Indian Journal of Pure & Applied Physics
Vol. 42, March 2004, pp 189-194
Surface morphology of gall bladder stones
V R Singh & Palwinder Kaur
Instrumentation and Sensors Group, National Physical Laboratory, New Delhi 110 012
Received 22 July 2003: accepted 13 November 2003
Gall bladder stones are crystalline structures formed by concretion or accretion of normal or abnormal bile
constitutents. These stones are normally made of cholesterol, calcium, phosphate, carbonate and proteins. The aetiology of
gall bladder stones is multifactoral. Scanning electron microsscopy (SEM) is used here to investigate the surface features of
the mechanically fractured part of the stone samples. Micrographs exhibit a non uniform polycrystalline surface without
specific grain boundaries with porosity, leading to loose bounding of crystals and hence low specific gravity. These results
pertaining to surface characterisation are very useful for understanding the disintegration process of stones inside or outside
the human body.
[Keywords: Gall bladder stones, Morphology, Surface morphology, Disintegration process]
IPC Code: G 01N 13/10
1 Introduction
The organs of the digestive system perform a vital
process. These organs prepare food for absorption and
for use by the millions of the body cells. Gall bladder
is the part of the digestive system, which stores the
bile that enters it by way of the hepatic and cystic
ducts, and during digestion, it ejects the concentrated
bile into the duodenum1. Lithiases (the stone
formation) is a major disease associated with the
human gall bladder. When cholesterol concentration
exceeds the solubility limit in the lipid bilayers of
miscelles, deposition of cholesterol takes place. This
leads to nucleation of crystalline cholesterol. The
causative factors for the occurrence of gallbladder
stones are hereditary, infection and diet. These stones
are frequently twined, cracked, yellowish white or
yellowish brown in colour and have fine granular
crystalline or non-crystalline-structure2,3. These stones
on the basis of their chemical composition are
classified as: pure cholesterol, pure calcium and
combination. Dielectric properties in gall bladder
stones are found due to the presence of silicon
dioxide, while the variation in ultrasonic properties is
because of their complex structures4-14.
The electron microscopy has become an
indispensable tool for research and development and
has a key role to play in the field of environmental
and forensic science, industrial development, genetic
engineering, medical and biological fields to examine
cells, tissues and micro-organisms. Materials science
studies on lattice defect, inclusions, precipitates,
phase transitions and other technical fields associated
with semiconductors also involve the use of electron
microscopy. SEM is also commonly used in hospitals
and medical centres for clinical diagnostic purposes.
This system gives a three-dimensional quality to its
pictures, even although its limited resolution is around
100 Å. The whole of the specimen is not illuminated
at the same time and the electron beam is focussed to
needle sharpness so that it strikes an area of only
about 10 nm radius at any instant. Several million
electrons strike the given sample in one second and
scan the sample.
In the present work, surface characteristics of
different gall bladder stones have been studied with
scanning electron microscope system. This study gives
very useful information of the surface structure of gall
bladder stones. This is found to be helpful in
understanding the physical mechanisms of the
disruption of different stones inside or outside the body.
2 Materials and Method
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INDIAN J PURE & APPL PHYS, VOL 42, MARCH 2004
In the present study, twenty biological (gall
bladder) stones were collected from different hospitals
in New Delhi such as Lok Nayak Jai Prakash Narain
Hospital, All India Institute of Medical Sciences, and
Bara Hindu Rao Hospital. These stones were obtained
through surgical operations on different patients. Four
stone samples (Pg1, Pg2, Pg3 and Pg4) were considered
for investigating the physical properties such as the
dimension, shape, finish, porosity, specific gravity,
and micro and macro structures with SEM technique.
The scanning electron microscope gives useful
information about the structure of the specimen at
high resolution and has good focus over a wide range
of specimen surfaces. The system used produces clear
images of the specimens ranging from objects visible
with the naked eye to the structures as small as size of
several angstroms. Fig. 1 shows the photographs of
different gall bladder stones used for SEM analysis.
The samples were fixed on the small circular
discs separately. These stone samples were coated
with gold layer (100-200 Å) to avoid any charging
effect. SEM coating unit (make Poland Equipment
Ltd. E 5000) was used here. SEM, JEOL model JSM840, was used to characterise the surface structure of
samples. In the experiment, electron beam emitted
from the electron gun was focussed on to the surface
of the specimen by an electron lens. The number of
electrons striking on the unit area of the specimen was
determined by the diameter of the electron probe. Fig.
2 shows the schematic diagram of the system used.
The probe was moved horizontally by supplying
current to the horizontal-scanning coil located in the
electron beam path. The secondary electrons emitted
Fig. 1 ⎯ Photographs of GB stones used in the present
investigation
Fig. 2 ⎯ Layout of SEM set-up
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SINGH & KAUR: SURFACE MORPHOLOGY OF GALL BLADDER STONES
Table 1 ⎯ Physical properties of gall bladder stones
Sample No.
