Morphology Based Detection of Abnormal Red Blood Cells in Peripheral

Blood Smear Images

S. Kulasekaran1, Feminna Sheeba2, Joy John Mammen3, B. Saivigneshu2, and S. Mohankumar2
1
Healthcare Technology Innovation Centre, Chennai, India
2
Department of Computer Science, Madras Christian College, Chennai, India
3
Department of Transfusion Medicine & Immunohaematology, Christian Medical College, Vellore, India

Abstract— Red blood cells are the most abundant type of
blood cells in the human body, delivering oxygen to body tis-
sues.The count of these vital cells is often the first step done in
analyzing a patient’s pathological condition. Normal RBC’s
are biconcave in shape with a central pale area, and any devia-
tion in size, shape, volume, structure or color represents an
abnormal cell. Such abnormalities are detected by viewing the
blood-smear images through a microscope, a time consuming
and error-prone method. This process can be automated by
analyzing the individual cells in a peripheral blood smear
image and segmenting the cells using appropriate segmenta-
tion techniques. The proposed study aims at Morphology-
based detection of abnormal red blood cells in peripheral
blood smear images, based on their size and shapes. Abnor-
malities such as Anisocytosis, Macrocytosis and Microcytosis
are detected based on the size of the RBCs. Variations in the
shape of RBCs couldindicate various abnormalities. Convex
hull based detection of speculated RBCs, is carried out in
Acanthrocytosis and Echinocytosis. The condition Eliptocyto-
sis, where some of the RBCs turn elliptical is detected using
Houghman Transform. In the abnormality called Rouleaux the
RBCs appear as stack of coins, which are detected by applying
a watershed algorithm to individual stacks and counting the
number of cells in the stack. Sickle cell anemia is another
common condition in people, where few RBCs are sickle or
crescent shaped and this shape is determined using the round-
ness factor. Codocytes resemble a bull’s eye, and can be identi-
fied by examining if the segmented RBCs have rounded areas
within the cell. Dacrocytes are tear drop RBCs, which can be
detected by analyzing the extreme points of the cell. The expe-
riment was conducted for fifty images and the success rate
achieved was 80%.
Keywords— Red blood cells, Morphology, Houghman
Transform, Watershed, Extreme points.
I. INTRODUCTION
certain RBCs change their size, shape, volume, structure or
color and become an abnormal cell. Such abnormalities are
detected by viewing the blood-smear images through a
microscope, which is a time consuming and error-prone
process. Thus there is a need for automation ofanalysis of
individual red cells in the blood smear image and segmenta-
tion of such cells using appropriate techniques. There are
various segmentation techniques that are used to segment all
types of cells in the blood smear images. These vary from
simple intensity based segmentation to fuzzy, neural net-
work or Tissue-like P Systems based segmentation. Intensi-
ty based segmentation was carried out to segment White
Blood Cells in the blood smear images [1]. A Marker Con-
trolled Watershed Transform was used to segment the nuc-
lei of WBCs [2] and a Distance Transform based Watershed
Transform [3] was used to segment the RBCs. Splitting of
overlapping cells was possible to a greater extent when
Watershed Transform was used [4] . In segmentation me-
thods based on region growing, the homogeneity criterion is
achieved by using the gray value and standard deviation of
the regions. Certain segmentation methods use the snake
algorithm [5] and few others used Tissue-like P Systems
[6]. Fuzzy logic can also be applied to segment WBCs [7]
[8].Appropriate segmentation techniques were also used to
segment abnormal RBCs based on their size and shapes.
Feature extraction was used to decide the type of abnormal
RBCs [9].
The proposed study aims at a Morphology-based detec-
tion of abnormal red blood cells in peripheral blood smear
images, based on their sizes and shapes. The rest of the
paper is organized as Background in section II, Methodolo-
gy in section III, Results and Findings in section IV and
Conclusion in section V.

