Image Stitching Using Scale Invariant Feature Transform and Counting of

Red Blood Cells and White Blood Cells using Canny Edge Detection
and Circular Hough Transform in Urine Sample
Ramon G. Garcia#1, Jennifer C. Dela Cruz#2
Mikko Ivan D. Avilledo#3, John Christopher M. Buera #4, Rom Vincent S. Chan#5, Paul Gian T. España#6
School of Electrical, Electronics, and Computer Engineering
Mapúa University
Manila, Philippines
1rggarcia@mapua.edu.ph 2jennycdc69@gmail.com

3mikkavilledo@yahoo.com 4chris.buera@gmail.com 5pg_espana@yahoo.com 6romvincentschan@yahoo.com

Abstract— Nowadays, counting of red blood cells (RBC) and
white blood cells (WBC) in a urine sample is done manually
by a medical technologist through use of the naked eye.
However, this makes the blood cell count subjective to the
skill of the lab technician and will take much longer time in
doing the task. This paper proposes the use of image
processing in counting the RBCs and WBCs in a urine
sample through use of various algorithms. The process of
consists of three (3) main parts. First is the autofocusing
using the variance of Laplacian method. Next is the use of
Scale Invariant Feature Transform (SIFT) as the image
stitching algorithm. Last is the automatic detection and
counting of blood cells using the Canny Edge Detection and
Circular Hough Transform algorithms. This study aims to
provide faster blood count results with accuracy as the
manual counting or even better.
Keywords— Microscopic, Urine, RBC, WBC, SIFT,
Canny Edge Detection, Circular Hough, Image Stitching
I. INTRODUCTION
In the field of biomedical science, urinalysis plays a basic
part in terms of health diagnosis. It is a laboratory test of urine
that can assess presence of blood in urine (hematuria) through
the sediment microscopic examination. One way of conducting
the hematuria test is by simply using a microscope to evaluate
presence of blood on the urine sample – manually counting the
number of red blood cells (RBC) and white blood cells (WBC)
present on the sample [1]. Manual counting is not reliable
seeing that this is subjective to the skill of a lab technician
performing the action [2]. To solve this problem, an image
processing method in RBC and WBC counting is proposed. On
these terms, immediate results can be expected.
Difficulty in counting the blood cells automatically is likely
to happen and this is due to its complex nature in segmentation.
Various methods from past studies that address the same issue
already existed [3] [4]. Using K-mean clustering was once
applied for segmentation purposes on an enhanced color image
[5]. Another is the use of the Circular Hough Transform which
has a feature of detecting the boundaries of the circles;
applying it to cells. The use of dynamic appearance model such
as circular shape counter has been applied in RBC
segmentation before [6].
Based from the previous studies using microscopic
images in analysing urine, researchers focused on the detection
of red blood cells in urine image captured under microscope by
image processing [6] [7]. From the study of Xu Ma, Gang An,
and Bin Li, it was found out that autofocus feature was not
applied [8]. Compared to this study, the variance of Laplacian
method will be used for autofocusing of the microscope. Image
stitching will also be applied using the Scale Invariant Feature
Transform algorithm [9]. As for the work of Navea, Dupo and
Bacudio image stitching was not considered. Also, blob
detection algorithm was used for the cell detection and
counting [5]. Combination of Canny Edge Detection and
Circular Hough Transform on the other hand will be used for
this study.
The main objective of this to count and display the number
of blood cells present in a urine sample using microscope
image processing Specific objectives are: (a) To create a device
that will capture segmented images of the urine sample using
varisance of Laplacian method for auto-focus feature, (b) To
use Scale Invariant Feature Transform for image stitching, (c)
To use Canny Edge Detection and Circular Hough Transform
as algorithms for detection and automatic counting of blood
cells, and (d) To test the accuracy of the device by comparing
its output with the manual testing results from laboratories.
This study will be beneficial to the field of medicine
particularly in urinalysis. The classical method of detecting
disease using urinalysis is by manual counting of red blood
cells, and white blood cells [3]. Knowing the presence of the
blood cells can determine if a patient has urinary tract infection
or kidney stones [10]. For the manual counting and focusing, it
will be subjective to the skill of the technician doing the task
unlike in automatic counting and focusing which is consistent
all throughout the process; thus, making it a scientific
approach. For the image stitching, it will remove the
overlapping which may improve the output accuracy.
Moreover, faster results will be obtained.
The scope of this study is it only focuses on the detection
and counting of blood cells, RBC and WBC, which are present
in a pre-centrifuged urine sample. Other urine content such as
bacteria, casts, epithelial cells and fat are not considered by the
device. Hence, there’s a probability of UTI and kidney stone
are the only diseases to be identified by the process. The result
of this device will be stored in cloud storage.
II. METHODOLOGY
The study makes use of pre-centrifuged urine sample as the
input to the image processing algorithms such as Variance of
Laplacian, Scale Invariant Feature Transform, Canny Edge
Detection, and Circular Hough Transform. Upon undergoing
these processes, the result shall be the count of detected blood
cells, both red blood cells (RBC) and white blood circles
(WBC), present in the urine sample.
Figure 1. Conceptual Framework of Methodology
point will be done. The captured image in this process is for the
auto-focus feature only. This method works to measure the
second derivative of an image. The Laplacian highlights
regions of an image containing rapid intensity changes.
Likewise, the method is often used for edge detection.
The assumption here is that if an image contains high
variance then there is a wide spread of responses, both edge-
like and non-edge like, representative of a normal, in-focus
image. But if there is very low variance, then there is a tiny
spread of responses, indicating there are very little edges in the
image. The more an image is blurred, the less edges there are.
To meet this objective, sixty captured images from
different altitudes of microscope’s tube, through use of a DC
gear motor connected to the coarse adjustment knob, will be
used. The variance of Laplacian of the image samples will be
computed, and the altitude of the tube where the sample image
has the highest value will be used for the next process –
automated microscope slide adjuster with image capture.