Form
Colour
Lustre
Steak
Hardness on
mho’s scale
Observed specific
gravity
Pg1
Amorphous
Yellowish
Resinous/Silky
Yellow
3.0
Low
Pg2
Amorphous
(Nodular)
Brownish black
Earthy
Yellow
2.5
Low
Pg3
Crystalline
Brownish white
Earthy
White
2.0
Low
Pg4
Amorphous
Yellowish Brown
Earthy
Yellow
1.5
Low
Table 2 ⎯ SEM data for gallbladder stone samples (Pg1, Pg2, Pg3
and Pg4) at an accelerating voltage of 10 kV
Sample
Magnification
Particle size
(Range) (μm)
Working distance
(WD) (mm)
Pg1
×12 .
1000
34
×2000
10
34
×5000
1
34
×16 .
1000
32
×2000
10
32
×5000
1
32
×12 .
1000
31
×1000
10
32
×2500
10
31
×12 .
1000
31
×2000
10
31
×5000
1
31
Pg2
Pg3
Pg4
from various positions of the specimen surface when
scanned by the probe were determined by the
secondary electron detector and the signal induced in
the detector was sent to the video amplifier, where it
was amplified and level controlled to obtain an
optimum contrast and brightness image on the CRT
(Cathode Ray Tube). The video output signal was
then supplied to the control grid of the cathode ray
tube, which was synchronised with probe scanning.
The signal modulates the raster thereby displaying a
scanning image on the screen.
3 Results and Discussions
Table 1 presents the physical observations of
different gall bladder stone samples. These stones are
found to be earthy (dull) lustre but differing in
colouring in colour, steak and hardness. All the
samples are soft because of their low specific gravity
and hardness. Their hardness on mho’s scale is around
2.
Fig. 3 ⎯ Micrographs of GB sample Pg1 at different
magnifications
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INDIAN J PURE & APPL PHYS, VOL 42, MARCH 2004
SEM experiments were performed at an
accelerating voltage of 10 kV and probe current of
10-11 Ampre and resolution of the order of 60 Å. The
micrographs depict the deposition of crystals, which
form the irregular and unoriented stone. Table 2
shows the observations of gall bladder stones with
SEM at different magnifications with working
distance (focussing distance) and depth of field.
Micrographs of different magnifications give the
surface features of the samples used, which are
helpful in breaking these stones. The surface
morphology of specimen Pg1 (gall bladder stone) is
given in Fig. 3(a,b,c). Its surface appears slightly
smooth and all crystals are more or less of the same
shape but size of crystals is large. There are cavities
on its surface, which result in a non uniform
discontinuous rough surface as shown at
magnification×5000. Fig. 4(a,b,c) shows the surface
structure of the sample Pg2. It possesses
polycrystalline uneven surface and at magnifica-
Fig. 5 ⎯ Micrographs of Pg3 sample of GB at different
magnifications
Fig. 4 ⎯ SEM pictures of internal structure of GB sample Pg2 at
different intensities
tion×2000, a number of pits were observed. Crystals
inside the body are without any grain boundaries,
SINGH & KAUR: SURFACE MORPHOLOGY OF GALL BLADDER STONES
which causes heterogeneous nature. At magnification
×5000, some crystals appear in stick type growth
while others have no definite shape. The
microstructure of sample Pg3 is given in Fig. 5(a,b,c).
Its surface appears flat with black spots on it. At
magnification ×1000, there are cracks on the surface,
which exhibits an amorphous type of pattern with
193
many islands. These islands are formed by small
polycrystalline grains. Fig. 6 (a,b,c) shows the surface
morphology of stone sample Pg4. It gives better
surface texture as observed at magnification ×12. All
crystals are of different sizes and shapes with cavities
without any definite pattern because deposition of
these stones also depends upon the diet intake of the
patient. These results of the physical properties and
SEM analysis confirm that the gall bladder stones can
be broken inside or outside the human body.
4 Conclusions
Surface structure study of different gall bladder
stones proves that gall bladder stones are soft and
smooth with large cavities on them. Different crystals
normally are of different sizes and shapes and cavities
on them form loose bonding which helps crushing
these stones. The present work would be helpful in
understanding better the mechanisms involved in the
disintegration of gall bladder stones, after correlating
the micrographs with the nature of particular stone
samples.
Acknowledgement
We are thankful to Dr. Kusum Gupta, Bara Hindu
Rao Hospital, Delhi, for fruitful suggestions in the
paper and for the help in the procurement of the stone
samples used in the study.
References
Fig. 6 ⎯ SEM analysis of Pg4 sample of GB at magnifications of
×12, ×2000 and ×5000
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