Spermatozoa Morphology Examination Using LenshookeTM SQA
X1 Pro Compared with Manual Method
Adhipireno Purwanto1, Purwanto Edy2,3, Suyono Seso Sulijaya4, Ismawatie
Emma1,5, Santoso Budi6, Limijadi Edward Kurnia Setiawan3
1

Clinical Pathology Laboratory Department
of Central General Hospital Dr. Kariadi
Semarang, Semarang, Central Jawa,
Indonesia
2

Master Program of Medical Laboratory
Science/Clinical, Faculty of Nursing and
Health Sciences, University of
Muhammadiyah Semarang, Semarang,
Central Jawa, Indonesia
3

Islamic Hospital Fatimah Cilacap, Central
Jawa, Indonesia
4

Gladiool IVF Centre, Magelang city,
Central Jawa, Indonesia
5

Clinical Pathology Laboratory Department
of Mother and Child Hospital Restu Ibu
Sragen, Central Jawa, Indonesia
6

Muhammadiyah Semarang University,
Central Jawa, Indonesia
Correspondence:
Purwanto Edy,
Jl. Kedungmundu No.18, Kedungmundu,
Kec. Tembalang, Kota Semarang, Semarang,
Central Java
Zip Code: 50273
Email: ichtiar9@yahoo.co.id
Received: April 17, 2021
Revised: Januari 1, 2022
Accepted: February 22, 2022
Published: April 28, 2022
DOI: 10.33086/ijmlst.v4i1.2059

Abstract
According to World Health Organization data, 30-40% of
infertility is caused by male factors. The morphology of
normal spermatozoa is an indicator of male fertility, and it
is known by manual or automatic sperm analysis.
LenshookeTM SQA X1 PRO automatic equipment comes
along with the development of laboratory equipment
automation technology. The working principle of this tool is
by shining light on the object of examination, then the
camera with high resolution, with the facility of an optical
lens will take a picture of the object. The database recorded
by the camera is analyzed by the algorithm. The research
objective was to test the suitability of the Lenshooke TM SQA
X1 PRO automatic tool with manual method as the Gold
Standard. Subjects in this study were patients who carried
out semen analysis tests at the Clinical Pathology
Laboratory of RSIA "Restu Ibu" Sragen from June to
August 2020. The examination method used an automatic
method with the LenshookeTM SQA X1 PRO tool and a
manual method with Papanicolaou staining. The results of
the study, conformity test with WHO 2010 normal
standards, automatic methods reached 94.4% compared to
manual methods. The next statistical test was with standard
mean, normal sperm morphology data had a significance of
0.001, abnormal sperm head data had a significance value of
0.956 and abnormal sperm tail data had a significance value
of 0.339. The LenshookeTM SQA XI PRO device based on
automatic technology can be used in laboratory services for
sperm analysis in addition to manual methods. Suggestions
for using the LenshookeTM XI PRO automatic tools are still
accompanied by the manual method.
Keywords
Automatic, Microscope, Morphology, Spermatozoa.

This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use,
distribution, and reproduction in any medium, provided the original work is properly cited. ©2021 by author.

45

Adhipireno Purwanto, et all.

Morphological abnormalities can make it

INTRODUCTION
The morphology of spermatozoa refers

difficult for spermatozoa to fertilize an egg

to the whole form of spermatozoa cells

(6).

Spermatozoa

consisting of the head, middle and tail (1).

variability in size and shape. There is a

Spermatozoa cells form in the seminal

correlation between the morphology of

tubules that are in the testes. This tubule

spermatozoa

contains a complex series of cells, namely the

reproductive tract during the fertilization

development or division of cells from the

process (7).

and

show

reaching

tremendous

the

female

germinal to the formation of spermatozoa or

Analysis of semen in manual method is

male gametes (2). About 300 million

the gold standard (WHO), which is widely

spermatozoa carry out the process of

used in laboratory of Clinical Pathology. This

spermatogenesis mean in one day there are

method has the advantage and disadvantage.

approximately 300 million

Some of the findings in the field for the

spermatozoa

shortcomings of the manual method of semen

newly manufactured (3).
According to data from the World Health

analysis include that there are still differences

Organization (WHO), 30 – 40% of infertility

in the interpretation of the results of inter

is caused by male factors, hence it is

laboratory

important to evaluate fertility of men as part

process takes a relatively long time between

of routine check. The basic test for infertility

30 to 60 minutes, and the equipment used

by performing semen analysis is the most

does not have the same standard (4).