The processes mainly make use of the Raspberry Pi,

Raspberry Pi Camera Module, DC gear motor, and stepper
motors for image acquisition and image pre-processing.

A. Urine Sample Preparation and Image Acquisition

The process of preparing the urine samples and image
acquisition are as follows: (1) A clean (sterile) specimen bottle
will be provided to the respondents, (2) The respondents will
be asked to urinate in a specimen bottle, (3) A professional
medical technologist will perform a reagent strip test on the
sample, (4) The sample will then be centrifuged by the medical
technologist, preparing it for the microscopic urinalysis, (5)
After an average of five minutes, the sediments will be poured
in a sterile glass slide for the microscopic viewing, (6) The
sediments will be poured in a sterile glass slide for the Figure 3. Steps on Auto-focusing Process
microscopic viewing, (7) The glass slide will then be placed at
an exact location on the microscope stage which has a mark as
to where the slide must be properly positioned and (8) Capture C. Slide Adjusting and Image Capture
the image using a raspberry pi 3 camera module v2.1 placed in The slide adjustment and image capture is done through use
front of the eyepiece of the microscope. The camera setting is of two stepper motors connected to x- and y-axes of the slide
at 50% magnification while the eyepiece has 40x objective lens adjustment knobs of the compound microscope. Every capture
and 10x magnification – producing high power magnification from the Raspberry Pi camera module, the servo motors
of 400x. attached to the knobs will act in an interval manner. The
images captured here will serve as the input of the next process
which is the image stitching.

Figure 2. Sample Preparation and Image Aquisition

B. Variance of Laplacian Method for Auto-Focus Feature

The variance of Laplacian method is used as an Figure 4. Slide Adjustment Process
autofocusing technique wherein finding the captured image
with the highest variance and setting the focus to that certain

D. Scale Invariant Feature Transform (SIFT) for Image
Stitching
Scale Invariant Feature Transform will be used for image
stitching. In SIFT, it works very well even under rotation and
scaling changes between images when matched. SIFT is much
efficient compared to other algorithms in image stitching since
its features are robust to occlusion. It has a disadvantage which
is the ghosting, but it only occurs if the subject is moving.
Since our subject is only at stand still it won’t be possible to
have a ghosting.
 From the image formed from the last procedure for the
canny edge, this algorithm will detect the blood cells in
which they are circles in the image.
 Once the circle/s is identified, the center and the radius
will be determined.
 Based from the radii range, the circles within the given
range will be found by the algorithm and they will be
specified as a scalar of any numeric type.
 It will be counted as a red blood cell if the radius is
within the 3.5 to 4.25 micrometer range. When the
radius is within 5 to 7.5 micrometer, the contour will be
counted as a white blood cell.

III. RESULTS AND DISCUSSION

Distance Trial 1 Trial 2 Trial3 Actual

record
1
2
3
4
5
6
7
8
9
Figure 5. SIFT Image Stitching Processes
10

E. Combination of Canny Edge Detection and Circular Hough

Transform Algorithms for Detection and Counting of Blood
Cells
The Canny Edge Detection is the algorithm that will be
used for rescaling the image into grayscale and forms the edges
of the image. The Canny Edge Detection will be used in the
rescaling part since it is less susceptible of noise compared to
any other edge detection algorithms and this algorithm is suited
Sample Blood Compare Monitor
for low quality images. The Circular Hough Transform is the
Worker pressure the result worker
one responsible in detecting the blood present in the urine.
reading from with Status
MATLAB will be the program used with inputs of image,
PC standard or Send
radius range, sensitivity factor, and edge threshold. Once these
(Thingspeak) diagnostic a
algorithms were implemented, the output will then be the count
values medical
of the detected blood cells as displayed from a graphical user
team
interface (GUI).

