1. Semen analysis

2. Part I Semen collection Fertility has nothing to do with potency Does size matter? Concerns about the size of genital are often unwarranted. Other researcher say it does.

3. The penis of Rasputin measuring 28
The penis of Rasputin measuring 28.5 cm (about 11 inches) in st Petersburg Museum example of gender obsession gone wrong ending terribly for owner of this member.

4. Why birds do not have penises
Why birds do not have penises ? Because of a genetic signal causes the penis cells to die off during gestation.

5. Semen collection By sexual act

7. Collection of semen in a condom

8. Assaf Harofeh Medical Center's sperm bank
Assaf Harofeh Medical Center's sperm bank. A court has allowed the parents of a dead man to chose a woman to be impregnated by his sperm, which is stored in a similar facility. Photo by Moti Milrod

9. Semen cryopreservation

10. ARTIFICIAL INSEMINATION

11. Intrafallopian Insemination (IFI)
The intrafallopian insemination is ideal for the following conditions:
@ Women with ovulation issues
@ Women with cervical mucus that prevents pregnancy
@ Used as a last chance method for couples when other types of artificial insemination have failed

12. Microsurgical Epididymal Sperm Aspiration
Next, the dilated epididymal tubules are opened with an opthalmological blade.  The epididymal fluid is aspirated into a syringe using an angiocath tip.  The fluid is examined for the presence of motile sperm and then sent to the andrology laboratory for cryopreservation
Microsurgical Epididymal Sperm Aspiration

13. Microdissection Testicular Sperm Extraction
The tunica albuginea is opened transversely along the equator of the testicle.  Use of the operating microscope allows for excellent visualization of vital structures and lowers the risk of any damage to the testicle.  Once the testicle is opened, a large surface area of tissue can then be safely inspected for areas of active spermatogenesis.  The microsurgeon holds the testicle while the assistant uses irrigation  for optimal visualization.   
Microdissection Testicular Sperm Extraction

14. The psychobiology of human semen

15. Timing sex
If you’re trying to get pregnant, something useful to know is how often to have sex.  Sperm counts peak at two to three days after ejaculation, meaning, after you have sex (or masturbate,) the swimmers will be most abundant two to three days later.  Sperm live for about 48 hours in the female reproductive tract, so the optimum timing is sex every other day.
I’m often asked when during the month (or cycle) a couple should try to conceive.  Studies show that conception usually happens up to six days before, and ending on, ovulation.  The big problem is that all tests for ovulation show the event after it happens.  So a good plan is start having sex every other day after the beginning of menstruation by 9 days, and keep going every other day for 5 times ( the possible days of ovulation) .Then be at your ease until the next menstruation

16. Part II Semen
Sperm count, or sperm concentration to avoid confusion with total sperm count, measures the concentration of sperm in a man's ejaculate, distinguished from total sperm count, which is the sperm count multiplied with volume. Over 15 million sperm per milliliter is considered normal, according to the WHO in Older definitions state 20 million. A lower sperm count is considered oligozoospermia.
Motility
The World Health Organization has a value of 50% and this must be measured within 60 minutes of collection. WHO also has a parameter of vitality, with a lower reference limit of 60% live spermatozoa.
Morphology
Regarding sperm morphology, the WHO criteria as described in 2010 state that a sample is normal (samples from men whose partners had a pregnancy in the last 12 months) if 4% (or 5th centile) or more of the observed sperm have normal morphology.
Morphology is a predictor of success in fertilizing oocytes during in vitro fertilization.
Up to 10% of all spermatozoa have observable defects and as such are disadvantaged in terms of fertilising an oocyte.
Also, sperm cells with tail-tip swelling patterns generally have lower frequency of aneuploidy.
Volume
Web MD advises that semen volumes between 1.0 mL and 6.5 mL are normal; WHO regards 1.5 ml as the lower reference limit. Low volume may indicate partial or complete blockage of the seminal vesicles, or that the man was born without seminal vesicle.
Fructose level
Regarding the level of fructose in the semen, Web MD lists normal as at least 3 mg/mL. WHO specifies a normal level of 13 μmol per sample. Absence of fructose may indicate a problem with the seminal vesicles. pH
Web MD lists a normal pH range of ; WHO criteria specify normal as Acidic ejaculate (lower pH value) may indicate one or both of the seminal vesicles are blocked. A basic ejaculate (higher pH value) may indicate an infection.
Liquefaction
The liquefaction is the process when the gel formed by proteins from the seminal vesicles is broken up and the semen becomes more liquid. It normally takes less than 20 minutes for the sample to change from a thick gel into a liquid.
MOT
MOT is a measure of how many million sperm cells per ml are highly motile, that is, approximately of grade a (>25 micrometer per 5 sek. at room temperature) and grade b (>25 micrometer per 25 sek. at room temperature). Thus, it is a combination of sperm count and motility.
spermatozoa
Total motile spermatozoa (TMS) or total motile sperm count (TMSC) is a combination of sperm count, motility and volume, measuring how many million sperm cells in an entire ejaculate are motile..
Others
The sample may also be tested for white blood cells. A high level of white blood cells in semen is called leucospermia and may indicate an infection.[1] Cutoffs may vary, but an example cutoff is over 1 million white blood cells per milliliter of semen
Measurement methods
Computer Assisted Semen Analysis (CASA) is a catch-all phrase for automatic or semi-automatic semen analysis techniques. Most systems are based on image analysis, but alternative methods exist such as tracking cell movement on a digitizing tablet. Computer-assisted techniques are most-often used for the assessment of sperm concentration and mobility characteristics, such as velocity and linear velocity. Nowadays, there are CASA systems, based on image analysis and using new techniques, with near perfect results, and doing full analysis in a few seconds. With some techniques, sperm concentration and motility measurements are at least as reliable as current manual methods.
Raman spectroscopy has made progress in its ability to perform the characterization 7 the perform of DNA damage.