commonly used diagnostic option. The result

The

examinations.

morphological

The

analysis

examination

of

of semen analysis of 25% of infertile men

spermatozoa using an automatic method is

was asthenozoospermia (abnormality of

present amidst the need for semen analysis.

spermatozoa movement). The rest are

The development of accessible, fast and

disturbances in number (oligozoospermia)

standard methods for semen analysis is

and morphology (teratozoospermia) or a

urgently needed. The automated method

combination of the three (4).

provides a solution for semen analysis checks

Semen analysis includes macroscopic

with fast results and good quality control

and microscopic examinations. Microscopic

standards

analysis of normal semen results from

automated with computer-based method to

parameters of concentration, motility and

cover the shortage of existing shortcomings

morphology to determine fertility in men.

in the semen analysis manual methods (8).

Normal spermatozoa cell morphology as a
clinical

tool

for

male

fertility

(5).

(4).

Semen

analysis

using

The basis for the authors’ consideration
as Medical Laboratory Technologist Experts
59

Adhipireno Purwanto, et all.

in conducting research entitled Spermatozoa
Morphological
Lenshooke

TM

Examination

Using

the

SQA X1 PRO Tool Compared

MATERIALS AND METHODS
Morphology
grouped

into

of

spermatozoa

normal

and

were

abnormal

to the Manual Method is because the authors

morphology (5). Sampling was performed by

still find that there are differences in the

using a consecutive sampling, that is

interpretation of semen analysis results

sampling technique to assign subjects that

between one laboratory and another. Some

met the study criteria. Samples were taken

literature also states that manual semen

from patients who visited the Clinical

analysis is a simple and inexpensive test, but

Pathology Laboratory of Restu Ibu Hospital,

has high variability and is very subjective (9).

Sragen, Indonesia from June 2020 to August

Automatic
spemartozoa
Lenshooke

method
analysis

TM

selections

for

2020. The number of samples in this study

may

the

was 48 patients, which are consist of the 36

use

SQA X1 PRO. This tool

patients who met the criteria for the study.

consists of software and hardware and the

The inclusion criteria in this study were

technical analysis works automatically (10).

men in reproductive age between 20 to 45

This is very practical and simple tool, which

years

has four key parameters for evaluating male

concentration in excess of 2 million/mL.

fertility,

Exclusion criteria were semen samples with

namely

morphological,
Lenshooke

TM

concentration,

motility

and

pH.

SQA X1 PRO yields very fast

old

and

with

spermatozoa

blood mixture, abstinence more than 7 days,
liquefaction

more

than

60

minutes,

examination results which only take 3 to 5

increasing number of leukocyte in semen,

minutes to get all the test results. Good

and the number of immature spermatozoa

quality control would be giving results in a

cells.

accurate and reliable (11).

Sperm Fluid Release

As

a

conducted
examination

comparison,
a

the

manual
of

researchers

The release of sperm fluid for good

morphological

results is by masturbating without using

spermatozoa

using

tools, such as gels, detergents and others.

Papanicolaou stain because this is one of the

Sampling through sexual intercourse is not

WHO Gold Standards for spermatozoa

recommended. If circumstances compel

morphological staining (12). The aim of this

sampling by sexual intercourse then use

study was to evaluate the spermatozoa

condoms and lubricants that are non-toxic

morphology using Lenshooke

TM

and fertility-friendly if necessary. Collect

compared with manual method in order to

complete sperm fluid, especially the first

provides reliable results.

fraction rich in sperm.

Ina. J. Med. Lab. Sci. Tech. 2022; 4(1): 45–59

47

SQA X1 Pro

Adhipireno Purwanto, et all.

Sample Handling

staining has been proven and recommended

Prepare two sample pots, A and B, each

by the WHO (12). Polychromatic staining is

with a patient identification label. The sample

considered a very reliable staining technique.

is accommodated in a clean, dry and wide-

Factors that affect the coloring in addition to

mouthed sample pot. The research sample for

the use of dye solution, the time of painting,

one identity is divided into two in the sample

the duration of immersion, rinsing and

pot A and the sample pot B. Sample pot A for

immersion currents follow the standards that

inspection of the Lenshoke SQA X1 PRO

have been set. Figure 1 shows slides stained

automatic method tool. Sample pot B for

using the Papanicolaou procedure. This stain

manual

can be permanently installed and stored for

method

examination

with

Papanicolaou stain. Each sample pot is done
at the same time using two different

use as internal quality control (13).
The

principle

of

the

staining

of

inspection methods. To maintain the quality

Papanicolau The Harirs's haematokxylin dye

of the sample in the specimen pot, the

stains the cell nucleus blue, Orange G and EA

temperature is kept around 20 – 37°C.

