This document discusses several potential technologies for identifying viable oocytes, including polar body biopsy, spindle imaging, and zona pellucida birefringence. Polar body biopsy and array comparative genomic hybridization can be used to assess chromosomal disorders in oocytes. Spindle imaging using polarization microscopy can identify the meiotic stage of an oocyte. Zona pellucida birefringence imaging using polarization microscopy provides information on cytoplasmic maturity, with more structured zonae correlating with better developmental potential. These technologies, especially polar body biopsy, spindle imaging, and zona imaging, may help select the most implantation-competent embryos.

1. Other potential technologies to
identify the most viable oocytes:
polar body biopsy, spindle
imaging, Zona pellucida
birefringence
Markus Montag, Ph.D., Prof.
University of Heidelberg
Markus.Montag@med.uni-heidelberg.de

2. Polar body biopsy and array-CGH:
the impact of chromosomal disorders
Test - DNA
CONTROL- DNA
COT 1-DNA
Software-based
evaluation

3. Maternal age and aneuploidy rates
• Geraedts J, Montag M, Magli C et al. Polar body array CGH for prediction of the status of the
corresponding oocyte. I. Clinical results. Hum. Reprod. 2011, 26(11):3173-80.
• Magli C, Montag M, Köster M et al. Polar body array CGH for prediction of the status of the
corresponding oocyte. I. Technical aspects. Hum. Reprod. 2011, 26(11):3181-5
0
10
20
30
40
50
60
70
80
90
100
Age < 30 Age 31 - 340 Age > 40
Aneuploidyrate in stimulated cycles

4. Spindle imaging by polarization
microscopy: visualizing the invisible
Is this a metaphase-II-oocyte?

5. Spindle dynamics is physiological
Spindle bridge
MI -> MII
75-90 min
MII
40-60 min

6. Zona imaging by polarization microscopy:
a parameter of cytoplasmic maturity
Zonaimage – Cytoplasmic Maturity
Zona = product of the oocyte
Optimal maturation = good structured zona

7. Zona imaging: clinical use
0,0
10,0
20,0
30,0
40,0
50,0
60,0
70,0
HZB/HZB HZB/LZB LZB/LZB Kontrolle
*
#
% 20 50 65 68 cycles
LZB/LZB versus HZB/LZB and HZB/HZB
* P < 0.005
# P < 0.025
Pregnancy rate
Implantation rate

8. Zona imaging:
the molecular background
Recommended reading: Assou et al., Hum Reprod Update 2011
Pregnancy No pregnancy
Seven significantly differential expressed genes
in cumulus cells
Assidi, Montag, van der Ven, Sirard; JARG, 2011

9. The viable oocyte
• Primary target: oocyte maturity / follicular background
• Methodology: Polarization microscopy
o Spindle imaging for meiotic maturity
o Zona imaging for developmental competence
• Secondary target: oocyte aneuploidy
• Methodology:
o Polar body biopsy (PB1+2) and array-CGH
• Oocyte viability assessment is a valuable add-on for
subsequent selection of the most implantation-
competent embryo

10. Further reading
• Ebner T, Balaban B, Moser M et al. Automatic user-independent
zona pellucida imaging at the oocyte stage allows for the prediction
of preimplantation development. Fertil Steril, 2010;94:913–20.
• Madaschi C, Aoki T, de Almeida Ferreira Braga DP et al. Zona
pellucida birefringence score and meiotic spindle visualization in
relation to embryo development and ICSI outcomes. Reprod
Biomed Online 2009;18:681–6.
• Madaschi C, Aoki T, de Almeida Ferreira Braga DP et al. Zona
pellucida birefringence score and meiotic spindle visualization in
relation to embryo development and ICSI outcomes. Reprod
Biomed Online 2009;18:681–6.
• Montag M, Köster M, van der Ven K, van der Ven H. Gamete
competence assessment by polarization microscopy. Hum Reprod
Update, 2011;17:654-66.