17. WHO 4th semen reference vazlues

20. Kruger’s strict criteria for sperm morphology
The scientific validity of Kruger’s grading scheme is based on the fact that what normal sperm looked like was based on looking at sperm that had proven themselves as functionally competent, instead of looking at population of men whose partners became pregnant in less than a year.
. Kruger’s definition of normal was based on examining sperm that were able to successfully migrate through the cervical mucous on their way to the egg rather than looking for shared traits among a group of presumed fertile men. .. 
Kruger Strict analysis requires that each apparently normal sperm be measured for head size.  According to Kruger, the head length for normal spermatozoa may vary between 4.0 and 4.5 μm, with a mean length of 4.07 ± 0.19 μm and a mean width of 2.98 ± 0.14 μm. This precision measurement requires  an eye-piece micrometer to be placed in the microscope tube, projecting a “ruler” onto the image of the sperm so that the sperm head can actually be measured.
What is a normal sperm by Kruger’s strict criteria? A normal sperm head must be oval with a well-defined acrosome that covers 40-70% of the head.  The normal head has a length of µm and a width of 3 µm with very little allowance in either direction (plus or minus 0.14 µm).  The length to width ratio should be about 1.5. All borderline-normal head forms and/or spermatozoa with nearly oval heads with no gross abnormalities are still classified as abnormal if they don’t meet the head size criteria. Any abnormalities in the neck, midpiece or tail also automatically render the sperm abnormal regardless of head quality. The normal midpiece length is 1.5 times the head length, the width of the midpiece is less than 1 µm and is slender and attached to the tail. The tail is 45 µm in length, is not coiled and is slightly thinner than the midpiece. In sperm production, left over cytoplasmic drops are produced and may persist but are only  considered normal if their size does not exceed ½ the size of the sperm head.
Is it any wonder that most sperm techs hate this tedious test?
What is the good news for patients with poor morphology results? Regardless of your semen analysis result, as long as you have some recoverable sperm with normal DNA and signs of life (even a twitch will do) that will fit into the pipette,  a good ICSI tech can push that sperm into your partner’s eggs and you can still have children. So don’t despair over abnormal morphology results. Very low numbers of normal forms may suggest that IVF or ICSI may be necessary, but technical treatments are possible to overcome even very poor morphology results.

21. Human genital critical periods

22. Semen stain on carpet observed with and without ultraviolet light

23. Structural diagram of spermatozoon
A. Head:
a. Nucleus: contain genetic code b. Postnuclear cap: cover posterior portion of nucleus c. Acrosome: cover anterior portion of nucleus and contain enzymes as hyaluronidase,corona penetrating enzyme and acrosin
B. Neck:(sometimes included in "Tail") Proximal centriole: join head and tail, where head and tail separate during fertilization and in heat-damaged semen
C. Tail:
a. Mid-piece: Mitochondrial sheath: contain enzymes converting energy substrates as fructose into ATP b. Main-piece: Axial filament of 2 central fibrils, 9 pairs of inner fibrils and 9 coarse outer fibrils: its contraction by energy from ATP¡æ tail movement c. End-piece:

24. Normal anatomy of spermatozoa

25. Seminal plasma Seminal plasma
A. Function: Serve as buffer, optimal osmotic and nutrient medium
B. pH: nearly 7.0; slightly acidic in bulls and rams; slightly alkaline in boars and stallions
C. Osmotic pressure: similar to blood and physiological saline(0.9% NaCl)
D. Energy substrates: fructose, sorbitol etc
Inorganic ions
A. Major ions: Na+, Cl-, K+; minor ions : Ca++, Mg++
B. K+/Na+ ratio: High in sperm cell and low in seminal plasma
C. Function: Maintain optimal osmotic pressure for sperm survival
Buffering agents
A. Principal organic ion as buffering agent: Bicarbonate(HCO3-)
B. Source: Vesicular gland
C. Function: guard against change in pH of semen, but not sufficient
Energy substrates
A. Energy substrates: Fructose, sorbitol, glycerylphosphocholine(GPC)
B. Source: Fructose and sorbitol: vesicular gland; GPC: epididymis: uniquely high in semen
C. Metabolism:
a. Fructose: used under anaerobic and aerobic conditions b. Sorbitol and GPC:used only aerobically c. GPC: utilized after splitted into choline and glycerylphosphate by an enzyme in female tract
Other organic compounds
A. Inositol and citric acid: considerably high, but not utilized.
B. Ergothionine: found in boar and stallion semen
Energy metabolism by spermatozoa
*. Plasmalogen: a lipid reserved within sperm cell: used when other substrates are limiting
A. Anaerobically:Fructose ¢¢ 2 lactic acid + 2 ATP (net yield)
B. Aerobically : Fructose ¢¢ CO2 + H2O + 38 ATP (net yield)
C. ATP + H2O ¢¢ ADP + H3PO4 + 7,000 calories /mole