50 alcohol-based green coloring will work to

Manual Method

color the cytoplasma. Ethanol 50% 80% 95%

In the manual method, Papanicolau dye

100% for fixation and make cells become

was used and considered that this dye is one

dehydrated and ethanol acid removes dyes

of the dyes recommended by WHO. Staining

undesirable but still attached especially to the

Papanicolaou gives the results of the

cytoplasmic area. Water rehydrates cells

examination both for the morphology of

(13). Procedure for manual method shows in

spermatozoa and other cells. Papanicolaou

Figure 1.

(A)

(B)

Figure 1. Preparation; 5-20 µL of semen is dripped on the object glass. (A) Move the slide to
another object to make an erase. (B) Dry in the air 5-15 minutes (WHO, 2010).
Papanicolaou coloring

hematoxylin stain for 4 minutes, then into

Soak the dried slides sequentially on

pure water 30 seconds, put into ethanol acid

ethanol 80% for 30 seconds, continue to

for 8 seconds, flux with cold tap water for 5

ethanol 50% 30 seconds and soak in pure

minutes, and then into alcohol 50% for

water for 30 second. Put into Harris

seconds, and 80% for 30 seconds, then dip
59

Adhipireno Purwanto, et all.

into ethanol 95% for 15 minutes. Stain with

green dye for 1 minute, and then rinse with

G-6 orange dye for 1 minute, and dip

ethanol 95% two times with 30 second each.

repeatedly into ethanol 95% for three times

Final clearing rinse with ethanol 100% two

with 30 seconds each. Stain with EA-50

times for 15 second each.

A

B

Figure 2. Morphology of spermatozoa with Papanicolau dye. (A) showing spermatozoa with
amorphous head with thickened midpiece. (B) round head (14).
Result Reading

according to defects in the head, neck,

Morphology reading is the one semen

midsection and tail (2). Several recent studies

analysis parameter in medical labotatory.The

have

demonstrated

the

importance

of

slides then read on a microscope with at least

assessing the morphology of abnormal

two technical officers. Using 1,000 times

spermatozoa more carefully to establish the

magnification assisted with immersion oil,

diagnosis of infertility (13).

observe the morphology of normal and
abnormal

spermatozoa.

Report

the

Head Normal

percentage of observations of spermatozoa

The head of the sperm cell is oval with a

morphology as a result of the study (Figure

size of 3 – 5 µm, there is a cell nucleus

2).

(nucleus) containing genetic information in
The

morphology

of

the form of DNA in it. This genetic

includes the assessment of the head, neck,

information will meet the genetic information

middle and tail. The normal form of

from the egg and will determine whether the

spermatozoa is a tadpole which consists of a

fetus is male or female. In the head of the

blunt head in which there is a nucleus, and

spermatozoa, there are also enzymes, such as

has a tail that contains an apparatus for

the hyaluronidase enzyme, which functions

moving.

to penetrate the corona layer above the ovum,

Morphologically

abnormal

spermatozoa are categorized into subgroups
Ina. J. Med. Lab. Sci. Tech. 2022; 4(1): 45–59

and protease enzymes (18).

49

spermatozoa

Adhipireno Purwanto, et all.

Neck Normal

cytoskeleton that functions to propel the

Neck is the area just behind the head that

spermatozoa forward, at a speed of 30

contains the centrioles. The middle part

inches/hour (18).

contains mitochondria arranged in a spiral,

Abnormal

which contains energy (ATP) as an energy

Abnormal is an abnormal form of

source for spermatozoa, for locomotion to the

spermatozoa.

site of fertilization and for spermatozoa

spermatozoa are categorized into subgroups

metabolism (3).

according to defects in the head, neck,

Tail

midsection and tail (1). Several recent studies

The tail of the spermatozoa is long with a

have

Morphologically

demonstrated

the

abnormal

importance

of

size of 50 µm divided into the neck, the

assessing the morphology of abnormal

main/middle and the end (19). The main part

spermatozoa more carefully to establish the

is the longest part of the tail, and the end is

diagnosis of infertility (13). Term the results

the pointed end of the tail. The tail of the

of semen analysis used to describe the

spermatozoa is in the form of flagella as a

morphological abnormalities of spermatozoa

means of locomotion in the form of a long

shows in Figure 3..

