a freeware version of ImageJ 1.51d downloaded from the
National Institutes of Health website (http://rsb.info.nih.gov/ij).15
Parameters chosen for morphometric analysis were area, area
fraction, minimum and maximum gray values, perimeter, and
integrated density. Those parameters were assessed inside a
standard measuring frame of a known area using 50 captured
images each slide at oil immersion lens (1000) and routinely
stained with hematoxylin-eosin (HE). Prior to measuring, the
scale was changed to micrometers using set scale analysis.
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El-Nahass et al Am J Forensic Med Pathol Volume 00, Number 00, Month 2017

Nuclear Chromatin Intensities and Area Percentages
of Epidermis
Specified areas of epidermal layer were cropped to dimension
of 1  6 inches using crop tool in Adobe Photoshop CS3.16 Chro-
matin volume calculations were processed in ImageJ. Captured
images were corrected by enhance contrast (at level of 0.2). For
the calculation of chromatin volumes, the HSB (hue, saturation,
and brightness) stack splitter was used in order to converts RGB
colored images to 3-slice (hue, saturation, and brightness) stacks.
The saturation stacks were used to set the boundaries of the
nuclear chromatin intensity and thresholded by red binary
color to include all chromatins in epidermal cells. The level of
chromatin condensation and area percentages were represented
by the level of white color that occurred within the nucleus.
Reticular Dermis Integrated Density and
Area Percentages
Quantifying intensity of staining and area percentages of
reticular dermis sections (different PMIs), routinely stained by
HE, was done by transforming the image into gray image and
then masking the positive areas by red binary color.
The average area percentages of reticular dermis fibers were
selected and masked by red binary color. Then, the area of the red
binary color was measured and expressed as average area in
relation to the area of the standard measuring frame and expressed
by area %.17
Sebaceous Gland Nuclear Areas and Their
Chromatin Condensation
Areas and intensity of nuclear chromatin of sebaceous gland
nuclei were measured individually for each nucleus in routinely
stained slides. The average areas and nuclear chromatin intensities
of nuclei were selected and masked by red binary color.18
Statistical Analysis
Data are expressed as mean SD. The significance between
PMI groups was determined using analysis of variance followed
by post hoc test (Dunnett test) for statistical analysis of all possible
pairwise comparisons (with 0 hours PMI). P < 0.05 considered to
be significant.
RESULTS
Progressive morphological changes of the skin, its append-
ages, and underlying muscles in relation to PMI were classified
according to the histological structure of the skin, as follows:

FIGURE 1. ah, Epidermal cell layers showing different levels of histopathologic nuclear chromatin condensation changes at different PMIs.
AH, Corresponding s aturation stacks used for measuring nuclear chromatin intensities and area percentages. Area percentages of
epidermal nuclear chromatin were 1.99%, 2.37%, 6.07%, 8.72%, 8.22%, 9.86%, 5.61%, and 5.21%. Figure 1 can be viewed online in color
at www.amjforensicmedicine.com.

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Am J Forensic Med Pathol Volume 00, Number 00, Month 2017 Forensic Image Analyses of Skin and Muscles

