The semen analysis is the most important test for male fertility. It examines semen volume, sperm count, motility, morphology, and other factors. Abnormal results may indicate infertility issues. Hormone tests of blood and urine examine reproductive hormone levels which are important for ovulation. Ultrasound can check the ovaries, uterus, and fallopian tubes for issues like endometriosis that impact fertility. Other tests include hysterosalpingography which uses dye and x-rays to examine the fallopian tubes for blockages. Together these tests provide information to diagnose and treat fertility problems.

1. FERTILITY TESTS Lyndsae Drury

2. THE SEMEN ANALYSIS
The single most important test of male fertility
What are they looking for?
Sperm count
Ability of sperm to swim (motility)
Velocity or forward progression of the sperm
Size and shape of the sperm (morphology)
Total semen volume
The liquefaction of the semen (the ability to go from normal gel-like
state at ejaculation to a liquid state)

3. The Factors Reported in a Semen Analysis
A semen analysis is usually done at a laboratory or a physician's
office. The man masturbates and collects the ejaculate into a cup. The
semen should then be examined within a few hours, to achieve the
most accurate results. The following is evaluated:
Total Volume
2/5 milliliters is a normal volume. A very low volume indicates that
the seminal vesicles may not be making enough fluid or that these
ducts may be blocked. It may also indicate a problem with the
prostate gland.
Sperm Count
40 million to 300 million is the normal range for the number of
sperm per milliliter. Counts below 10 million are considered poor;
counts of 20 million or more may be fine if motility and morphology
are normal.

4. Liquefaction
Normal semen which is liquid at ejaculation immediately coagulates
into a pearly gel that liquefies within 20 minutes. Failure to coagulate
and then liquefy may indicate a problem with the seminal vesicles, as
would increased thickness or the presence of white blood cells.
Seminal Fructose
If no sperm are present, the semen will be tested for seminal
fructose, normally produced by the seminal vesicles. If no fructose is
present, congenital absence of the vas deferens or seminal vesicles or
obstruction of the ejaculatory duct
Cultures
The physician may test the semen and/or the urethra for the
presence of ay STD's or other bacteria.

5. MORPHOLOGY
AT LEAST 30% OF CELLS SHOULD
BE OF NORMAL SHAPE
ACCORDING TO THE WHO (WORLD
HEALTH ORGANIZATION)
KRUGER (STRICT)
MORPHOLOGY TEST
EXAMINES THE SHAPE AND SIZE OF THE
SPERM HEAD. NORMAL RESULTS ARE
WHEN 14% OR MORE OF THE SPERM
HAVE NORMAL SHAPED HEADS. MEN
WITH LESS THAN 4% OF NORMAL
SHAPED SPERM MAY HAVE A
SIGNIFICANT INFERTILITY PROBLEM.

7. Dark field microscopy is a
very simple yet effective
technique and well suited
for uses involving live and
unstained biological
samples. The Sperms
must be kept alive and
staining and heat fixing
would kill them .

8. BLOOD TESTS TO CHECK
REPRODUCTIVE HORMONESLevels of hormones in a woman's blood are closely linked to ovulation, when the
ovaries release an egg into the fallopian tubes. Hormone imbalances can cause
ovulation problems, and a blood test can help determine whether this is
happening. Going through a phase of not having periods, or having irregular
periods, are also signs of ovulation problems. The most common cause of
ovulation problems is polycystic ovary syndrome.
Progesterone is produced primarily by the corpus luteum of the ovary in
normally menstruating women and to a lesser extent by the adrenal cortex.
At approximately the 6th week of pregnancy, the placenta becomes the major
producer of progesterone.
2-5 The major functions of progesterone are in the preparation of the uterus for
implantation and maintenance of pregnancy, that is growing the endometrium.
Luteal phase concentration of Progesterone released by corpus luteum 10-
20ng/mL at 5 to 7days
Concentration of progesterone after conception, secreted by corpus luteum10-
50 ng/mL