A peripheral blood smear image has a rich representation

of White Blood Cells (WBCs), Red Blood Cells (RBCs) and
Platelets. Red blood cells are the most abundant type of
blood cells delivering oxygen to body tissues, the count of
which is often the first step in analyzing a patient’s patho-
logical condition. Due to various pathological disorders,

© Springer International Publishing Switzerland 2015 57

J. Goh and C.T. Lim (eds.), 7th World Congress on Bioengineering 2015
IFMBE Proceedings 52, DOI: 10.1007/978-3-319-19452-3_16
58
II. BACKGROUND
The four aspects in evaluating if a red blood cell is normal
or abnormal are its size, color, shape and inclusion bodies.
Normal RBCs are pink to reddish in color, 6-8 microns in
diameter and are round and biconcave disc shaped with high
surface to volume ratio, helping them to be flexible and
bendable as they pass through capillaries and allowing them
to absorb greater amounts of oxygen during their circulation
throughout the blood. Any variations in these characteristics
of RBCs are an abnormality. Few of such abnormal RBCs
based on their size and shape are discussed below:
x Anisocytosis: is a variation in size of RBCs, which
may be an indication of anemia [10].
x Microcytosis: is the abnormality where the size of
the red blood cells becomes less than 6 microme-
ters and has less amount of hemoglobin. This is
due to iron deficiency, defective hemoglobin, por-
phyrin synthesis, Thalassaemia, Sideroblastic ane-
mia, or Lead poisoning [10].
x Marocytosis: which are larger than normal RBCs,
is due to the occurrence of immature red blood
cells, vitamin B12 or folate deficiency and have a
blue-grey coloration [10].
x Ovalocytes: are oval shaped red blood cells, which
are due to Thalassaemia major and is hereditary.
x Elliptocytosis: is a disorder passed down through
families in which the red blood cells are abnormal-
ly shaped. Elliptocytosis affects about 1 in every
2,500 people of northern European heritage. It is
more common in people of African and Mediterra-
nean descent. The symptoms arefatigue, shortness
of breath and yellow skin and eyes (jaundice) [10].
x Codocytes: are nucleated RBCs, which are imma-
ture red blood cells, the presence of which indicate
accelerated erythropoiesis or severe bone marrow
stress in an adult [10].
x Acanthocytes and Echinocytosis: are spheroid
RBCs with a few large spiny projections. This can
be seen in patients with alcoholic cirrhosis, who
have inability to fully absorb dietary fats through
the intestines or severe liver disease [10].
x Dacrocytes: are RBCs with the shape of a tear
drop.These abnormal cells are due to severe iron
deficiency, Thalassemia major, bone marrow fibro-
sis orMegaloblastic anemia [10].
x In rouleax formation, the RBCs are arranged in the
form of a coin stack. It is due to the increase in the
blood concentration of fibrinogen, globulin and pa-
raprotein [10].
IFMBE Proceedings Vol. 52
S. Kulasekaran et al.
x In sickle anemia the RBCs are sickle shaped. This
is a Hb-S disease [10] [11].
In order to analyze these abnormal cells and diagnose
various disorders segmentation of such cells is necessary.
III. METHODLOGY
A. Image Pre-processing
The test images are first converted to binary images us-
ing Otsu’s thresholding. The objects that are touching the
borders are eliminated and holes filled in them. Morpholog-
ical operations like dilation and erosion are applied on to all
the blobs in order to smoothen them and strengthen their
borders [12][13]. The WBCs are eliminated based on their
sizes as they are visibly larger than RBCs. Detection of
abnormal cells are discussed in the following sub-section.
B. Detection of Abnormal Red Blood Cells
Microcytes and Polychronasisis are detected by analyz-
ing the size of the RBCS.The elliptical and oval shaped
RBCs, which are Elliptocytes and Ovalocytes respectively
are identified using Houghman Transform [14]. It is seen in
codocytes, which are nucleated RBCs, after segmentation
the holes in those cells are visible even after pre-processing.
Acanthocytes & Echinocytosis are identified by generating
a convex hull for each of the RBCS and analyzing the dis-
tance between the points in the convex hulls. In order to
detect the cells with the rouleax formation, a distance trans-
form based watershed algorithm is applied to the individual
stacks so that they split. The split cells are then counted and
if the count is above a particular value it is inferred that
such RBCs have Rouleax formation. In sickle anemia the
RBCs are sickle shaped. These sickle shaped cells are de-
tected using the roundness factor of blobs. The tear
dropped RBCs called dacrocytes are detected by obtaining
the extreme points of each of the cell.
IV. RESULTS AND FINDINGS
A prototype application was developed using MATLAB
for testingthe images used for the study. 50 blood smear
images with various RBC abnormalities were tested. The
images were first converted to grey images and necessary
algorithms were applied to them and the results were ob-
tained. A sample set of images, which were used for testing,
their pre-processed binary images and the resultant seg-
mented images of the abnormal cells are shown in Fig. 1,
Fig. 2, Fig.3 and Fig.4.