Epidermis
At time of death, no morphological changes could be
detected in the epidermis (Fig. 1a). The stratum spinous and
basalis appeared distended and ballooned, as well as the presence
of condensation of nuclear chromatin at different PMIs (Fig. 1, bh).
Data revealed from the examination of saturation stacks of examined
fields, for determining the morphological changes in the epider-
mal nuclei, are briefly illustrated in Figure 1, AH and Table 1.
At 0 hours PMI, uncondensed nuclear chromatin could be
detected in most of nuclei, with vesicular nucleoli appearance.
At 8 to 16 hours PMI, there was appearance of nuclear chromatin
condensation, in some nuclei showing a continuous ring conden-
sation of nuclear chromatin, mainly at the interior aspect of the
nuclear envelope. At 24 to 40 hours PMI, elevated nuclear chro-
matin condensation reached the peak at 40 hours PMI. At 48 to
72 hours PMI, significant decreases in the nuclear chromatin
condensation were found (Fig. 2A).
Statistically, 1-way analysis of variance revealed a significant
difference (P < 0.001) in the nuclear chromatin intensities at
different PMIs. In specification, compared with 0 hours PMI,
the use of post hoc test (Dunnett test) revealed no significant
difference between 0 hours PMI and either 8 or 16 hours PMI
(P = 0.961 and 0.063, respectively). In contrast, a high significant
difference could be detected with 0 hours and 32 and 40 hours
PMI (P < 0.001), whereas P = 0.004 at 72 hours PMI. Meanwhile,
statistical analysis of epidermal nuclear chromatin area percent-
ages showed no significant difference between 0 and 8 hours
(P = 0.509). More or less significant difference between 0 hours
and 16 and 24 hours (P = 0.032 and 0.007, respectively) was
found. A high significant level of difference could be detected at
32 and 40 hours PMI (P < 0.001).
Dermis
Reticular Dermis
At 0 hours, a normal histological structure of the reticular
dermis consisted of dense irregular connective tissue containing
collagen and elastin was described (Fig. 3a). At 8 to 16 hours
PMI, there were main histopathologic alterations represented by
a focal degeneration of the reticular dermis (Fig. 3, b and c). With
increasing PMI, the degeneration became diffuse (Fig. 3, d and g).
By the end of 72 hours, the dermis began as coagulated masses
of reticular dermis, which in some areas appeared disintegrated,
associating with nuclear losses in most cells (Fig. 3h). Quantitative
analysis measurement of reticular dermis (dermal collagens)
area percentages was illustrated (Fig. 3, A and H; Table 1).
Statistically, no significant difference was found between 8 and
16 hours PMI (in relation to 0 hours). However, a high
significant difference could be detected at 24 to 72 PMI and
0 hours PMI (P < 0.001) (Fig. 2B).
Sebaceous Glands
At the time of death, no obvious morphological changes
could be detected in the sebaceous gland cells. The latter appeared
foamy with almost intact nuclei, mainly until 16 hours PMI
(Fig. 4, a and c). At 24 to 40 hours PMI, the foamy cytoplasm is
gradually disintegrated with severe nuclear changes of
sebaceous cells (Fig. 4, d and f ). At 48 to 72 hours PMI,
sebaceous gland cells disintegrated, and many of the nuclei were
totally lost (Fig. 4, g and h). Morphometrically, the average
areas (in m2) and condensation of nuclear chromatin (pixels) of
sebaceous cells are illustrated in Table 1 and Figure 2C. At
0 hours PMI, the nuclear areas were approximately 35 m2, and
the nuclear chromatin condensation was 9000 pixels (Fig. 4A).
The areas and intensities of nuclear chromatin 72 hours PMI
(Fig. 4H) were severely reduced to reach less than 10 m2 and
less than 2000 pixels, respectively. Significant differences in
sebaceous gland nuclei areas among different-hours PMI were
found (P < 0.001) (Figs. 2C and 4B, G).
Hair Follicles
The hair follicle appeared normal at 0 to 16 hours PMI
(Fig. 5A) except for minimal separation between inner and outer
follicular wall at 8 hours PMI (Fig. 5B). At 24 hours PMI,
minimal degenerative change appeared on the medulla and
cortex of hair shaft, and moderate changes could be detected in
the follicular wall (Fig. 5C). At 32 to 72 hours PMI, a separation
between cuticle and follicular wall, severe degenerative changes
and necrosis in the follicular wall with more or less degeneration
of cortex, medulla, and hair shaft were noted (Fig. 5D).

TABLE 1. Morphometric Comparison of the Epidermal Nuclear Chromatin Condensation and Area Percentages, Reticular Dermis
Area Percentages, and Sebaceous Glands Nuclear Chromatin Condensation and Areas at Different PMIs (Mean SD)

Epidermis Dermis
Epidermal Cell Epidermal Cells Sebaceous Cell Areas of Individual
Nuclear Chromatin Nuclear Chromatin Reticular Dermis Nuclear Chromatin Sebaceous Cell,
PMI Area, % Condensation/Pixels Area, % Condensation/Pixels Nuclei m2
0h 3.67 2.20 5,370,759.1 3,651,921.1 46.50 4.91 8764.56 2513.05 34.37 9.86
8h 4.83 4.09 6,160,603.9 4,718,820.1 48.72 4.87 7170.62 2540.00* 28.12 9.96*
16 h 5.83 2.29 8,024,649.6 3,151,110.5 47.87 4.21 5173.01 2535.55* 20.15 10.05*
24 h 6.20 4.04 8,542,979.6 5,564,963.3 52.98 5.93* 4313.85 2710.20* 16.92 10.63*
32 h 7.44 4.02* 10,250,533 5,541,783.4* 52.99 6.70* 3064.85 2115.89* 12.019 8.30*
40 h 7.75 5.12* 10,671,580 7,043,529.54* 52.47 4.59* 2477.84 2073* 9.42 8.18*
48 h 6.24 2.31 8,591,223.21 3,181,071.41 54.97 9.08* 2483.79 1535.72* 9.10 6.31*
72 h 6.63 3.07 9,133,720.95 4,250,479.32 58.56 8.04* 1986.03 1944.628792* 7.32 7.62*
Epidermal nuclear chromatin condensation was measured in specified areas (1  6 inches).
*Significant at P < 0.001 (compared with those at 0 hours PMI).
Significant at P < 0.01 (compared with those at 0 hours PMI).
Significant at P < 0.05 (compared with those at 0 hours PMI).