9. Progesterone test is a carried out by the Chemiluminescent
Microparticle
Immunoassay (CMIA) for the quantitative determination of
progesterone in
human serum and plasma.
•Blood is collected by the Phlebotomist or the nurse.
•The sample is sent to the biomedical lab for analysis.
•The blood is labelled by the Biomedical technician and Bar
code married to patients NHS number on LIMS.
•Blood is centrifuged.
•Blood is mounted on the architect machine and the CMIA is then
carried out.
•Results are calculated by the architect machine and sent to the LIMS

10. Chemiluminesc
ent marker
(inactive until
trigger
chemical
added).

11. CHEMILUMINESCENCE FOR
FSH
Chemiluminescence is used to detect levels of FSH also. When Levels
of FSH are too low then the fertility expert can prescribe FSH tablets
to be taken to stimulate the development of follicles.
Revised LH test http://www.drg-international.com/ifu/rap-2753.pdf
accessed 30/11/2013

12. URINARY DIP STICKS
Urinary dip sticks for LH are also widely used for ovulation
detection, but are expensive, open to problems of
interpretation and are only of value when periods are regular.
Blood or urinary LH tests are of no value in general practice.
This LH Rapid Test is a qualitative, two site sandwich
immunoassay for the determination of human luteinizing
hormone in urine specimens.
The membrane was precoated with LH specific antibodies on
the test region.
During the test, the specimen is allowed to react with the LH
monoclonal antibody-colloid gold conjugate which was pre-
dried on the test strip.
The mixture then moves upward on the membrane
chromatographically by the capillary action.
For a positive specimen, the conjugate binds to the LH
forming an antibody-antigen complex.
This complex binds to the LH antibody as a capture regents
on the test region and produces a colored band when

13. 1. The urine specimen must be collected in a
clean dry container either plastic or glass,
without preservative. No centrifugation or
filtration of urine is required.
2. The test can be performed at anytime during
the day, however, for best results, the urine
sample for the test should be collected at about
the same time each day. Testing for the LH
peak should begin on the 10th day after the
beginning of a woman’s current menstrual
cycle. Testing should continue every day until
the LH peak is detected.
3. If specimens cannot be tested after
collection, they should be stored refrigerated at
2-8°C. If samples are refrigerated, they must be

14. As a general rule, swabs with wooden shafts should not be used.
Cotton, Dacron and calcium alginate swabs may all be used, although
toxicity has been noted with specific lots of each (6).
The cytobrush has also been used to collect endocervical specimens.
It appears to collect more cells than swabs and has been associated in
some investigators' experiences with higher recovery rates of
chlamydiae and higher rates of antigen detection by direct
fluorescent antibody (DFA)
Chlamydia is a bacteria, very short rods which may be
mistaken for cocci. They are Coccobacilli is Gram
negative.
Tests for Chlamydia

15. SPECULUM
CYTO BRUSH

17. The added
known probe
DNA

18. http://www.bi
ng.com/video
s/search?q=n
ucleic+acid+h
ybridisation&F
ORM=HDRSC3
#view=detail&
mid=EA613F0
3A556E1789C
02EA613F03A
556E1789C02

19. http://www.youtube.com/watch?v=jPV3LAp79Lc

20. Chlamydial inclusions were detected with C.
trachomatis-specific fluorescein isothiocyanate-
labeled monoclonal antibodies (green-yellow)
Commercial DFA testing kits are available, which
contain fluorescently labelled antibodies,
designed to specifically target unique antigens
present in the bacteria or virus, but not present
in mammals (Eukaryotes). This technique can be
used to quickly determine if a subject has a
specific viral or bacterial infection.EB =
ELEMENTARY
BODY
RB
=METABOLICALL
Y ACTIVE
RETICULATE
BODY (RB)

22. HOW TO STAIN A SLIDE
FLUORESCENTLY
https://courses.cit.cornell.edu/biomi290/microscopycases/methods/
FLASH%20DFA/The%20Direct%20Fluorescent%20Antibody%20Test.ht
m
First you must buy or prepare a fluorescein-labeled antibody. This
antibody must be specific for the organism or protein you are trying
to detect.
Prepare your sample by fixing it to the slide.
Add the fluorescein-labeled antibody. Give them time to bind,
Rinse off unbound antibody and observe the slide under a fluorescent
microscope. If the sample contains the antigen of interest, it will
emit light.

