Microscopic Image Analysis and Counting of Red Blood
Cells and White Blood Cells in a Urine Sample
Jennifer C. Dela Cruz Ramon G. Garcia
School of EECE School of EECE
Mapua University Mapua University
jcdelacruz@mapua.edu.ph rggarcia@mapua.edu.ph
John Christopher M. Buera Rom Vincent S. Chan
School of EECE School of EECE
Mapua University Mapua University
chris.buera@gmail.com romvincentschan@yahoo.com
ABSTRACT
Traditional counting of red blood cells (RBC) and white blood
cells (WBC) in a urine sample is done manually by a medical
technologist. However, this makes the blood cell count subjective
to the skill of the laboratory technician and will take much longer
time in doing the task. This paper proposes the use of image
processing in counting the WBCs and RBCs in a urine sample
through use of Canny Edge Detection and Circular Hough
Transform algorithm. The process consists of two (2) main parts.
First is the Canny Edge Detection and the final part is Circular
Hough Transform algorithms. It shows that the proposed system
has a percentage accuracy of at least 93.229% in reference to the
actual RBC and WBC count result.
CCS Concepts
Computing methodologies ➝ Computer Graphics ➝ Image
Manipulation ➝ Image Processing
Keywords
Blood cell counter; Canny Edge Detection; Circular Hough;
Microscopic; RBC; Urine; WBC
1. 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. Manual counting is subjective to the skill
of a lab technician performing the action. This process is also
known to be tedious and repetitive. These factors may lead to
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© 2019 Association for Computing Machinery.
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https://doi.org/10.1145/3326172.3326185
113
Mikko Ivan D. Avilledo
School of EECE
Mapua University
mikkoavilledo@gmail.com
Paul Gian T. Espana
School of EECE
Mapua University
pg_espana@yahoo.com
incorrect count of blood cells in urine. To solve this problem, an
image processing method in RBC and WBC counting is proposed.
On these terms, even after many repetitions a consistent and
accurate counting of blood cells is done.
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. Using K-mean clustering was once applied for
segmentation purposes on an enhanced color image. 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.
The main objective of this research is to count and display the
number of blood cells present in a stitched image of urine sample
using microscope image processing. Specific objectives are as
follows: (a) To use Canny Edge Detection and Circular Hough
Transform as algorithms for detection and automatic counting of
red blood cells (RBC) and white blood cells (WBC), and (b) To
test the accuracy of the device by comparing its output with the
manual testing results from laboratories.
This study aims to be beneficial to the field of medicine
particularly in urinalysis. Knowing the presence of blood cell in a
urine sample can help determine if a patient has a urinary tract
infection (UTI) or kidney stones. The classical method of
detecting disease using urinalysis is by manually counting red
blood cells and white blood cells through microscopic analysis.
And for the manual focusing and counting, it is subjective to the
skill of the technician doing the task unlike in automatic counting
which is constant all throughout the process; thus, making it a
scientific approach.
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. The
result of this device will just be available at the time the device
was used; thus, no data storage feature is included.
2. REVIEW OF RELATED LITERATURE
This paper proposes microscopic analysis of blood cells present in
a urine sample through integration of various features namely
auto-focus, image stitching, and automatic detection and count of
blood cells. In this chapter, different research papers that share the
same features as mentioned above are presented.
Based from the previous studies using microscopic images in examined with the use of microscope with a magnification of
analyzing urine, researchers focused on the detection of red blood 400x equipped with Motic camera for capturing.
cells in urine image captured under microscope by image
processing. From the study of Xu Ma, Gang An, and Bin Li [1], it 3. METHODOLOGY
was found out that autofocus feature was not applied. Compared The study makes use of pre-centrifuged urine sample as the
to this study, the variance of Laplacian method will be used for subject to the image processing algorithms namely Canny Edge
autofocusing of the microscope.In this paper of Navea, Dupo, and Detection, and Circular Hough Transform. Upon undergoing these
Bacudio arises as closest to this study. According to their paper, a processes, the result shall be the count of detected blood cells,
blob detection algorithm was used to detect and differentiate both red blood cell (RBCs) and white blood cells (WBCs) present
RBCs from WBCs [1]. Another technique that was used from in the urine sample. Please refer to Figure 2.
other paper is the Hue, Saturation, Value (HSV) thresholding
method and connected component labeling [2]. According to the
paper of Dela Cruz, et al., HSV thresholding background
subtraction was used for blood cell segmentation while connected
component labelling was used for blood cell counting. The study
of Tangsuksant, et al. proposed the segmentation and detection of
RBCs and WBCs in urine sediment images [3]. This is through
use of development algorithm to count blood cells in urine
sediment using ANN and Hough Transform. From the study of
Dela Cruz, et al., the researchers determined blood components
(WBCs, RBCs, and Platelets) count in microscopic images using
image processing and analysis [4]-[6]. Here, the sample used for
the image processing is a stained blood. The process of classifying Figure 2. Conceptual framework of methodology
and placing sharp discontinuities in an image is called the edge
detection [7].
The canny edge detection algorithm also known as optimal edge
detector is used for medical and defense application. Also, in
industry, the Canny edge detection technique is one of the
standard edge detection techniques. The algorithmic steps are as
follows as can be seen in Figure 1.
• Convolve image f(r, c) with a Gaussian function to get
smooth image f^(r, c). f^(r, c)=f(r,c)*G(r,c,6)
• Apply first difference gradient operator to compute edge
strength then edge magnitude and direction are obtained as
before.
• Apply non-maximal or critical suppression to the gradient
magnitude
• Apply threshold to the non-maximal suppression image.

