CROMATINA SEXUAL

1. J Indian Acad Forensic Med. April-June 2011, Vol. 33, No. 2 ISSN 0971-0973
143
Original Research Paper
Use of Hair Root Sheath for Barr body Determination
*Harpreet Singh, ** O.P. Aggarwal, *** Arsalaan F. Rashid
Abstract
Determination of sex is useful in Forensic Medicine. Barr bodies can be seen hair root sheath
cells. The presence of barr bodies indicates that sex of the person is female. They are inactive X
chromosome. The Barr bodies are also known as sex chromatin. Hair can present in the crime scene.
Hair is trace evidence. The stain used was aceto-orcein. The percentage of barr bodies was found to be
28-49 %. The study was carried out for eight months and it was found that barr bodies persisted for eight
months. It can be present anywhere at the crime scene. Hair of the accused can be present in hands of
victim due to cadaveric spasm. Hair of victim can be found on the clothes of the accused. There is
average fall in percentage of barr bodies with the passage of time due to the start of decomposition
changes in the root sheath cells.
Key Words: Root Sheath, Barr body, Hair, Trace evidence, Aceto-Orcein Stain, Sex Chromatin
Introduction:
The sex can be determined from the hair by
the microscopic examination of the sex
chromatin. Sex chromatin was first described in
1947 by Barr and Bertam. [1] The sex chromatin
is of two types i.e. X-chromatin and Y-chromatin.
X-chromatin is also known as Barr body. Y-
chromatin is also known as Y chromosome and
Y body. The aims and objectives of the present
study were described as:
1) To determine the female sex by
demonstration of Barr bodies in hair root
sheath cells of the normal females.
2) To determine the time interval to which
the sex could be identified from hair
after plucking from scalp.
Methods:
The present study conducted on the
scalp hairs obtained from the cadavers coming
to the mortuary of M.M.I.M.S.R Mullana. There
were 50 females. Hair root sheath cells studied
for Barr bodies. In the present study we plucked
nine hair from the scalp of each dead body. After
plucking the hair they were stored in the plastic
bags. The plastic bag was labelled. At the same
time the Performa of case was filled.
Corresponding Author:
*Assistant Professor
Deptt. of Forensic Medicine, MMIMSR
Mullana Ambala
E-mail: afrashid@gmail.com
** Prof. & Head
***Resident
Deptt. of Forensic Medicine, MMIMSR
Mullana Ambala
We used the Aceto-orcein stain for the
staining of Barr bodies.
Demonstration of Barr body by Aceto-orcein
staining: (Sanderson and Stewart 1961) [2]
Special Reagents Required: Stock aceto-
orcein solution: It was prepared by dissolving
the synthetic orcein (1gm) and Glacial acetic
acid (45ml). This solution was boiled and
allowed to get cool. At the end it was filtered by
using the filter paper.
2. Staining solution: Staining solution was
prepared from the stock aceto-orcein solution.
55 parts of the distilled water was added to 45
parts of the stock aceto-orcein solution. The
staining solution was filtered at time of staining
of the slides. Method of preparation of the slide
of the root sheath cells of the hair.
(1) Hair was taken out from the plastic bag by
using the forceps.
(2) The hair was kept on the slide. With the help
of blade the bulb of the hair was cut. The
root sheath slipped of the shaft.
(3) Without fixing, a drop of stain was placed on
the root sheath and then No.1 cover slip laid
on it.
(4) A layer of filter paper was over the cover slip
and pressed down by drawing the thumb
across from one end to another.
(5) Finally, the slide was examined with x40 or
x100oil immersion objective lens on the
Olympus microscope. Hundred clearly
visible nuclei were examined. Only those
nuclei were counted which were free from
the indentations and which did not show any
overlapping.

2. J Indian Acad Forensic Med. April
The Barr bodies in the marginal position
were counted. The oil used was cedar wood oil.
Results were entered in the Performa.
Observation:
Table 1: Age Distribution of Females
Age groups No of cases Percentage
Up to 10 years 3
11-20 years 12
21-30 16
31-40 11
41-50 5
51-60 3
61-70 0
Total 50 100%
Range 2 days to 60 years
Mean 29.18
±SD ±12.83
Fig: 1 Mean of Barr Bodies of Females
Table 2: Percentage of Barr Bodies at different Time Intervals among Females:
(%) 1
st
day
after
1 mth
after 2
mths
41-50 33 29 5
31-40 15 26 31
21-30 2 4 11
11-20 - - 2
1-10 - - -
0 - 1 1
Total 50 50 50
0
10
20
30
40
50
day
1
2 4 6 8
M e a n
M o n t h l y I n t e r v a l s
J Indian Acad Forensic Med. April-June 2011, Vol. 33, No. 2
144
The Barr bodies in the marginal position
were counted. The oil used was cedar wood oil.