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El-Nahass et al Am J Forensic Med Pathol Volume 00, Number 00, Month 2017

FIGURE 2. A, Epidermal nuclear chromatin intensities at different PMIs with the highest intensity at 40 hours PMI. Analysis of variance,
P < 0.05, 0.001, compared with 0 hours. B, Reticular dermis area percentages at different PMIs showing a high significant difference
between 0 hours and 27 to 72 hours PMI (P < 0.001). C, Sebaceous nuclei areas had a dramatic decrease relative to different PMI hours
(P < 0.001).

Hypodermis, Nerve Endings, Blood Vessels, and
Underlying Muscles
Examination of the hypodermis showed lobules of adipose
tissue were surrounded by loose connective tissue with absence
of pathological changes at 0 to 16 hours PMI (Fig. 5E). At 24
to 32 hours, there was a mild decrease in the thickness of
hypodermis with preserved outline of adipose tissues as well as
pyknotic nuclear changes (Fig. 5F). At 40 to 48 hours PMI, there
was a severe decrease in hypodermis thickness in association with
shrinkage of adipose tissues at different PMIs (Fig. 5G). At
72 hours PMI, destruction of adipose tissues with loss of their
boundaries was detectable (Fig. 5H).
Examination of red blood cells showed intact, normal-shaped
red blood cells with presence of central pallor in most cells at
0 hours PMI (Fig. 5I). At 8 hours PMI, intact red blood cells
with slight dysmorphic changes could be detected (Fig. 5J). At
16 to 32 hours PMI, a lot of cells changed from discoid to

FIGURE 3. ah, The reticular dermis showing variable degrees of reticular dermis degenerative changes and coagulation. BH, Area
percentages of reticular dermis of binary images corresponding to (ah) different PMI hours. Area percentages of reticular dermis are
43.32%, 48.49%, 48.97%, 51.52%, 52.16%, 52.79%, 58.40%, and 65.59%. Figure 3 can be viewed online in color at
www.amjforensicmedicine.com.

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Am J Forensic Med Pathol Volume 00, Number 00, Month 2017 Forensic Image Analyses of Skin and Muscles

FIGURE 4. ah, Sebaceous gland cells showing different levels of nuclear changes (areas and intensities) at different PMI hours. AH, Binary
images corresponding to ah used for individual measurement of sebaceous cells nuclei areas (in m2) and intensities of their nuclear
chromatin. The averages of sebaceous nuclei areas are 39.07, 20.33, 18.31, 17.35, 16.871, 9.31, 9.28, and 7.5 m2. Figure 4 can be viewed
online in color at www.amjforensicmedicine.com.