Figure 3. Abnormal spermatozoa morphology (5).

Head
Large or small, tapering, bulb-shaped /

Midpiece
Midpiece

defects

includes

tailless

pyriform, amorphous, hollow >20% of the

spermatozoa that appear as free heads, or

head area is occupied by a cavity that is not

loose

stained, the head is broad and a combination

swollen/irregular

central

of the above.

Abnormally

midsection

heads,

thin

uninserted

tails
bent
e.g.

or
tails.
no
59

Adhipireno Purwanto, et all.

mitochondrial

sheath

or

various

capturing images from microscopes, massive

combinations of these abnormalities.

increases in computing power along with

Tail

tremendous reductions in computer size, new

Short tails, double tails, shaped like
hairpins, broken, curled tails with drips at the
ends,

or

a

combination

of

these

computer languages, and updated software
algorithms (17).
In this study, the automatic tool used is

abnormalities.

the LenshookeTM SQA X1 PRO, a product

Automatic Method

from Bonraybio, a device that works

The tool used is the LenshookeTM SQA

automatically for human semen analysis by

X1 PRO, a product from Bonraybio, a device

integrating mechanical, optical, electronic

that works automatically for human semen

and algorithmic technologies.

analysis by integrating mechanical, optical,

The

working

principle

of

the

electronic and algorithmic technologies.

LenshookeTM SQA X1 PRO tool is with a

With the use of this Semen Quality Analyzer,

beam of light on the object of the

it will facilitate and improve performance in

examination, and then a high-resolution

the laboratory so that work is more efficient.

camera, with an optical lens facility, will take

Semen analysis using equipment equipped

pictures of the object. Furthermore, the

with a computer is an automated method that

clinical database that has been recorded by

can

the camera is analyzed for calculations with

provide

objective

and

precise

information about the characteristics of
semen

samples,

morphology,

The initial rare Work Procedure prepares

concentration, and motility (16). Quality

the sample by putting this sperm collection

Control of the Cement Quality Analyzer can

device for the Semen Quality Analyzer using

be standardized for each tool, so as to

a special consumable cup test, and then wait

minimize differences in the results of cement

30 to 60 minutes for sperm to thaw.

analysis

Semen

Homogenized the sample in the cup by

analysis in combination with computer

turning it back and forth 8 – 10 times, check

technology has evolved over the past 40

the color and volume of the sperm sample

years, through advances in devices for

(Figure 4-5).

between

such

as

the algorithm (15).

laboratories.

Ina. J. Med. Lab. Sci. Tech. 2022; 4(1): 45–59

51

Figure 4. How to operate the LenshookeTM SQA X1 PRO appliance (11).

Adhipireno Purwanto, et all.

Figure 5. Display results on the LenshookeTM SQA X1 PRO tool showed pH of the semen, total
spermatozoa in million per mililiters semen, total moved of the spermatozoa, and amount of the
total normal morphology of the spermatozoa (11).
Spermatozoa morphology parameters that

Abnormalities in sperm size and morphology

emerged from readings using the LenshokeTM

assessment includes the percentage of head

SQA

X1

PRO

device

were

normal

length, head width, head circumference, head

morphology

with

units

of

percent.

area and tail length (15).

RESULTS
Conformity Test

by the manual method, all showed normal

The suitability of the results of the
automatic

method

of

morphology

examination

spermatozoa

This

results

in

abnormal

morphological data of 0. The agreement that

manual

can be made with the 2010 WHO standard

method of spermatozoa morphology using

normal value ≥ 4% is by looking at the

the 2 x 2 Contingency table and the Chi

percentage of automatic normal results

Square

and

compared to manual reaching 94.4% and

examination result are shown in Table 1 and

high and low yields from both methods

2.

illustrate that the average the automatic

test.

The

with

results.

characteristic

Referring to WHO 2010 standard value

method shows that the normal results are

of normal spermatozoa morphology 4%. The

lower than the manual method shown as

conformity test used a 2 x 2 contingency table

83.3%.

from normal morphological data (Table 3).