Results were entered in the Performa.
f Females:
Percentage
6%
24%
32%
22%
10%
6%
0
100%
2 days to 60 years
f Females:
Discussion:
The percentage of Barr bodies in the
female root sheath cells were 28
42.2%). Culbertson et al [3] (1969) found 65.1%
Katz and Wright [4] (1970) found the 60
Das et al [5] (2004) found 42
are of present study is as 28%
Barr bodies on day one, 0%-46% (mean 38.42)
after one month, 0%-44% (mean 34.70) after
two months, 0%-41% (mean 30.08) after three
months, 0%-40% (mean 25.30) aft
months, 0%-39% (mean 20.90) after five
months, 0%-35% (mean 17.34) after six months,
0%-30% (mean 12.82) after seven months and
0%-27% (mean 7.96) after eight
shown in the Table II. After eight months, sex
could be determined in 10 female
Torr (1956) [6] observed for the longest period
(23 days). Nagamori and Takeda
this for 32 weeks in their study.
References:
1. Barr ML and Bertram EG. A morphological distinction between the
neurons of male and female and behavior of the nucleolar satellite
during the accelerated nucleoprotein synthesis. Nature 1949;
163:676-677.
2. Sanderson AR and Stewart JSS. Nuclear sexing with Aceto
orcein. Brit Med J 1961; 2:1065.
3. Culbertson JC, Breslau NA, Moore MK and Engel E.
chromatin determination from hair. JAMA 1969 Jan 20; 207(3):560
561.
4. Katz MM and Wright SW. The use of hair root sheath for X
chromatin determination. The Journal of Pediatrics 1970; 76(
295.
5. Das N, Gorea RK, Gargi J and Singh JR.
pulpal tissue. JIAFM 2004; 26(2):50-54.
6. Dixon AD and Torr JBD. Post–mortem persistence of sex
chromatin. J Forens Med 1956; 3:161.
7. Nagamori H and Takeda K. Sex determination from plu
human hair without epithelial root sheath .III. Fluorescent Feulgen
reaction using Acriflavine. Forensic science international 1981;
17:85-90.
Table 2: Percentage of Barr Bodies at different Time Intervals among Females:
No. of cases
after 3
mths
after 4
mths
after 5
mths
after 6
mths
after 7
mth
2 - - - -
29 20 7 4 -
9 17 26 24 18
6 3 3 5 9
1 1 1 1 4
3 9 13 16 19
50 50 50 50 50
ISSN 0971-0973
The percentage of Barr bodies in the
female root sheath cells were 28-49% (mean
(1969) found 65.1%.
(1970) found the 60-90%.
Das et al [5] (2004) found 42-52%. The results
are of present study is as 28%-49% (mean 42.2)
46% (mean 38.42)
44% (mean 34.70) after
41% (mean 30.08) after three
40% (mean 25.30) after four
39% (mean 20.90) after five
35% (mean 17.34) after six months,
30% (mean 12.82) after seven months and
after eight months as
After eight months, sex
could be determined in 10 females. Dixon and
Torr (1956) [6] observed for the longest period
Nagamori and Takeda (1981) [7] find
weeks in their study.
A morphological distinction between the
neurons of male and female and behavior of the nucleolar satellite
during the accelerated nucleoprotein synthesis. Nature 1949;
Nuclear sexing with Aceto-
Culbertson JC, Breslau NA, Moore MK and Engel E. Sex
chromatin determination from hair. JAMA 1969 Jan 20; 207(3):560-
The use of hair root sheath for X–
chromatin determination. The Journal of Pediatrics 1970; 76(2):292-
Das N, Gorea RK, Gargi J and Singh JR. Sex determination from
mortem persistence of sex
Sex determination from plucked
human hair without epithelial root sheath .III. Fluorescent Feulgen
reaction using Acriflavine. Forensic science international 1981;
after 7
mths
after 8
mths
-
-
18 10
6
6
19 28
50 50