elliptical shape; mixture of intact and lysed red blood cells could
be found with absence of central pallor (Fig. 5K). At 40 to
72 hours PMI, several red blood cells were completely lysed and
unrecognizable (Fig. 5L).
Skeletal muscle layer beneath the hypodermis layer was
observed. A well-defined striation of skeletal muscles could be
easily detected with more or less normal peripheral nuclei at 0 to
8 hours PMI (Fig. 5M). The divinity of those striations is
gradually decreased from 16 to 24 hours PMI (Fig. 5N). At 32 to
48 hours PMI, ill-defined striation of skeletal muscle could
be seen in association with variable degrees of coagulation of
skeletal muscle cytoplasm and pyknosis of peripheral nuclei
(Fig. 5O). At 72 hours PMI, it was difficult to detect cross
striation of skeletal muscles in most areas with disintegration
of their cytoplasm and peripheral nuclei (Fig. 5P).
The peripheral nerve endings either in the dermis or the
underlying musculature exhibited normal histological structure
including myelinated axons (Fig. 5Q) at 0 to 8 hours PMI.
Thereafter, degenerative changes, vacuolation, and demyelination
could be detected from 16 to 32 hours PMI (Fig. 5R). Progressive
changes and necrosis could be found until 48 hours PMI
(Fig. 5S). At 72 hours PMI, complete demyelination of peripheral
nerve endings was easily detected (Fig. 5T).
DISCUSSION
Histopathologic examination provides a comprehensive view
not only of disease diagnosis but also in forensic practice. Exam-
ination of routinely stained HE slides preserves the underlying
tissue architecture, providing an important evaluation in death
investigation,19 placental ischemic changes,20 and breast cancer
metastasis examination. In the latter, the accuracy for diagnosis
of sentinel lymph node metastasis associated with breast cancer
was 97.0% by examination of stained paraffin section examina-
tion.21 Costs of routine histopathologic examination were very
low compared with other techniques. Such finding was consistent
with that of Lochmuller and Pestaner,19 who stated that routine
histopathologic examination was reliable with a low cost.
Forensic histopathology is an important tool in forensic
medicine because it illustrates the microscopic analysis associated
with cellular changes throwing light on the cause of death.22 In the
current study, we tried to review the morphometric and histopath-
ologic analyses in the skin and its underlying muscles. Those
alterations were investigated at different hours after death. In
such circumstance, the use of ImageJ software is considered a
valuable method for evaluating histopathologic examination
of the skin and its underlying tissues.18 Many environmental factors

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El-Nahass et al Am J Forensic Med Pathol Volume 00, Number 00, Month 2017

FIGURE 5. Postmortem interval histopathologic alterations in skin layers relative to the time of necropsy. Hair follicles showed (A) a normal
histological structure at 0 hours PMI, (B) a mild separation between inner and outer follicular walls at 8 hours PMI, (C) moderate
degenerative changes in the follicular wall at 24 hours PMI, and (D) a separation between hair cuticle and follicular wall at 32 hours PMI.
Adipocytes showed (E) normal histological structure at 8 hours PMI, (F) a slight corrugation in adipocyte wall at 16 hours PMI, (G) severe
degenerative changes of the cytoplasmic wall and nuclear pyknosis at 32 hours PMI, and (H) a complete destruction of the cytoplasmic wall
at 48 hours PMI. Red blood cells showed (I) normal morphological structure at 0 hours PMI, (J) dysmorphic changes in most cells at 8 hours
PMI, (K) elliptical cells at 16 hours PMI, and (L) a complete lysis of most cells at 40 hours PMI. Skeletal muscles showed (M) normal striation at
8 hours PMI, (N) weak striation, (O) coagulation and nuclear pyknosis, and (P) a complete disintegration of cytoplasm and nuclei. Peripheral
nerve endings showed (Q) normal structure at 0 hours PMI, (R) a slight separation between neurofibrils at 16 hours PMI, (S) demyelination and
nuclear pyknosis at 32 hours PMI, and (T) a complete demyelination of nerve fibers at 48 hours PMI (HE stain, original magnification
1000). Figure 5 can be viewed online in color at www.amjforensicmedicine.com.