Clinical Based Suitability

The results of this study could not be

According to the 2010 revision of WHO

analyzed. Because of the 36 samples tested

guidelines, men with normal sperm cell
59

Adhipireno Purwanto, et all.

morphology is ≥4%. This is observed by

94.4% and the high and low results of the two

namely by macroscopic and microscopic

methods

assessment of the semen (Table 4-5). The

automatic method results show more normal

sperm

results lower than the method. manual,

concentrationof

normal

results

automatically compared to manual, reaching

illustrating

that

the

average

namely 83.3%, as shown in Table 6.

Table 1. Subject Characteristics
Age (Year)
21 - 30
31- 40
> 40
amount

Ʃ
23
20
5
48

Sampling Technique
Masturbation
Coitus Interruptus
Special Condoms

Ʃ
48
0
0
48

Abstinence
(Day)
<2
2–7
>7

Ʃ
0
46
2

Table 2. Exclusion Research Samples
Description
Abstinence for more than 7 days
The sample is red / mixed with blood
Liquifaction more than 60 minutes
The concentration of spermatozoa is less than 2 million
Reagent Cassette damaged reading part is subject to hand grease
Amount

Ʃ
2
1
2
6
1
12

Statistical Based Conformity
Conformity Standard Mean between Automatic Normal Morphology and Manual Normal
Morphology
Table 3. Contingency 2x2 mean normal spermatozoa morphology
Automatic Normal Mean * Manual Normal Mean
Mean Normal Manual
Total
Positive
Negative
Automatic Normal Positive Count
10
1
11
Mean
% of Total
27.8%
2.8%
30.6%
Negative Count
7
18
25
% of Total
19.4%
50.0%
69.4%
Total
Count
17
19
36
% of Total
47.2%
52.8%
100.0%
Testing the results of research based on statistics using the standard mean value of the data;
Normal Morphology automatic method and Normal Morphology manual method.

-

Abnormal Head area automatic method and Abnormal Head area manual method

-

Abnormal Tail Length automatic method with Abnormal Tail length manual method.

Ina. J. Med. Lab. Sci. Tech. 2022; 4(1): 45–59

53

-

Adhipireno Purwanto, et all.

a.
b.
c.
d.

Sensitivity = 10/17 = 0.588%
Specificity = 18/19 = 0.947%
Positive prediction value = 10/11 = 0.909%
Negative prediction value = 18/25 = 0.720%

Table 4. Suitability test for the mean morphology of normal spermatozoa

Pearson Chi-Square
Continuity Correction
Likelihood Ratio
Fisher's Exact Test
Linear-by-Linear
Association
N of Valid Cases

Value

df

12,130 a
9,737
13,446

1
1
1

11,793

Asymp. Sig.
(2-sided)
0.000
0.002
0.000

1

Exact Sig.
(2-sided)

Exact Sig.
(1-sided)

0.001

0.001

0.001

36

The suitability test is based on the

Spermatozoa with slightly abnormal

calculation of the mean morphological value

'borderline' heads are classified as abnormal.

of spermatozoa with significance 0.001

The suitability test was based on the

<0.05. It can be concluded that H0 is rejected

calculation of the mean morphology of

and H1 is accepted, which means that by

abnormal

means of statistical analysis, there is a

significance of 0.956 > 0.05 (P < 0.05; r = -

difference between automatic spermatozoa

0.10). Then by means of statistical analysis,

morphology

we can observe that H0 is accepted and H1 is

and

manual

spermatozoa

morphology (Table 7).

spermatozoa

head

with

a

rejected, which means there is no difference
between

the

calculation

of

abnormal

Standard Conformance Mean Between

morphology of spermatozoa head automatic

Automatic Abnormal Head Area and

method with abnormal sperm morphology

Manual Abnormal Head

head manual methods.