26. Seminal plasma ( continue)
Factors affecting rate of metabolism
A. Measurement of metabolism rate
A .Under aerobic condition: 1). O2 consumption 2). CO2 liberation 3) Methylene blue reduction time b. Under anaerobic condition: 1) pH reduction 2) Increase in lactic acid 3) Decrease in fructose
B. Relationship of fertile life to metabolic rate of sperm:
a. Reduced metabolic rate : extend the storage life of semen; b. Reduced metabolic rate in epididymis: extend life of epididymal sperm c. Sperm in fresh ejaculate of semen: fertile for a few hours under high rate of metabolism
Temperature
A. High temperature: Increase metabolic rate and decrease life span of sperm
B. At 50¡É: Irreversible loss of sperm motility
C. At body temperature: Sperm live for a few hours only: due to increase metabolism
D. Low temperature: Extend fertile life of sperm by decrease metabolism ( when bull semen frozen at -196¡É, less than 0.02% of metabolic rate at body temperature): Problems: cold shock and freeze kill
a. Cold shock: 1) Irreversible loss of sperm motility by sudden reduction of semen temperature from 15¡É to 0¡É (critical range), 2) Protecting from cold shock: Slow cooling after addition of lecithin and lipoproteins by diluting with egg yolk or milk diluter b. Freeze kill: 1) Sperm killed during freezing and thawing 2) Protected satisfactorily by equilibrating bull semen in a diluter containing glycerol ¡æ Desirable fertility from semen frozen for decades pH
A. Higher metabolic rate: From pH of semen near neutrality(7.0), where most enzymes in sperm are most active.
B. Deviations toward alkalinity or acidity: Can reduce metabolic rate, but buffering capacity of diluter is rather important because pH range to be altered without permanent impairment of sperm is narrow
Osmotic pressure
A. Semen diluted with isotonic diluter maintains maximum metabolic activity
B. Hypo- or hypertonic diluter: Can reduce metabolism but detrimental to sperm
Concentration of spermatozoa
A. Increase concentration of sperm ¡æ Inc. K+/Na+ ratio ¡æinhibit sperm metabolism
B. Moderate dilution with buffered, isotonic medium containing fructose, before cooling: Not greatly alter metabolic rate, but extend sperm life
C. Excessive dilution (over 1,000x): Depress motility and metabolic rate

27. Seminal plasma ( continue)
Gases
A. CO2
a. 5-10% partial pressure of CO2: ¡é sperm metabolism b. Epididymal sperm: Long life by high CO2 c. CO2 gassing in diluter: Effective for sperm preservation at room temperature
B. O2
a. Necessary for aerobic metabolism b. High level of O2: Toxic C. Other gases: N2, H2, He: No effects
Light
A. Light under oxygen: Produce harmful H2O2; Semen should be protected from light
B. H2O2 production: Prevented by addition of catalase to diluter
Antibacterial agents
A. Penicillin and dihydrostreptomycin or neomycin: Used for control of bacterial growth, with no effects on sperm metabolism
B. Antibacterial agents: ¡.e fertility from bulls infected with fibriosis
C. Control of bacteria in semen by antibiotics ¡æ Sparing energy substrates: Extend fertile life of sperm

28. Immature germ cells The number of immature germ cells should be ≤106/ ml

29. Spermin crystal Crystals of spermine phosphate were first described in 1678, in human semen, by Anton van Leeuwenhoek

30. Semen is a team sport
This old study demonstrates that women whose husbands have low sperm counts do not get pregnant as often as women whose husband’s have NO sperm. Consider the previous concept that some men with low counts have highly fertile partners. Those couples make babies without our help. Only the couples where the man and the woman are both subfertile would come for donor sperm insemination.
The idea of infertility treatment being a team sport is a useful concept to understanding that fertility disorders must always consider both parties.
Strict sperm morphology affects fertilization at IVF. The role of strict sperm morphology became evident in the 1980s as illustrated in the following graphic

31. Notice in this example that the man in each pair has a score of three
Notice in this example that the man in each pair has a score of three. If his partner is also a 3 then they have infertility. If the partner’s score is 5 then they have normal fertility (combined score = 8).

32. Elephant phallus preserved

33. Semen collection by manual collection technique

34. Part III Semen abnormalities
Nomenclature related to semen quality  
Aspermia - no semen (no or retrograde ejaculation).  
Asthenozoospermia - percentage of progressively motile spermatozoa (A+B classes) below the lower reference limit.  
Asthenoteratozoospermia - percentages of both progressively motile (A+B classes) and morphologically normal spermatozoa below the lower reference limits.  
Azoospermia - no spermatozoa in the ejaculate. 
 Cryptozoospermia - spermatozoa absent from fresh preparations but observed in a centrifuged pellet.  
Haemospermia (haematospermia) - presence of erythrocytes in the ejaculate.  
Leukospermia (leukocytospermia, pyospermia) - presence of leukocytes in the ejaculate above the threshold value.  
Necrozoospermia - low percentage of live, and high percentage of immotile, spermatozoa in the ejaculate.  
Normozoospermia - total number or concentration of spermatozoa, and percentages of progressively motile (A+B classes) and morphologically normal spermatozoa, equal to or above the lower reference limits.  
Oligoasthenozoospermia - total number or concentration of spermatozoa, and percentages of progressively motile (A+B classes), below the lower reference limits. 
 Oligoasthenoteratozoospermia - total number or concentration of spermatozoa, and percentages of both progressively motile (A+B classes)and morphologically normal spermatozoa, below the lower reference limits.  
Oligoteratozoospermia - total number or concentration of spermatozoa, and percentage of morphologically normal spermatozoa, below the lower reference limits. 
 Oligozoospermia - total number or concentration of spermatozoa below the lower reference limit.  
Teratozoospermia - percentage of morphologically normal spermatozoa below the lower reference limit.   
Main soft ware features
Automated analysis of sperm concentration and motility on native samples according to WHO recommendations.
Automated morphology analysis on stained samples according to strict Krueger’s criteria.
Manual assessment of concentration of white blood cells, immature germ cells, round cells.
Manual measurements for special research or other individual tasks.