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Am J Forensic Med Pathol Volume 00, Number 00, Month 2017
including temperature, humidity, and insect activity play important
roles in detection of cutaneous autolysis in many studies.23,24
Comparing epidermal nuclear area percentages and intensi-
ties of skin, it was found that there was an increase in parameters
as a result of nuclear chromatin condensation, which started from
8 hours until 40 hours PMI. At 48 and 72 hours PMI, parameters
began to decrease as a result of degeneration and disintegration of
nuclear chromatin. Previous literature showed that 3 stages of
nuclear chromatin condensation in cell-free apoptotic system
could be detected and were preceded by uncondensed chromatin
stage followed by a ring condensation stage, necklace condensation
stage, and nuclear collapse or disassembly stage.25 Previous in vivo
studies stated that the mechanism explaining the rapid stage 1 ring
condensation was clearly unknown. Different unidentified factors,
essentially some enzymes, are responsible for modifying chromatin
or caspase formation.26 Furthermore, previous biochemical frac-
tionation experiments showed that fraction 1 was responsible for
the ring condensation stage.27
Morphometric analysis of dermal collagen area percentages
revealed a noticeable increase relative to progressed PMI that
was attributed to the formation of coagulated masses of fibers,
which was clearly observed at 72 hours PMI. Histopathologically,
degeneration in the dermal collagen started at 8 hours PMI, and
with increasing PMI, the degeneration becomes obvious and gen-
eralized. In agreement, previous studies reported degeneration in
the dermis at 6 to 9 hours after death, but the disintegration of
the dermis occurred 18 hours after death.6 The importance of
morphometric analysis of collagen and reticular fibers had
been studied by several authors.16,28,29 Rich and Whittaker30
found that hue analysis of collagen fibers provides a powerful
tool for collagen fiber structural analysis.
Another important histological finding in skin sample ex-
amination was that of sebaceous glands. Histopathologic and
morphometric results of sebaceous glands appeared approxi-
mately 24 hours PMI, and degeneration increased with increased
hours of PMI. The degeneration was severely detected in sebaceous
glands than other skin structures, including the hair follicles. This
is attributed to its excretory function, which is facilitated by
their deterioration.6,24
As a result of the present work, examination of hair follicles
showed relatively persistent state of histopathologic alterations
compared with other skin structures especially sebaceous glands.
The degenerative changes appeared 24 hours PMI with moderate
degrees. At 32 hours PMI, the main pathological alterations
appeared in the follicular wall with moderate changes in the hair
shaft. Similar results were found by Bardale et al,6 who found that
hair follicles appeared normal until 18 hours after death.
In the current study, examination of the dermis, hypodermis,
and underlying skeletal muscles including nerve endings revealed
valuable pathological alterations in relation to PMI. Limited
literature could be found, especially those that illustrate adipocytes
and nerve ending degradation. In the present work, the cytoplasmic
membrane of adipose cells and wall nerve endings were intact until
16 hours PMI, and complete cytoplasmic and nuclear destruction
of adipocytes and severe vacuolation of nerve endings were
detected at 48 hours PMI.
The main microscopic alterations of red blood cells inside
blood vessels coursing either through the dermis, hypodermis,
dermal-hypodermal boundary, or underlying muscular layer are
based on examination of their shape, integrity, and presence of
central pallor. Clear degenerative changes of red blood cells could
be easily detected at 16 hours PMI until complete lysis of a lot of
cells at 40 hours PMI and increased until cell lysis at 72 hours
PMI. Previous literature showed that red blood cells are still
identifiable until 18 hours postmortem, and they completely lysed
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Forensic Image Analyses of Skin and Muscles
at 24 to 48 hours PMI.23,31 Contritely, different types of cells, eosin-
ophils and monocytes, were identifiable at much longer time and
reached up to 72 hours. Neutrophils were found at up to 96 hours,
whereas lymphocytes were considered the most stable cells as they
were present up to 120 hours in nonrefrigerated cases.32
The present study clearly demonstrated that histopathologic
alternations in skins underlying muscles may be considered a tool
for PMI determination, which is coming in parallel with other
studies that found that muscle disintegration could be detected
by using biochemical techniques including Western blotting or
zymography showing variable protein band appearance in relation
to postmortem time.33 In the present study, the striation of those
muscles could be identified until 8 to 16 hours PMI, whereas the
initial degradation processes of skeletal muscles started at 16 hours
PMI. Changes increased upon accumulated degree-hours after
death. Previous reports identified particular proteins including
desmin, cTnT, and calpain 1 and their degradation products,
which are used as markers in determining the time intervals and
postmortem decomposition after death.29 Similarly, Liu et al34,35
found that actin filament disintegration began at 24 hours PMI
and extended with a gradual decrease associated with prolonged PMI.
Histopathologic changes were more or less similar to previ-
ous studies that described postmortem alterations of the skin;
however, the present results appeared later than the other studies,
which is attributed to the changes in the environmental difference
at which the samples are collected (average temperature 15C).
The extreme environmental factors influenced the speed of
decomposition. In hot weather, rapid cutaneous decomposition
occurs. In contrast, cold weather preserves the skin integrity.24,36
The current study was conducted in restricted or limited
environmental condition as previously described; therefore, ad-
ditional specialized studies will be needed to evaluate and compare
changes in different environmental conditions with special refer-
ences to nerve endings and adipose tissue changes not only in the
skin but also in other sites. In conclusion, this study might be used
to evaluate early and late time of death by determining PMI using
morphometric parameters in skin and its underlying muscles.
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