Table 5. Contingency 2 x 2 mean abnormal head morphology
Mean Head Area * Mean Head Manual
Mean Head Manual
Mean
Head
Area
Total

Positive

Count
% of Total
Negative Count
% of Total

Positive
7
19.4%
8
22.2%
Count
% of Total

Negative
10
27.8%
11
30.6%
15
41.7%

Total
17
47.2%
19
52.8%
21
58.3%
59

Adhipireno Purwanto, et all.

a. Sensitivity = 7/15 = 0.467%
b. Specificity = 11/21 = 0.524%
c. Positive Predicted Value = 7/17 = 0.412%
d. Prediction Value Negative = 11 / 19 = 0,579%

Table 6. Suitability test for mean morphology of normal spermatozoa

Pearson Chi-Square
Continuity Correction
Likelihood Ratio
Fisher's Exact Test
Linear-by-Linear
Association
N of Valid Cases

Value

df

0.003
0.000
0.003

1
1
1

0.003

1

Asymp. Sig.
(2-sided)
0.955
1,000
0.955

Exact Sig
(2-sided)

Exact Sig.
(1-sided)

1,000

0.611

0.956

36

The suitability test was based on the

concluded that H0 is accepted and H1 is

calculation of the mean morphological value

rejected, which means that there is no

of abnormal spermatozoa tail with a

difference between the calculations of the

significance value of 0.339 > 0.05. Hence, by

abnormal spermatozoa tail morphology of the

means of statistical analysis, it can be

automatic method.

Standard Conformance Mean between Automatic Abnormal Tail and Manual Abnormal
Tail
Table 7. Contingency 2 x 2 mean abnormal morphology of tail
Automatic Tail Mean * Manual Tail Mean

Automatic Tail
Means

Positive

Count
% of Total
Negative Count
% of Total
Total
Count
% of Total
a. Sensitivity = 8/20 = 0.400%
b. Specificity = 7/16

Manual Mean Equipment
Positive
Negative
8
9
22.2%
25.0%
12
7
33.3%
19.4%
20
16
55.6%
44.4%

Total
17
47.2%
19
52.8%
36
100.0%

= 0.438%

c. Positive Prediction Value = 0.471%
d. Negative prediction value = 7/19 = 0.368%

55

Ina. J. Med. Lab. Sci. Tech. 2022; 4(1): 45–59

Adhipireno Purwanto, et all.

Table 8. Contingency 2 x 2 mean abnormal morphology of tail

Pearson Chi-Square
Continuity Correction
Likelihood Ratio
Fisher's Exact Test
Linear-by-Linear Association
N of Valid Cases

Value

df

0.942
0.403
0.945

1
1
1

Asymp. Sig.
(2-sided)
0.332
0.526
0.331

0.916
36

1

0.339

DISCUSSION

with

low

Exact Sig.
(2-sided)

Exact Sig.
(1-sided)

0.503

0.263

spermatozoa

concentrations.

This research was conducted at the

Because at concentrations below 2 million

Clinical Pathology Laboratory of Mother and

spermatozoa readings on the automatic

Child Hospital "Restu Ibu"

Sragen from

instrument will be read with a result <1 for

June 2020 to August 2020. Researchers carry

concentration, morphology and motility, and

out the rules of practice in the laboratory

then immediately remove the cassette from

examination stages as should the Pre-

the instrument after the reading is complete.

Analytic, Analytic and Post-Analytic.
Overall

in

this

study,

In

researchers

the

suitability

morphological

results

test
with

of

normal

automatic

conducted 48 patient samples, of which 12

methods compared to manual methods as

patient samples were included in the

Gold standard, using 2x2 contingency tables

exclusion criteria. 36 patient samples entered

to

the inclusion criteria according to the needs

predictive values and abnormal predictive

of the sample in this study. The sample

values. With the normal standards set by

characteristics of the 36 samples studied

WHO 2010, cannot be tested with 2x2

showed as many as 20.

contingency tables. Because the abnormal

see

sensitivity,

specificity,

normal

At the stage of carrying out the analysis

value in the manual method is 0. All results

using the Leenshoke SQA X1PRO automatic

from the 36 samples from the manual method

tool, there are several things that need to be

fall within the normal value range of ≥4

considered to get the results according to the

million/mL.

patient's clinical condition, namely: measure

Automatic morphological comparisons

the use of electricity used for automatic

that can be done clinically with WHO 2010

devices

sample

standards by looking at the number of normal

homogeneity, the cleanliness of the cassette

results achieved by the automatic method of

in the reading lens area must be completely

the 36 samples studied. The achievement was

clean, free of grease and dirt, cross checking

34 samples entered the normal range. This

reads using the manual method on samples

means that this automatic achievement

is

stable,

perfect

59

Adhipireno Purwanto, et all.

compared to the manual method reaches

c. Data on abnormal morphology of tails

94.4%. The normal morphological average

with a significance value of 0.339> 0.05.