35. Hormonal evaluation ENDOCRINE PROFILE ON INFERTILE MEN
STATUS Testosterone FSH LH PRL
Normal male Normal Normal Normal Normal
Primary testicular failure Low HighNormal/ High Normal
Hypogonadootrophic hypogonadism
Low Low Low Normal
Hyperprolactinemia Low Low/ Normal LowHigh
Androgen resistance High High High Normal
Hormonal Evaluation: Evaluation of Brain - Testicle (Hypophyso- gonadal) axis provides information about sperm production. FSH and testosterone level should be tested for pathologies occurred over hypophyseal axis (hyperprolactinemia, gonadotrophine deficiency, congenital adrenal hyperplasia) and patients who have sperm count below 10 million. Testosterone is the end product of hypophyseal axis and provides information about general endocrine balance of the reproductive system. If endocrine abnormality is present, serum LH and prolactin levels may be tested to detect the location of the pathology. Estradiol, Thyroid hormones, Liver tests may be ordered. Decrease on LH and FSH in normal spermatogenesis is not significant. Also, low LH and normal testosterone are not significant. In brief; conditions that require hormonal assessment: 1) If sperm density is below 10 million /ml 2) If there is disrupted sexual function 3) If there are specific endocrinopathy findings (hormonal diseases)

36. Morphological abnormalities of spermatozoa
Abnormal morphology
A. Reduced fertility: From 25% or more abnormal sperm
B. Classification of abnormal sperm
Morphological abnormalities of spermatozoa identified through examination of semen for quality. 
a. Abnormal heads: asymmetrical, tapering , pyriform, giant, micro and double heads b. Abnormal tails: enlarged, broken, bent, filiform, truncated and double mid-pieces; coiled, looped and double tails; cytoplasmic droplets (formed on neck during spermiogenesis and usually lost during maturation)

37. Morphological defects in sperm
Free head
Round head
Double , triple head
Bent neck

38. Morphological defects in sperm
Dag defect
Coiled tail
Double tail
Tail stump

39. Round cells in semen analysis
Round cells are often encountered during semen analysis. It is difficult to distinguish immature sperm cells (heads without tails) from white blood cells without specialized stains.
Sperm cells that have not completed maturation may not have a tail. These cells can look alot like white blood cells (WBCs). WBCs are normal components of blood. An elevated concentration of WBCs in a semen analysis suggests the presence of an infection (often prostatitis).
Methods to detect WBCs (by distinguishing these from immature sperm cells) vary. A simple peroxidase stain is able to identify (nonactivated) WBCs since these WBCs contain abundant peroxidase enzyme. WBCs also have a specific immunoreactive region (called the CD 45 antigen) that can be identified with specific (monoclonal) antibodies that attach to this antigenic site.

40. Hematosperemia
The following conditions have been reported in association with hematospermia:
Benign or malignant tumors of the prostate, bladder, testes, or seminal vesicles
Infections including, but not limited to, chlamydia , herpes , cytomegalovirus and trichomoniasis.
Inflammation of the prostate (prostatitis), epididymis (epididymitis), or urethra (urethritis)
Calculi (stones similar to kidney stone ) in the seminal vesicles or prostate
Polyps in the urethra
Ejaculation duct obstructions
Metastatic cancers (that have spread from other sites in the body) located in the genitourinary system
Cysts, hemorrhage, or other abnormalities in the seminal vesicles

41. Ultrastructural and mutational analysis of human PCD cases with axonemal disorganization in Kartagener syndrome.
Primary ciliary dyskinesia (PCD) is an inherited disorder characterized by recurrent infections of the upper and lower respiratory tract, reduced fertility in males and situs inversus in about 50% of affected individuals (Kartagener syndrome). It is caused by motility defects in the respiratory cilia that are responsible for airway clearance, the flagella that propel sperm cells and the nodal monocilia that determine left-right asymmetry. Recessive mutations that cause PCD have been identified in genes encoding components of the outer dynein arms, radial spokes and cytoplasmic pre-assembly factors of axonemal dyneins, but these mutations account for only about 50% of cases of PCD.

43. How do the seminal vesicles make semen
How do the seminal vesicles make semen? Do they produce it immediately or do they store it for later?
Only 50-70% of semen comes from the seminal vesicles (SVs). The fluid contains fructose for sperm to use for energy among other supporting elements. The SVs continuously make semen 24/7. If you were to record your "output" over a day it would likely be close to 8-10 cc/day. Since an average ejaculate is about 3.5 cc, you would see reduction for any additional samples obtained in the same day.
Seminal plasma proteins arise from secretions from the seminal vesicles (~65% of semen volume), prostate (~25%), testes and epididymides (~10%) and bulbourethral and periurethral glands (~1%).

44. Semen fructose
Fructose is a sugar produced by the seminal vesicles and provides energy for sperm motility. Its absence suggests a block in the male reproductive tract in ejaculatory duct. In men with no sperm or very low numbers of sperm in the ejaculate, it is important to determine whether the sperm are not being produced at all, or whether they are being produced but are blocked from getting into the semen. A fructose test can help differentiate between these two problems

46. Cell Phone Radiation Damages Male Fertility

47. The world only penis museum

48. A museum showing animal genitalia of all shapes & sizes – from whales to field mice museum. Over 2OO specimens are preserved while others are displayed the hunting trophies on the wall

49. Part IV Semen analysis
A. The quality of manual microscopic semen analysis varies enormously from centre to centre and despite efforts in more recent times to introduce standardization and quality control, it remains a time consuming and poorly reproducible exercise. .
B, Automated methods have been around for a number of years and have been proposed as both a time saving solution and one which will improve reproducibility but are still not widely adopted. There are several commercial systems as semen analyzer. Inconsistencies in existing and earlier CASA systems appear to be due to a number of factors which would include both the lack of clear validation and to a degree the technical expertise of the CASA user. 

