1. Time-Lapse
-Cleavage-stage development is a dynamic process in which embryo
morphology may change significantly over a time span (h).
-Conventional grading practices may not detect subtle differences between
individual embryos, such as the time to progress from one cleavage division
to the next.
-The use of an automated instrument with programmable time-lapse image
acquisition, allows data to be collected for individual embryos during
development to quantify the exact timing of each cell division.

2. 5
15
25
35
45
Day 3Embryo CategoriesASEBIR
%Inside
%Outside
%Inside 33,2 17,1 20,4 29,4
%Outside 35,6 15,7 20,6 28,1
A B C D
p=0,941

3. Exact timing by time-lapse
Precise definition of exact timings (n=9530)
Variable Media
CI 95%
Minimum Maximum
Lower limit Upper limit
t2 27,93 27,88 28,11 18,40 74,45
t3 38,21 38,07 38,35 19,27 109,45
t4 40,71 40,56 40,85 20,24 94,4
t5 51,03 50,83 51,22 20,29 112,46
t6 54,06 53,88 54,25 25,17 115,6
t7 56,72 56,52 56,92 27,21 116,21
t8 59,11 58,86 59,35 33,37 116,21
Morula
86,63 85,85 87,41 59,95 99,65
Blastocyst
104,12 102,72 104,26 79,94 136,62
ICSI ~ 14:00h
D1 17:56h
D2
04:13h
06:43h
17:00h
20:00h
22:43h
D3 01:07h
D4
04:38h
22:07h

5. Implantation Success; ESDMorphology
included
ok
Grade A Grade B Grade C Grade D Grade E Discarded
non viable
excluded
yes no
yes nono yes
A+ A E+ EB+ B C+ C D+ D
CC2 5- 12h CC2 5-12h CC2 5-12h CC2 5-12h CC2 5-12h
yes no yes no yes no yes no yes no
Exclusion
Criteria
Direct Cleavage
Uneven Blastomere
T5
48-56h
T3
35-40h
T3
35-40h
embryo kinetics and implantation

7. Time-lapse + respiration
Respiration Peak = Citokinesis
Time-hours post ICSI
Embryo respiration and Implantation
5
5,2
5,4
5,6
5,8
6
6,2
6,4
6,6
6,8
7
0 1 2 3 4 5 6 7 8 9 10
fmol/sec
Cleavage O2 consumption pattern
Not Implanted
Implanted

8. Cumulus cells — Transcriptomics at an early stage
- To identify reliable genomic biomarkers
expressed in cumulus cells that accurately
and non-invasively can predict the oocyte
developmental competence and reinforce
the already used morphological criteria.
- Gene expression assessed using microarray
platforms
- The oocyte triggering stimulates the
expression of genes involved in the
granulosa luteinization process and
ovulation finality − a crucial step during
follicular development to produce a fully
healthy oocyte
- Several genes are also differentially
associated to successful pregnancy and
implantation

9. Investigating spent culture medium
Pyruvate
Glucose
Amino acids
Sugars
Uptake
Hormones
Lactate
Ammonia
Enzymes
Production
Other factors
- A noninvasive metabolomic profile of the culture medium surrounding the
embryo with an analysis of the components produced or depleted by
them can be achieved by different technologies.
- Multivariable analysis could be applied and models based on viability
index may be used in the embryo selection process.

10. Proteomics
• Multiple protein analysis of
human embryo secretions
offers the chance to expand our
perspective of a noninvasive
quantification of human
embryonic viability and even
chromosomal normality
avoiding any type of
manipulation.