SEMEN

-secretion of the male reproductive system composed of fractions contributed by the testes, epididymis, seminal vesicles,
prostate and the cowper’s gland (Bulbourethral gland).

Origin and Composition

Testes Site of sperm production

Seminiferous tubules Found in the testes, which is

responsible for forming the
sperm.

Spermatozoa are produced in

the epithelial cells of these
structures

Sertoli Cells “Nurse Cells” Provides nutrients and support

to the germ cell cells as they
undergo meiosis and mitosis
(SPERMATOGENESIS)

Epididymis Maturation of non-motile

immature sperm after
spermatogenesis.

Spermatozoa develops flagella.

Spermatozoa remains here until

ejaculation.

SPERMATOZOA→ 1° SPERMATOCYE→ 2° SPERMATOCYTE→ SPERMATIDE (W/O TAIL)→ EPIDIDYMIS MATURATION→

SPERMATOCYTE

Composition of Semen
Percentage Main Component

Spermatozoa 5% Sperm cells

Seminal Fluid 60-70% Fructose

Prostate Fluid 20-30% ACP, Zinc, Citric acid,

Ascorbic acid,
enzymes

Bulbourethral 5% Alkaline Mucus

secretions

Specimen Collection
-Abstinence:

 2-3 days but not more than 5 days, before the test.
  Excessive abstinence- ↑volume and ↓motility

-Should be delivered fresh, not longer than 1 hour after collection.

-Fertility Testing:

 2-3 samples collected not less than 7 days or more than 3 weeks apart.

 2 abnormal samples is considered significant

- Masturbation- most preferred method of collection.

Other methods: Emission after coltus interrupts,Condom collection, silastic condom collection, vaginal vault
aspiration, epididymal extraction.

- If not possible- ordinary, non-lubricant containing rubber or polyurethane should be used.

-Collection must be complete because the first portion contains the concentrate

-Specimen should be kept at 37°C prior to testing (Seminal Banks- -85°C for 1 year)

Semen Analysis
Appearance

Appearance Description

Gray-white color, transluscent Normal

and has a musty odor

Red Presence of Blood

Yellow Urine contamination

Specimen collection following

prolonged abstinence and
medication

Clear Infertility

Increased white turbidity WBCs

Liquefaction
- should liquefy within 30-60 minutes minutes after collection.

-If after 2 hours the specimen failed to liquefy- add Physiologic Dulbecco’s Phosphate Buffer saline or alpha-chymotrypsin or
bromelain

Reference Values for Semen Analysis

Volume 2-5 mL

Viscosity Pours in droplets

 pH 7.2-8.0

Sperm Concentration >20million/mL

Sperm Count >40 million/ejaculate

Motility >50% within 1 hour

Quality >2.0 or a,b,c in Sperm motility

grading

Morphology >14% normal forms (strict

criteria)

>30% normal forms (routine

criteria)

Round cells <1.0 million/mL

*Concentration- number of sperm cells per mL of the specimen.

Sperm Concentration is counted using Neubauer counting chamber.

Formula:

# of cells counted x DF -# of squares counted must be expressed to large squares

Sperm Concentrat ion 
# of squares counted x depth
(1 large square= 25 small squares) and sperms are counted in 5 small square (5/25=0.2)

-dilution can be achieved using a positive-displacement pipet. 1:20 is the most common DF.

Diluents used:

1. Sodium bicarbonate with formalin (immobilizes the sperm)

2. Saline and cold distilled water

*Results are always multiplied by 1,000 because the unit for sperm conentration must be expressed in mL

Unit: cells/mL

*Count- number of sperm cells in the whole volume of the specimen given.

Formula:

Sperm count = Sperm Concentration x Total Volume

Unit: cells/ejaculate

Sperm Motility Grading

Traditional Scale WHO Grading WHO Criteria

4.0 a Rapid, Straight line

motility

3.0 b Slower speed, some

lateral movement
 2.0 b Slow forward
progression,
noticeable lateral
movement

1.0 c No forward
progression

0 d No movement

*Within 1 hour, 50% or more sperm should be motile in categories a,b and c or 25% or more should show progressive motility
in a and b.

*Computer-Assisted Semen Analysis (CASA)- new automated sysem used to assess sperm motility. It is able to detect sperm
velocity and trajectory.

Alternative Sperm Motility Grading Criteria (WHO, 2010)

Progressive Motility (PM) Sperm moving linearly in a large

circle

Nonprogressive Motility (NP) Sperm moving with an absence

of progression

Immotility (IM) No movement

Sperm Morphology
 Must evaluate 200 sperm cells

Routine criteria: >30% normal

Kruger’s strict criteria: >14% normal

 Normal sperm specifications

Head Oval in shape; 5µm long and 3µm

wide

Tail (Flagella) 45µm long

Midpiece Connects the head and the tail

7µm long and the thickest part of

the tail because of mitochondrial
sheath that produces energy
required for motility

Acrosomal Cap Encompasses half of the head

and cover approximately 2/3 of
sperm nucleus

*To assess mosrphology, use Giemsa, Wright’s, Shorr or Papanicolau as the stain.
*Varicocoele- most common cause of male infertility. (Tapered Head).

Sperm Vaibility/ Vitality/ Bloom’s Test

 Determine if the individual has live or dead sperms

 Eosin-nigrosin- main stain used

 100 cells are counted

 Live sperm cells- bluish-white

 Dead sperm cells- red/ orange

 Normal viability should have atleast 75% living cells

Seminal Fluid Fructose

 Used to asses function of the seminal vesicles

 Resorcinol-HCL test is used

 Positive result: orange

 Normal Values: equal to or >13umol per ejaculate

Antisperm Antibodies
 Can be present in both males and females

 Two tests available:

 Mixed agglutination Reaction (MAR)- screening procedure to detect IgG Abs against sperm cells

 Immunobead test- more specific since t can detect IgG, IgM and IgA antibodies against the specific area of perm cell.

Other Tests for Seminal Fluid

Florence Test Test for choline; uses potassium
iodide and iodine

Barbiero’s Test Test for Spermine; uses Picric

acid and TCA

Spinnbarkeit’s Test Test for tenacity of Mucus

Hamster’s Egg Penetration Test for penetration of sperm;

uses hamster egg cells