50. A. Manual Semen analysis

52. Sperm count vary over time
Sperm counts vary over time– Sperm counts and motility can vary a great deal from one week to the next. The chart at right shows sperm counts for one man taken every two weeks over two years. As you can see his sperm counts varied widely
WHO laboratory manual for the examination of human semen and sperm-cervical mucus interaction, Fourth Edition

54. B. Automated analysis Sperm Class Analyzer® (SCA®) is a system for the quantitative analysis and management of the most important human semen parameters according to W.H.O. critera

55. Sperm Class Analyzer® (SCA®) is a system for the quantitative analysis and management of the most important human semen parameters according to W.H.O. critera
Motility
Morphology
Vitality
Fragmentation

56. SCA® Motility and concentration - automatic analysis of semen parameters

57. SCA® Motility and concentration - automatic analysis of semen parameters
The software detects the motile/immotile spermatozoa automatically, performs an accurate count and accordingly provides the concentration results. At the same time, motile sperm analyses is performed by following the sperm trajectory and determines the velocity. The results are then presented within seconds taking into account the W.H.O. motility parameters (both percentage motility ratings as well as the a to d WHO ratings): VCL, VAP, VSL, STR, LIN, WOB, ALH, BCF.
According to W.H.O. analysis of 200 sperm is required, which is approximately equivalent to 2 or 3 fields to be captured. Analysis time is one second per field.
SCA® has Full and Lite versions for motility and concentration

58. SCA® Morphology for automatic analysis of morphometry and morphology of sperm
According to the quantitative analysis of morphometry and morphology, the sperm is classified as normal or abnormal. There are several accepted classification criteria: W.H.O and strict Tygerberg, also known as Kruger, are both included in the SCA®. SCA® Morphology has the following editions: SCA® Morphology FULL, and SCA® Morphology LITE. Besides these classifications, SCA® Morphology FULL contains the User Defined criteria, which convert it in the ideal tool for research and development of new classification methods. SCA® MORPHOLOGY LITE contains only the W.H.O. criteria. It is possible to upgrade from lite to full edition.

59. SCA® DNA Fragmentation for the automatic analysis of DNA fragmentation in semen
SCA® DNA Fragmentation detects the spermatozoa in the captured images and classify them into Fragmented and non Fragmented. SCA® DNA Fragmentation is the only software recommended by the Autonomus University of Madrid (U.A.M.)

60. SCA® DNA Fragmentation for the automatic analysis of DNA fragmentation in semen

61. SCA® VITALITY for the automatic analysis of sperm viability
The analysis is done using fluorescence microscopy, in sperm samples treated with fluorochromes like Duo-Vital. Automatic recognition and classification of the spermatozoa between live (green fluorescence) and dead (red fluorescence) is done.
Analysis time is two seconds per field at 20x magnification

62. SCA® VITALITY for the automatic analysis of sperm viability

63. Generation of a spermiogram with medeaLAB Work with the database
Spermiogram: medea LAB offers the functionality to store up to 4 spermiograms for one patient. To change the actual spermiogram press button 1 – 4.Sample Volume: Enter the volume of the sample in this edit field. At the base of this value the concentration of the sample is calculated. Mean tracklength: The mean track length in this field. Result: The Results of the measurement for the actual field (Field) and the average value for all fields (Total). Sperm Conc. (Mio/ml): Concentration of the sample. This value is calculated on the base of the sample volume and the volume of the visible field of the optical system. Sperms Count: Number of processed objects. WHO a, b, c and d: Classification of the sperm movement due to the WHO-Criteria. See. Cap. 3.7.Motile, Local motile and Circle swimmer: Extensional Information of the sperm track classification. Open Track Window: Use the button open the actual track file. From this document you are allowed to analyse and export detailed track data.Fig The motility (spermiogram) database view
The motility (spermiogram) database view

64. Generation of a spermiogram with medeaLAB Work with the database
Field/Total Fields: Current field / Total number of fields .
Sperms Count: Number of processed sperms (Field and Total).
Normal Sperms, Pathological Sperms, Anomaly Head, Anomaly Midpiece und Anomaly Tail: Number of sperms belonging to the class.
Save Image: the current image is stored as an  JPEG-file with the name shown in the field on the right.
Learn Mode: Learn mode is activ.  All objekts which are classified manually are chosen to trainee the classificator.
Measure: Start a measurement.
Load Image: Loading a stored image
Database window to present the morphology data

65. Generation of a spermiogram with medeaLAB Work with the database
Carrying-out a morphology measurement
medeaLAB CASA is able to classify sperm cells automatically. But first the system has to be trained by manually assigning cells to classes in the Learn Mode. Thus a statistical classificator is trained, which then classifies objects automatically.
Steps to train the classificator of a sperm morphology sample:
Activate the “Learn Mode” on the database view.
Push the button “Measure
Morphology measurement overview

66. medeaLAB CASA Advantages
Generation of a complete spermiogram within a few seconds
Video recording and storage for re-analysis of sperm samples
Works on positive phase contrast, no special objective required
Accurate discrimination between sperm and non-sperm cells by multi-parametric filters (tail detector etc.)
High resolution mode for morphology analysis
Self-learning morphology analyzer, no adjustment of filters necessary
Analysis of external videos for inter-laboratory QC testing
MS Excel export of motility / concentration data for further analysis
Attractive price, reliable after-sales service
Hardcopy report layout in different languages

67. medeaLAB Concentration & Motility
Display / calculation of
Sperm concentration in mill/ml
Concentration of progressively motile sperm in mill/ml
WHO motility grades a, b, c, d in percent (old classification)
WHO motility progressive / local / immotile in percent (new classification)
VCL (curvilinear velocity) in µm/sec
VSL (straight-line velocity) in µm/sec
VAP (average path velocity) in µm/sec
WOB (wobble = VAP/VCL)
ALH (amplitude of lateral head displacement) in µm