Figure 3. Canny edge detection flowchart

The processes were employed through use of the Raspberry Pi,

Figure 1. Implementation of Canny Edge Detection on a cell
sample
The study done by Mazalan, Mahmood and Razak is about
counting of red blood cells with the use of Circular Hough
Transform [8]. From their research, a blood was sampled and was
114
Raspberry Pi Camera Module, image pre-processing. The Canny
Edge Detection is the algorithm used for rescaling the image into
grayscale and forms the edges of the image. Please refer to
Figures 3 and 4. It is used for the rescaling part since it is less
susceptible of noise compared to any other edge detection
algorithms. On the other hand, the Circular Hough Transform is
the one responsible in detecting the blood present in the urine.
Python and OpenCV are the software used with stitched image,
radius range, sensitivity factor, and edge threshold as the input. The Circular Hough Transform is the algorithm used for detection
Once these algorithms were implemented, the output will then be and counting of the Red blood cells and White blood cells. A
the count of the detected blood cells as displayed from a graphical range of radii will be set. From the image formed from the last
user interface (GUI). procedure for the canny edge, this algorithm detects and counts
the blood cells and encircles them with red circles (RBCs) and
1 2a 2b green circles (WBCs). (Please see Figures 5-6 for the flowchart
and sample images) Once the circles are 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 2 to 4-pixel
2c 3 4 range. When the radius is within 4.5 to 15 pixels, the contour will
be counted as a white blood cell.

Figure 4. Implementation of Canny Edge Detection: (1)

Gaussian Smoothing, (2a) Gradient (magnitude), (2b)
Gradient in X, (2c) Gradient in Y, (3) Non-maximum
suppression, and (4) Hysteresis Threshold
Figure 6. Implementation of Circular Hough Transform;
RBCs enclosed in red circles, and WBCs enclosed in green
circles
For automatic blood cells counting, the automatic blood cell count
for RBCs and WBCs are obtained by using equations 1 and 2.
Here, actual blood cell count is equated to the total number of
detected blood cells divided by the number of fields considered
for the process of SIFT image stitching. These numerical values
are then to be compared with the manual blood cells counting. For
automatic blood cell counts:

(1)

(2)

4. RESULTS AND DISCUSSION

The comparison between manual counting versus the automatic
counting of RBCs and WBCs is shown in Table 1. For automatic
blood cells counting, the automatic blood cells actual count for
RBCs and WBCs are obtained by dividing the total number of
detected blood cells by the number of images stitched in the
process of SIFT image stitching. These numerical values are to be
compared with the manual blood cells counting.
Test comparisons for manual RBC and automatic RBC counts are
shown in Figure 7. Test comparisons for manual WBC and
automatic WBC counts are shown in Figure 8.

Figure 5. Circular Hough Transform Flowchart

115
 Table 1. Comparison between manual and automatic blood cell count
Number of Fields considered for Manual Counting: 88
Number of Fields considered for Automatic Counting: 32
Manual Blood Cell Count Automatic Blood Cell Count
Automatic Blood
Actual Blood Cell Total Blood Total Detected
Urine Sample Cell Actual
Count Cell Count Blood Cell Count
Count
RBC WBC RBC WBC RBC WBC RBC WBC
1 6 10 48 80 185 324 5.78 10.13
2 1 6 8 48 33 191 1.03 5.97
3 4 29 32 232 126 924 3.94 28.88
4 2 6 16 48 63 193 1.97 6.03
5 0 1 0 8 2 33 0.06 1.03
6 3 16 24 128 93 505 2.91 15.78
7 9 56 72 448 288 1804 9 56.38
8 5 9 40 72 159 281 4.97 8.78
9 7 12 56 96 225 375 7.03 11.72
10 9 9 72 72 286 282 8.94 8.81

The percentage errors for both RBCs and WBCs are shown on
Table 2 using the percentage error formula as defined in (3).

