andrology and embryology

smen parameters for seminogram - lower references

§Semen Volume   - 1.5 ml                                

Sperm Concentration - 15 × 10^6 / ml

Total Sperm count - 39 × 10^6

Motility (Progressive Motility) - 32%

Sperm Morphology - 4%

Sperm vitality - 58%

all parameters greater lower references arw

normozoospermic - normal sperm

semen volume

beloq 1.5 ml - hypospermia

above 5.5 ml - hyperspermia

Measure of prostate and other glands secretions

Semen is essential for sperm protection, nutrition and survival

wet prep

looks at agglutination, debris and round cells

severe agglutinination suggests prescence of anti sperm antibidoes

10 micro litre sample

when doing a wep prep why should you wait 1 minute after putting on the coverslip

to stop drifiting

• progressive motility

• assess around 200 sperms

sperm concentration

Using improved Neubauer haemocytometer

Counting at least 400 sperms

< 15 x 10⁶ / mL Oligospermia - fewer cells than normal

total sperm count + formula

• conc x volume

• LOWER REFERENCES

  ≥ 39 x 10⁶

  < 39 x 10⁶  Low sperm count

  < 1-2 x 10⁶  Azoospermia

Centrifugation of semen sample, < 1-2 x 10⁶  Azoospermia

morphology

abormal forms - globozoospermia

vitality - active

§Eosin-Nigrosin Stain

10ul Stain solution + 10ul Semen

ÞFeathering technique

Þ1000X

§

Dead sperm = permeabilised membrane

                        = Pink

Percentage of live sperm

semen smearing method

§Feathering method (a) & Pipette method (b)

Ethanol fixed and air-dried

sperm morphology staining

§Papanicolaou Stain

§

Head : Pale blue

Mid-Piece: Red staining

Tail: Blue

Excess cytoplasm: Pink

round cells

§Papanicolaou Stain

§

8: Epithelial cells

12: Macrophage

13 / 27 : Leukocyte (WBC)

22: Bacilli

26: Spermatid (95% of round cells)

Giemsa Stain for Leukocyte determination

sperm prep techniques §

§Removing abnormal sperm, debris and rounds cells

§Techniques:

ÞSwim-up

ÞDensity gradient (40% Upper / 80% Lower)

conc and mobility assessed

HIV Infected semen sample

§If HIV, viral RNA and proviral DNA are detected in sample: semen and non-sperm cells

§

§CD4, CCR5, CXCR4 are expressed only by non-sperm cells.

§

§Density gradient followed by swim-up as a way of preventing infection of uninfected female partners.

§

§Prepared samples should be tested via RT-PCR before use.

§

§Procedure should be carried out in secure facilities to minimise cross-contamination of HIV-free samples

early cleavage grading

x,x,x,

Number of cells, Blastomere size, fragmentation

blastomere size

fragmentation

Fragmentation

Due to uneven division of the cells

Cytoplasmic debris

Higher fragmentation = lower likely hood of pregnancy

blastocyst grading

blastocyst grading - 3BB minimal

Discards grade ‘C’ blastocysts = potential?

Single embryo transfer of expanded blastocysts with grade ‘C’ ICM or grade ‘C’ TE resulted in live births at rates that, while lower than top quality blastocysts (34.1 versus 46.8%), resulted in 109 live births that had similar obstetric and perinatal outcomes compared to grade ‘A’ or ‘B’ blastocysts

embryo transfer , what jhappens to other good quality embryos

1 or 2 good quality embryo transferred

Freezing

Other lower quality embryos: either discarded or if consented by parents, given to research project (NHS and HFEA ethical approval)

frozen embryo transfer

Thawed (+ in vitro developed)

Re-freezing = Twice-frozen-thawed embryos have a lower post-thaw survival rate

But similar pregnancy and live birth rates to once-thawed embryos