68. medeaLAB Morphology Analyzer
Display / calculation of
Exact morphometry data for each individual sperm
Total number of analyzed sperm cells
% normal forms
% head abnormalities
% midpiece abnormalities
% tail abnormalities
% abnormalities of head and midpiece
% abnormalities of head and tail
% abnormalities of midpiece and tail
% abnormalities of head, midpiece
& tail
The Morphology Analyzer is self-learning, no manual adjustment of morphology classes or filters necessary

69. What is CASA What is CASA
CASA is used to study recorded video images of sperm through an extreamly powerful microscope. The images are then digitized and analyzed by a high-speed computer system (up to 60 frames per second) so that motility of individual sperm (and sperm populations) can be analyzed accurately. Why use Computer Assisted Semen Analysis?
The quality and accuracy of manual microscopic semen analysis varies enormously from center to center and despite efforts to introduce standardization and quality control, it remains a time consuming and poorly reproducible process. Hence reliability of such reports are questionable. Computer assisted semen analysis allows a very detailed determination of the parameters of sperm motility, the most important of which are:
VCL – Curvilinear Velocity – the total distance the sperm moves in a time unit (µm/s)
VSL – Straight Line Velocity (progressive velocity) – the distance the sperm moves along a straight line in a time unit (µm/s)
VAP – Path Velocity – mathematically smoothed path along which the sperm moves in a unit of time
ALH – Amplitude of Lateral Head Displacement (µm)
The CASA system is so precise that it enables our Andrologist’s to get a more accurate analysis by limiting the number of variables that exist with the manual analysis.

70. What is CASA

71. Why do we recommend CASA ?
Among numerous reasons why computer assisted semen analysis is worth choosing. Here are some of them:
Higher Reliability in predicting fertility – a CASA result correlates with the chance to become a father.
High degree of Precision, not provided by basic semen analysis
The test results are more Objective due to the standardization of testing conditions at each center
Motility parameters in CASA Reports are of greater usefulness in infertility evaluation in comparison with the results of standard semen analysis.
For those who believe in seeing for themselves, here’s how the CASA software screen looks like while analyzing the miniature miracle workers

72. Mira1000 Semen Analyzer (CASA)
MiraLab's Computer Assisted Semen Analysis System utilizes modern computer and advanced image processing techniques for clinical test of sperm motility, quality and morphology. Based on the sperm test standards of the WHO, the system can analyze the characteristic of sperm motions comprehensively through image processing of sperms in their dynamic or morphological status. The system then analyzes these values and generates accurate parameters to reflect sperms’ quality. The whole procedure is fast and provides several important information that are vital to present scientific basis for the male reproductive ability.
Color Test System
Deformity of sperm can be investigated and analyzed by the system by using papanicolaon's vaginal smear staining technique. The system can make the analysis process and report printing more clearly in addition to audio-visual documentation. Also the system has provided with the necessary conditions to standardize the elevation of the product for morphological analysis.
Wide Scope of Sperm Density Test without the Need of Sample Dilution
With the aid of powerful software function; the system can analyze more than a thousand sperms in one visual field at the same time. Sperm sample with a density of sperm/milliliter can be analyzed without dilution accurately, which facilitates (accelerates) the clinical examination greatly.

73. Mira1000 Semen Analyzer (CASA) (continue)
Creative Constant-Temperature Operation Desk
The operation desk makes provides a constant temperature of 37 during sperm testing during the whole test procedure. This in turn eliminates the influence of excessive low outside temperature on the tested sperm such as sperm velocity, sperm vitality etc.
Advanced Demarcating Function
The system can calibrate the magnifying rate automatically in order to ensure the consistency of the test result under different magnifying rates which improves the flexibility and adaptability of the tests of various magnifying rates.
Unique Virtual Grid Function
This function has realized the direct comparison of test results between human being and machine in the system. If you wish to use manual counting there is no need to buy an expensive calibrated counting chamber, it is all in your screen with the aid of virtual grid function
Strong Database Applying the Most AdvancedProgramming Language
The system can store a large quantity of patient records and image materials. This will provide detailed information for clinical research. Searching the database for specified cases has never been more easy and fast.
More Convenient Operation
Realizing the direct dialogue between human being and machine on the windows operation flat-top made it possible to avoid the over elaborate setting of parameters such the setting of various kinds of thresholds made the operation more convenient and easy even for rocky computer users.
Novel State of the Art Design
Patient now can see the motion of his own sperm directly through a monitor in the waiting room .The video can record and retain the dynamic image at any time with the aim of the analysis, diagnoses and the scientific research in the future.

74. The CEROS II sperm analyzer
Save Space, Save Money If you have limited bench space, a limited budget, or both – the CEROS II is the answer for your computer assisted semen analysis needs. Providing the same sperm analysis accuracy and detail of the IVOS II, the CEROS II is a cost-effective and space-saving alternative. Since the CEROS II processor is a separate unit from the optical component, the computer can be placed under the bench and out of the way. And, if you already own a compatible microscope (must be negative phase contrast trinocular), you can save even more money.
Components
The CEROS II is a component based system comprised of:
Multispecies Sperm Analysis Compatibility
Depending on your need, the CEROS II comes with Swine Breeders II, Equine Breeders II software, Animal Breeders II software or Animal Motility II software.
Ease of Use
The CEROS II hardware and software have been designed to provide users with a system that offers high-end performance while maintaining its ease of use. The intuitive Windows-based software promotes fast learning and quickly increases the confidence level of users. With a minimum investment of time, even users unfamiliar with computers will be performing fast, accurate and reliable sperm analyses. You don’t need to be a computer expert to analyze sperm with the CEROS II.
Sperm Analysis Results You Need
Whether you need basic counts and motilities or detailed analysis of sperm motion, the CEROS provides you with the results you need. Results for motile, progressively motile and static sperm include actual number counted, sample total, concentrations and percentages. Additional results include averages and distribution bar charts of velocities, motion characteristic and morphometry.Real-Time Quality Control
Just like the IVOS, the CEROS sperm analyzer provides on demand quality control with the Playback feature. Using image Playback, the accuracy of the analysis is confirmed. To optimize the system, analysis parameters may be adjusted using the two interactive QC Plots or directly from the analysis setup screen