(3)
The average percentage errors (Mean of Error %) for RBC and
WBC counts are 1.56 and 1.06, respectively.

Table 2. Average percentage error of actual RBC and WBC

count
Urine Sample % Error
Figure 7. Test comparisons for Manual RBC count VS. RBC WBC
Automatic RBC Count 1 3.67 1.30
2 3.00 0.50
3 1.50 0.41
4 1.50 0.50
5 0 3.57
6 3.00 1.38
7 0.00 0.68
8 0.60 2.44
9 0.43 2.33
10 0.67 2.11
Mean of Error (%) 1.55 1.06
Figure 8. Test comparisons for Manual WBC count VS.
Automatic WBC Count

116
 Table 3. RBC actual count result and WBC count results have no significant difference in
comparison to the manual RBC and WBC count results because
Urine Automatic Manual
d the absolute values of the calculated Ts for both RBC and WBC
Sample Count Count
are less than the t-table at an alpha level of 0.05.
1 5.78 6 -0.22
Table 5. Normal range of RBC and WBC in a urine sample
2 1.03 1 0.03
Parameter Normal Range
3 3.94 4 -0.06
Red Blood Cell (RBC) 0 – 2 RBCs/hpf
4 1.97 2 -0.03
White Blood Cell (WBC) 0 – 5 WBCs/hpf
5 0 0 0
6 2.91 3 -0.09
Indicated above in Table 5, is the normal range of red blood cells
7 9 9 0
(RBCs) and white blood cells (RBCs) in a human urine sample.
8 4.97 5 -0.03
9 7.03 7 0.03
5. CONCLUSIONS
The study has successfully integrated image processing
10 8.94 9 -0.06 algorithms that allows determination, differentiation, and counting
Mean of differences -0.0446 of red blood cells (RBCs) and white blood cells (WBCs) present
in a urine sample. This research employed Canny Edge Detection
Standard deviation of differences 0.137 and Circular Hough Transform using Raspberry Pi 3 Model B.
t -1.778 These were used for the detection and counting of the WBCs
present in a urine sample. For the output, the final counts are
displayed on a GUI made. The result from the automatic blood
Table 4. WBC actual count result cell count were compared to the manual blood cell count on the
same urine samples, and this was made possible with the help of
Urine Automatic Manual
d the partner laboratory. Furthermore, the researchers have tested
Sample Count Count
the accuracy of the device base on the manual blood cell count by
1 10.13 10 0.13 determining the mean of error for white blood cell and red blood
2 5.97 6 -0.03 cell. The RBC count mean of error (1.55%) produced a higher
error compared to that of the WBC count (1.06%). With this,
3 28.88 29 -0.12 using a sample size of thirty (30), the proposed system has a
4 6.03 6 0.03 percentage accuracy range of at least 93.229% with respect to the
manual blood cell count administered by the partner laboratory.
5 0.97 1 -0.03
6 15.78 16 -0.22 6. RECOMMENDATIONS
It is recommended to improve the accuracy of the proposed
7 56.38 56 0.38 system. Other image processing algorithms can be applied, and
8 8.78 9 -0.22 more urine samples can be examined to verify the effectivity of
the system. Future researchers are also advised to include a
9 11.72 12 -0.28
system data storage so that data retrieval will be easier. Finally, it
10 8.81 9 -0.19 is recommended to count other urine components such as crystals
Mean of differences 0.0497 and bacteria.

Standard deviation of differences 0.234 7. REFERENCES

t 1.278
The average percentage error of actual RBC and WBC are shown
in Tables 2-4.
The null and alternative hypotheses for t-tests shown above are as
follows:
• Null Hypothesis: There is no significant difference on the
manual RBC and WBC count results of the urine samples
tested, with that of the proposed automatic count.
• Alternative Hypothesis: There is significant difference on the
manual RBC and WBC count results of the urine samples
tested, with that of the proposed automatic count.
Through use of the t-distribution table, with 2-tail confidence
level of 95% and alpha (significance) level of 5%, t-values within
the range -2.2622 to +2.2622 are accepted. Given the data
presented above, it was proven that the proposed automatic RBC
117
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