yield of the automatic method is lower than

It can be concluded that H0 is accepted

the manual method. Data shows that of the 36

and H1 is rejected, which means that

samples, 83,3% lower than the manual

statistically

method.

between the calculation of abnormal

Because the standard normal value set by

there

is

no

difference

spermatozoa tail morphology automated

WHO 2010 cannot be clinically tested using

method

a 2 x 2 contingency table, the automatic

morphology manual methods.

method

The presence of an automatic tool

suitability

test

is

performed

with

abnormal

sperm

statistically by determining the standard of

LenshookeTM

the mean value. The statistical test consisted

advantages such as simple physical tools,

of data on normal morphology, abnormal

easy operation with touch screen technology,

morphology of the head and abnormal

quick reading of results that only takes 3 to 5

morphology of the tail. After testing the three

minutes, visualization of spermatozoa in the

data with the standard mean as standard, the

form of a video, focus setting and HDMI

following results are obtained:

connection to the monitor can display pH,

a. Normal morphological data shows a

motility, concentration and morphology.

significance value of 0.001 < 0.005, it can

Disadvantages of this device are in conditions

be concluded that H0 is rejected and H1 is

of low spermatozoa concentration, the device

accepted, which means that there is a

cannot read spermatozoa completely, and the

difference between statistical calculations

investment

for

with

operational costs for reagents are quite high

automatic method and manual method of

compared to manual ones. In the manual

spermatozoa morphology.

method with Papanicolaou dye, the results of

spermatozoa

morphology

SQA

costs

XI

for

PRO,

has

equipment

its

and

b. Data on abnormal morphology of the

the head, tail and cytoplasmic morphology of

head had a significance value of 0.371 >

spermatozoa were more clearly read than

0.05. It can be concluded that H0 is

other dyes. Tool investment costs and

accepted and H1 is rejected, which means

operating costs are lower than automated

that statistically there is no difference

methods. The data from the research were

between the calculation of abnormal

carried out by descriptive tests with the

morphology

results of the standard deviation of the

of

spermatozoa

head

morphology

morphology manual methods.

spermatozoa, the manual method showed the

Ina. J. Med. Lab. Sci. Tech. 2022; 4(1): 45–59

of

normal

or

abnormal

57

automatic method with abnormal sperm

Adhipireno Purwanto, et all.

results with the same value, namely 2.253

overall

and the standard deviation value of the

examination take into observation multiple

morphology

abnormal

aspects, namely a sperm’s shape such as its

spermatozoa, the automatic method also

head, midpiece, tail, and the presence of

showed the same results, namely 2.247.

cytoplasmic droplets. The most common

of

normal

/

In the suitability test for normal

fertility

potential.

Morphology

Spermatozoa morphology examination in

morphology results, the automatic method is

medical

laboratory

is

microscopic

compared to the manual method as the Gold

observation by manual

standard, using a 2x2 contingency table to see

machine. In general, the presence of the

the sensitivity, specificity, normal predictive

LenshookeTM SQA XIPRO device which is

value and abnormal predictive value, with the

based on automatic technology is compatible

normal standard set by WHO 2010, cannot be

with the manual method. Hence, this

tested with a 2x2 contingency table because

automated tool can be used in laboratory

the abnormal value in the manual method was

services complementing the manual method

0. All the results from 36 samples of the

as the Gold Standard.

and eutomatic

manual method were within the normal value
range of 4 million/mL. The automatic

AUTHOR CONTRIBUTIONS

morphological comparison that can be done

All author has equal contribution for the

clinically with the 2010 WHO standard is by

study conception and design, data collection,

looking at the number of normal results

analysis and interpretation of results, and

achieved by the automated method. Of the 36

manuscript preparation.

samples studied, the achievement is that as
many as 34 samples are in the normal range.
This means that if the automatic achievement
is compared to the manual method, it reaches
94.4%. The average yield of the normal
morphology of the automatic method is lower
than that of the manual method. The data
shows that of the 36 samples, 83.3% lower

ACKNOWLEDGEMENT
The authors would like to thank our
colleagues of at the Clinical Pathology
Laboratory of Mother and Child Hospital
"Restu Ibu” Sragen, their help is most
grateful. We also like to thank the patients
who participated in this study.

than the manual method.