23. First barrier filter Lets
through only blue light with a
wavelength between 450nm
and 490nm
Second barrier filter cuts
out unwanted signals
passing the specific green
fluorescein emissions
between 520 and 560nm
Beam splitting mirror:
reflects light below 510nm
but transmits light above
510nm

24. ULTRASOUND SCAN
An ultrasound scan can be carried out to check the woman's ovaries,
womb and fallopian tubes. In a transvaginal ultrasound scan, which
takes place in hospital, a small ultrasound probe is placed in the
vagina.
Certain conditions, such as endometriosis and fibroids, can prevent
pregnancy from occurring.
The scan can also check for blockages in your fallopian tubes, which
may be stopping eggs from travelling along the tubes and into the
womb.
The Human Fertilisation & Embryology Authority (HFEA) estimates
that around one-third of women with fertility problems have blocked
or damaged fallopian tubes.

25. Ultrasound has been used by radiologists and sonographers to image
the human body for at least 50 years and has become a widely used
diagnostic tool. The technology is relatively inexpensive and portable,
especially when compared with other techniques, such as magnetic
resonance imaging (MRI) and computed tomography (CT)

26. PRINCIPLES OF SONAR

27. X-RAY OF FALLOPIAN TUBES
This is called a hysterosalpingogram (HSG). Opaque dye is injected
through the cervix while you have an X-ray. The dye will help your
doctors to see if there are any blockages in your fallopian tubes.
Blockages can prevent eggs passing down the tubes to the womb,
and so stop pregnancy occurring. Find out more about testing for and
diagnosing infertility problems.
• The woman is positioned under a
fluoroscope (a real-time x-ray imager)
on a table.
• The gynaecologist or radiologist then
examines her uterus and places a
speculum in her vagina.
• Her cervix is cleaned, and a device
(cannula) is placed into the opening of
the cervix.
• The doctor then gently fills the uterus
with a liquid containing iodine (contrast)
through the cannula.
• The contrast then enters the tubes,
outlines the length of the tubes, and
spills out their ends if they are open.

28. HYSTEROSALPINGOGRAM

29. REFERENCES
https://courses.cit.cornell.edu/biomi290/microscopycases/methods/f
abs.htm
Direct Fluorescent Antibody Test

30. Yingxue Zhang, Zhujun Zhang, Feng Yang, 0ctober 2007, Analysis of
Follicle-Stimulating Hormone by CE with Chemiluminescence
Detection Based on Competitive Immunoassay, Chromatographia,
Volume 66, Issue 7-8, pp 539-544
NHS Fertility tests. http://www.nhs.uk/conditions/pregnancy-and-
baby/pages/fertility-tests.aspx Accessed 30/11/2013.
Robert J. Norman, Australian Prescriber, Fertility tests
http://www.australianprescriber.com/magazine/25/2/38/40/.
Accessed 30/11/2013
Screening tests to detect chlamydia
http://www.cdc.gov/mmwr/pdf/rr/rr5115.pdf
2002, accessed 30.11.2013

31. Gary England, Michelle Kutzler, Pierre Comizzoli, Wojciech Nizanski, Tom
Rijsselaere And Patrick Concannon. ISCFR 2012 July Congress of the European
Veterinary Society for Small Animal Reproduction
http://www.ivis.org/proceedings/iscfr/2012/108.pdf?LA=1 accessed
30/11/2013
Progesterone, Architect manual, revised April 2010
http://www.ilexmedical.com/files/PDF/Progesterone_ARC.pdf accessed
30/11/2013
Robert C. Brunham & José Rey-Ladino, (February 2005), The development of
Cylamydia Trachomatis
http://www.nature.com/nri/journal/v5/n2/fig_tab/nri1551_F2.html Nature
Reviews Immunology 5, 149-161 Accessed 30/11/2013
Max A Chernesky, PhD, Can J Infect Dis Med Microbiol. 2005 Jan-Feb; 16(1): 39–