75. IVOS Sperm Analyzer Integrated optics
IVOS Sperm Analyzer Sperm analyzer that integrates a built-in optical system, heated and automated stage, and high speed computer
Integrated optics
Automated, internal specimen stage
Multispecies sperm analysis compatibility 
Easy to use
Delivers the results you need
Real-time quality control
Rated moderately complex by CLIA
Summary of sperm analysis results provided, including counts, concentrations, mean values, and bar chart distributions

76. SQAIIC-P™ | A Standalone Semen Screening Device
Designed specifically for routine semen analysis screening in small and/or remote laboratories.  Includes a built in screen and on-board printer for reporting and documentation.  Designed for use with a simple disposable testing capillary. Assesses sperm concentration, motility, SMI and morphology in under 75 seconds.
Features:      Reports sperm count, morphology, motility and more.
     Requires no sample dilution      20 micro liter sample size options       High sensitivity mode or post vasectomy analysis      Self testing, self calibrating       Built in printer, disposable capillary  

77. QwikCheck™Gold | Fast Action in a Compact Package
Like its big brother, the SQA-V™ Gold, the QwikCheck™ Gold delivers 16 clinical parameters in 75 seconds with far greater precision and accuracy compared to the  manual method.  The system has an internal archive, optional label printer, and simple functionality.  Designed for smaller labs on a budget, this system is the perfect automated solution for routine fertility screening and sperm banking quality control. 
Features:      Reports sperm count, morphology, motility and 13 additional parameters
     Requires no sample dilution      .25ml and 20 micro liter sample size options       Runs fresh, washed, and frozen sample types      Runs latex bead or stabilized sperm quality control material      Self testing, self calibrating, External label printer, disposable testing capillary

78. Both CASA (computer assisted sperm analysis) and SQA-V (sperm duality analyzer) were easy to integrate into the laboratory routine and demonstrated an acceptable agreement versus manual semen analysis performed according to WHO 5th ed. manual. The best precision among all three methods was shown by the SQA-V system. Both automated systems, and particularly SQA-V, gave accurate morphology results. Automated sperm analysis systems can be considered accurate tools for routine sperm analysis, providing high quality results and allowing better standardization than manual analysis

80. Part V Semen parameters Counting Chamber
Special counting chambers with fixed depth should be used for motility analysis: Leja, Makler, MicroCell, Cell-Vu etc. Such chambers provide standard volume and monolayered distribution of sperms in one focal plane. counting chamber: reusable MMC-SK sperm counting chamber.Leja counting slides are useful for a Computer Aided Sperm Analysis System if the clinic or laboratory has got a large volume of analyses. Leja offers a wide range in disposable counting chambers for standard semen analysis. There are slides with 2 or 4 chambers and with chamber depths of 10 or 20 micron. The Leja slides have a standard low level of quantification which implies that even low sperm counts can be determined. The clinical implication of this is that you are well equipped to determine a threshold to choose between IUI, IVF, or ICSI for example.
  Microscope  
Trinocular upright transmitted light laboratory grade microscope to mount camera on it.
Camera adapter corresponding with image sensor size.

81. Computerized Semen Test
Sperm test for morphology The appearance of sperm under a microscope is used as a sperm test. All men, in a semen specimen, will have some sperm with a normal appearance and some with an abnormal appearance. In men with normal fertility, a certain percentage of the sperm will have a normal appearance..
Other types of male fertility tests
Endocrine (Hormone) Evaluation: Normal sperm production and sexual function are dependent on a normal hormonal environment. An endocrine evaluation should be performed if a sperm test reveals:
low sperm concentration
low sperm motility
The male hormone evaluation includes measurement of Follicle stimulating hormone (FSH)
Luteinizing hormone (LH)
Testosterone
Estradiol
Prolactin
Thyroid function
Liver function
Additional Semen Tests
They are no longer considered to be part of the standard male fertility test. Antisperm antibodies
Semen fructose – the absence of fructose, a sugar-like substance in the semen, means either the vas deferens are obstructed or that the seminal vesicles are absent.
Peroxidase staining – differentiates white blood cells from immature sperm to assess for possible infection.
Sperm penetration assay (Hamster egg penetration test)
Human zona pellucida binding test – measures the ability of sperm to bind to the zona pellucida (outer covering) of the egg. This test is also called the hemizona assay.
Sperm DNA fragmentation testing
Specialized Semen Tests
These tests may be useful in a small number of patients for under certain special circumstances
Hypo-osmotic swelling test – assesses the sperm membrane for structural integrity.
Semen culture – checks for bacteria that may cause genital infection.
Vital staining – determines numbers of living and dead sperm.
Genetic Evaluation
The importance of genetic evaluation in infertile males with severe oligospermia (sperm counts of less than 5 to 10 million per ejaculate) or nonobstructive azoospermia (absence of sperm in semen, not due to blockage) has recently been established. These patients may have abnormalities in the number of chromosomes (karyotype) or abnormalities in the structure of the male chromosome (microdeletion of the Y-chromosome). Patients with azoospermia as a result of being born without two vas deferens frequently have a mutation of a gene responsible for the disease cystic fibrosis but do not have the disease itself.