CONCLUSION
Spermatozoa morphology examination is
important in terms of determining a man’s

CONFLICT OF INTEREST
The authors declare that there is no
conflict of interest.

59

Adhipireno Purwanto, et all.

REFERENCES

11. Bonraybio. Lenshooke XI PRO Semen quality
analyzer.
2019;
1-4.
http://bonraybio.com/en/product.php?act=view&
id=8.

1.

Lasiene K. Assessment of human sperm cells
morphological parameters. Spermatozoa - Facts
and Perspectives. 2018 ; 51-62.

2.

Susilowati T. Buku Spermatology. 1st Edition .
Malang. Universitas Brawijaya Press. 2011. 3-7p.

12. WHO. Laboratory manual for the examination
and rrocessing of human semen. 5 ed. WHO
Library. 2010.

3.

Tortora GJ, Derrickson B. Principles of anatomy
& physiology .14 ed. America . Kaye Pace . 2014.
1078p.

13. Auger J. Spermatozoa and sperm structure.
Second Edi. Vol. 1, Encyclopedia of
Reproduction. Elsevier; 2018. 62-67 p.

4.

Agarwal A, Henkel R, Huang CC, Lee MS.
Automation of human semen analysis using a
novel artificial intelligence optical microscopic
technology. Andrologia.2019; 2. 1-9.

5.

Chang V, Heutte L, Petitjean C, Härtel S,
Hitschfeld N. Automatic classification of human
sperm head morphology. Computers in Biology
and Medicine. 2017 ; 84.205-16.

14. Aksoy E, Aktan TM, Duman S, Cuce G.
Assessment of spermatozoa morphology under
Light Microscopy with different histologic stains
and comparison of morphometric measurements.
International Journal of Morphology. 2012 ; 30
(4):1544-550.

6.

7.

Danis RB, Samplaski MK. Sperm morphology:
History, challenges, and impact on natural and
assisted fertility. Current Urology Reports. 2019;
20. 43.
Rowe M, Albrecht T, Cramer ERA, Johnsen A,
Laskemoen T, Weir JT, et al. Postcopulatory
sexual selection is associated with accelerated
evolution of sperm morphology. Evolution.2015 ;
69 (4) :1044-52.

8.

Dearing C, Jayasena C, Lindsay K. Can the cperm
class analyser (sca) casa-Mot system for human
sperm motility analysis reduce imprecision and
operator subjectivity and improve semen
analysis?. Human Fertility . 2014; 17(1): 37–44.

9.

Lestari SW, Sari T. Fragmentasi DNA
spermatozoa penyebab deteksi dan implikasinya
pada infertilitas laki-laki Spermatozoa DNA
fragmentation detection and detection in male
infertility. E-Journal Kedokteran Indonesia.
2015;3(2):1–2.

10. Engel KM, Grunewald S, Schiller J, Paasch U.
Automated semen analysis by SQA Vision ®
versus the manual approach—A prospective
double-blind study. Andrologia. 2019 ; 51(1): 110.

15. Narulita P. The Effectiveness of LenshookeTM
Semen Quality Analyzer X1 Pro for human semen
analysis. Fac Med Univ Airlangga, Surabaya.
2020;1(1).
16. Sugiyono. Metode Penelitian Kuantitatif
Kualitatif dan R&D Quantitative Qualitative and
R&D.
Graha
Ilmu.
edisi1.Bandung,
Alfabeta.2019. 286 - 292p.
17. Amann RP, Waberski D. Computer-assisted
sperm analysis (casa): Capabilities and potential
developments. Theriogenology.2014 ; 81(1)5-17.
18. Apriora VD, Amir A, Khairsyaf O. Gambaran
morfologi spermatozoa pada perokok sedang di
lingkungan PE group yang datang ke Bagian
Biologi Fakultas Kedokteran Universitas Andalas
Description of Spermatozoa morphology in
medium smokers in the PE Group who came to
the Biology Section of the Faculty of Medicine,
Andalas University. J Kesehat Andalas.
2015;4(2):425–9.
19. Marzano G, Chiriacò MS, Primiceri E,
Dell’Aquila ME, Ramalho-Santos J, Zara V, et al.
Sperm selection in assisted reproduction: A
review of established methods and cutting-edge
possibilities. Biotechnology Advances. 2019 ; (
19 ) : 3.

59

Ina. J. Med. Lab. Sci. Tech. 2022; 4(1): 45–59