Canny Edge Detection

 The algorithm will check its current gradient and
magnitude.
 Non-maxima suppression will examine the pixel for
information.
 A high and low threshold will be set to determine which
part will be removed from the image.
 Hysteresis is the one responsible to detect the strong and
weak edges based from the threshold.

Circular Hough Transform

 A range of radii will be set. A smaller range will lead to
higher accuracy.
 Circular Hough Transform to determine the number of red
blood cells (RBC) and white blood cells (WBC). Furthermore,
the researchers have tested the accuracy of the device base on
Wor Tri Tri Tri Tri Tri Tri Tri Tri Tri Tri Res
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the manual blood cell count by determining the mean of error
1 2 3 4 5 6 7 8 9 10 for each type of blood cell. The RBC mean of error (%)
produced a higher error compared to that of the WBC.
Nevertheless, minimal percentages of error were obtained for
the both blood cells.

V.REFERENCES
[1] Tangsuksant, W., Pintavirooj, C., Taertulakarn, S., &
Daochai, S. (2013, October). Development algorithm to count
blood cells in urine sediment using ANN and hough transform.
In Biomedical Engineering International Conference
(BMEiCON), 2013 6th (pp. 1-4). IEEE.

[2] Ma, X., An, G., & Li, B. (2009, May). Design and
Implementation of an Automated Microscope Stage. In
Information Technology and Applications, 2009. IFITA'09.
International Forum on (Vol. 1, pp. 603-605). IEEE.

[3] Bernadette F.. Rodak, George A.. Fritsma, &

Doig, K. (2007). Hematology: Clinical
Principles and Applications. WB Saunders.

[4] Tworek, J. A., Wilkinson, D. S., & Walsh, M. K. (2008).

The Rate of Manual Microscopic Examination of Urine
Sediment: A College of American Pathologists Q-Probes Study
of 11 243 Urinalysis Tests From 88 Institutions. Archives of
pathology & laboratory medicine, 132(12), 1868-1873.

[5] Navea, R. F., Dupo, V., & Bacudio, L. (2015). Red Blood
Cells and White Blood Cells Detection, Differentiation and
Counting using Image Processing.
Figure 7. Urine Sample After WBC Detection [6] Lin, Z., Yu, Y., & Chen, T. (2012, March). An auto-focus
algorithm based on image processing. In Automatic Control
For this part, comparison between the counts using and Artificial Intelligence (ACAI 2012), International
manual counting versus the automatic counting is exhibited Conference on (pp. 394-397). IET.
on Table 2. Subsequently, the percentage error is shown on
Table 3 using the percentage error formula that is defined Trial T(sec)
as 1
2
𝑀𝑎𝑛𝑢𝑎𝑙 𝐶𝑜𝑢𝑛𝑡−𝐴𝑢𝑡𝑜𝑚𝑎𝑡𝑖𝑐 𝐶𝑜𝑢𝑛𝑡
% 𝐸𝑟𝑟𝑜𝑟 = |
𝑀𝑎𝑛𝑢𝑎𝑙 𝐶𝑜𝑢𝑛𝑡
| 𝑥 100 3
4
The average percentage error (Mean of Error %) of 5
RBC and WBC counting are 5.28 and 9.65, respectively.
6
TABLE 2. Comparison between manual and automatic 7
blood cell counting 8
9
10

TABLE 3. Average percentage error of RBC and WBC
counting
IV. CONCLUSION
The study has successfully developed a device that uses a
Raspberry Pi Camera for image acquisition and performs the
image processing algorithms such as Variance of Laplacian,
Scale Invariant Feature Transform, Canny Edge Detection, and
[7] Obara, T., Igarashi, Y., & Hashimoto, K. (2011,
September). Fast and adaptive auto-focusing algorithm for
microscopic cell observation. In Intelligent Robots and Systems
(IROS), 2011 IEEE/RSJ International Conference on (pp. 7-
12). IEEE.
[8] Zhu, Q., & Li, K. (2010, June). Image stitching using
simplified SIFT. In Information and Automation (ICIA), 2010
IEEE International Conference on (pp. 1134-1137). IEEE.
[9] Mazalan, S. M., Mahmood, N. H., & Razak, M. A. A.
(2013, December). Automated red blood cells counting in
peripheral blood smear image using circular Hough transform.
In Artificial Intelligence, Modelling and Simulation (AIMS),
2013 1st International Conference on (pp. 320-324). IEEE.

[10] Song, Y., Li, M., & Sun, L. (2006, December). A new
auto-focusing algorithm for optical microscope based
automated system. In Control, Automation, Robotics and
Vision, 2006. ICARCV'06. 9th International Conference on
(pp. 1-5). IEEE.