82. Computerized Semen Test
The American Society for Reproductive Medicine recommends that at least two sperm tests be performed in the evaluation of an infertile male. The semen is collected by masturbation on separate days are recommended. The ejaculate is collected in a sterile container provided by the clinic or laboratory and should be examined within 30 minutes of collection. Components of the semen test
Liquefaction (conversion into a liquid): complete within 60 minutes
Appearance: homogeneous, gray-opalescent ejaculate
Volume (amount): 2 milliliter or more
Concentration: 20 million per milliliter
Total count: 40 million sperm per ejaculate
Motility (movement): 50% of the total number of sperm
Progressive motility: 25% of the total number of sperm
Consistency: Not viscous (not thick)
Morphology (structure): More than 14% have a normal appearance
Semen tests became much more sophisticated with the advent of the computerized sperm tests. Computerized Semen Test For many years, evaluation of the semen has been performed by a laboratory technician looking through a microscope and manually counting sperm. It was completely arbitrary. Semen test with sperm motility analysis The determination of the number of sperm that are moving rapidly in a forward progressive manner is of paramount importance in the evaluation of male fertility. In an average semen test, there will be some sperm with no movement, some that are moving slowly and/or some that do not move in a forward direction. These sperm are less likely to be able to produce a pregnancy easily. With a computerized semen test, the computer identifies and tracks every sperm seen in under the microscopic field. Over a fraction of a second, the path the sperm has traveled is analyzed and many different parameters can then be computed with a high degree of accuracy. In the picture below, the squiggly green lines are the paths of individual sperm. The red dots mark sperm that are not moving.

83. Patient and semen parameters

84. Motility parameters The following parameters are calculated:
VCL  =  curvilinear velocity (micron/s). Time-average velocity of a sperm head along its actual curvilinear path, as perceived in two dimensions in the microscope.
VSL =  straight line velocity  (micron/s). Time-average velocity of a sperm head along the straight line between its first detected position and its last.
VAP  =  average path velocity (micron/s). Time-average velocity of a sperm head along its average path. This path is computed by smoothing the actual path.
LIN = linearity. The linearity of a curvilinear path.
STR straightness. The linearity of the average path.
BCF = beat cross frequency (beats/s). The average rate at which the sperm's curvilinear path crosses its average path.
ALH = amplitude of lateral head displacement. Magnitude of lateral displacement of a sperm head about its average path.
WOB = wobble. A measure of oscillation of the actual path about the average path, VAP/VCL.
Elongation of sperm head.
Mean values of all the parameters are represented in statistics.

85. The picture below, the squiggly green lines are the paths of individual sperm. The red dots mark sperm that are not moving
Semen test with sperm motility analysis

86. Normal sperm motility by CASA system

87. Circular sperm motility by CASA system

88. Semen test with sperm motility analysis
Some of the parameters that are calculated include:
Overall motility-Percent of sperm showing any movement
Rapid motility-Percent of sperm traveling at a speed of 25 um/sec or faster
Linearity-Percent of sperm moving in a straight line path
Progressive motility-Percent of sperm moving rapidly AND in a straight path
Mean velocities-An average speed for all sperm in the field of view
Amplitude of lateral head displacement-The average distance that the sperm head "wiggles" back and forth while moving
A computerized semen test will give many more parameters that are useful to the fertility specialist. Learn about the measurements of sperm motility
Here is a sample report looks like.

90. Morphology of the sperm
Morphology of the sperm head is an important criterion for the correct diagnosis. The software is set up to analyze still images of smears stained with the Diff-Quik stain according to strict Krueger’s criteria. We have selected Diff-Quik as worldwide recognized leader in rapid staining of sperm. With Diff-Quik, the head is stained pale blue in the acrosomal region and dark blue in the post-acrosomal region which is a good basis for precise image analysis. The following parameters are assessed for every spermatozoon:      
Area of the head.
FFC = form factor circle. The degree of similarity of the sperm head to a circle.
Perimeter of the head.
Brightness.
ELL_B = Big axis of ellipse outlining the sperm head, the length of the sperm head.
ELL_S = Small axis of ellipse outlining the sperm head, the width of the sperm head.
Elng = elongation of the sperm head.
FFE = form factor ellipse. The degree of similarity of the sperm head to an ellipse.
Acrosome = Percentage of the acrosomal region.
Mean values of all the parameters are represented in statistics.
The software classifies spermatozoa into Norm and Head Pathology classes automatically based on head parameters. You can easily correct the results manually and also specify other abnormalities (Tail Pathology, Neck Pathology).

92. Grading of sperm cells under high magnification

93. Sperm morphology.

94. Results of sperm morphology assessment

95. SpermMorph uses intelligent X,Y sampling and analyses more than 40 parameters on each sperm cell.

96. ORIGIN OF DNA DAMAGE IN SPERMATOZOA

97. OXIDATIVE STRESS AND EFFECT ON SPERM MOTILITY
Seminal ROS levels, when present in excess, possess potentially toxic effects on both sperm quality and function . Elevated seminal ROS production has been associated with decreased sperm motility, defective acrosome reaction, and loss of fertility . Sperm cell dysfunction, a result of ROS damage, is dependent on the nature, amount, and duration of exposure to ROS. The extent of ROS damage is also dependent upon surrounding environmental factors such as oxygen tension and temperature as well as the concentrations of molecular components such as ions, proteins, and ROS scavengers .

98. ROLE OF OXIDATIVE STRESS IN SPERM DNA DAMAGE AS RELATED TO MALE INFERTILITY

99. Statistics on all the parameters which have been measured in Motility and Morphology is displayed in the database (mean values of parameters). Statistics provides additional information on sperm quality. Corresponding database fields are filled in automatically during analysis like those for motility and morphology results. Statistics can be printed out to represent it to patient if required. Extended report template should be selected for this purpose (see below). Should the statistical data not be required for the patient, a brief report without statistics can be selected

100. Swimming Boys