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- •Foreword
- •Preface
- •Acknowledgments
- •Contents
- •Contributors
- •Introduction
- •Tissue Characteristics
- •Ovarian Scanning
- •Embryo/Fetus Susceptibility
- •References
- •Instrument Outputs
- •The Output Indices
- •Introduction
- •Limitations
- •History
- •Conclusions
- •References
- •Introduction
- •Endometrial Blood Flow
- •Ovarian Stromal Blood Flow by 2D Doppler
- •Ovarian Stromal Blood Flow by 3D Doppler
- •Conclusion
- •References
- •Transabdominal Ultrasound
- •Transvaginal Ultrasound
- •Postmenopausal Ovaries
- •Premenarchal Ovaries
- •Reproductive Age Ovaries
- •Antral Follicle Count (AFC)
- •References
- •Ovarian Cysts
- •Conclusion
- •References
- •6: PCOS
- •The Polycystic Ovarian Morphology (PCOM)
- •Ovarian Volume
- •Ovarian Stromal Blood Flow
- •Future Points
- •References
- •7: The Normal Uterus
- •Uterus
- •Myometrium
- •Endometrium
- •Cervix
- •References
- •8: Congenital Uterine Anomalies
- •Introduction
- •Müllerian Agenesis
- •Unicornuate Uterus
- •Uterus Didelphys
- •Bicornuate Uterus
- •Septate Uterus
- •Arcuate Uterus
- •Hysterosalpingography
- •Two-Dimensional Ultrasonography
- •Pelvic Magnetic Resonance Imaging
- •Three-Dimensional Ultrasonography
- •Urinary Tract Imaging
- •Conclusion
- •References
- •9: Uterine Fibroids
- •Background
- •Ultrasound
- •Saline Infusion Sonohysterography
- •Magnetic Resonance Imaging
- •Observation
- •Medical Therapies
- •Myomectomy
- •Hysteroscopic Myomectomy
- •Abdominal Myomectomy
- •Laparoscopic Myomectomy
- •Uterine Artery Embolization
- •MRgFUS
- •Conclusion
- •References
- •10: Uterine Polyps
- •Endometrial Polyps
- •Interrupted Mucosa Sign
- •Sonoelastography (SE)
- •Sonohysterography
- •Cervical Polyps
- •References
- •11: Intrauterine Adhesions
- •Introduction
- •Incidence
- •Manifestation
- •Causes
- •Risk Factors
- •Diagnosis
- •Hysteroscopic Surgery
- •Treatment Outcome
- •Radiographic Methods
- •Mechanical Barriers
- •Fluid Barriers
- •Tissue Barriers
- •Prevention Strategies
- •Recent Advances
- •Conclusion
- •Introduction
- •SHG Procedure [1, 2, 6, 13]
- •2D Versus 3D SHG
- •References
- •Gel Instillation SHG
- •SHG Versus Hysteroscopy
- •Conclusion
- •References
- •Introduction
- •Scrotal Ultrasonography
- •Paratesticular Structures
- •Epididymis
- •Varicocele
- •Vas Deferens
- •Testicular Ultrasound
- •Cryptorchidism
- •Cysts, Hydrocele, Infectious Processes
- •Testicular Masses
- •Microlithiasis
- •Testicular Torsion/Trauma
- •Transrectal Ultrasonography
- •Prostate
- •Cysts
- •Ejaculatory Duct Obstruction
- •Seminal Vesicles
- •Assisted Reproductive Techniques
- •Conclusion
- •References
- •Hysterosalpingography (HSG)
- •The Technique
- •Three-Dimensional Coded Contrast Imaging (3D CCI) During HyCoSy
- •Conclusion
- •References
- •Premature Luteinization
- •Multiple Pregnancies
- •Polycystic Ovarian Syndrome (PCOS)
- •Ultrasound Diagnosis [17]
- •Ovaries
- •Follicles
- •Clomiphene Citrate
- •Gonadotropins
- •Conclusion
- •References
- •Introduction
- •Normal Folliculogenesis
- •Monitoring Follicular Maturation
- •Standard Ultrasound Monitoring Program
- •Self-Monitoring
- •Conclusion
- •References
- •17: SonoAVC (Sonographic-Based Automated Volume Count)
- •Introduction
- •How Does One Apply SonoAVC?
- •Follicular Monitoring
- •Case 1
- •Case 2
- •Case 3
- •Antral Follicle Count
- •References
- •18: Ultrasound-Guided Surgical Procedures
- •Introduction
- •Uterine Septum
- •Submucosal Fibroids
- •Synechiae
- •Intrauterine Foreign Bodies
- •Hematometra
- •Summary
- •Ovarian Cyst Aspiration
- •Hydrosalpinx Aspiration
- •Oocyte Retrieval
- •Endometrial Thickness
- •Embryo Transfer
- •Conclusion
- •References
- •References
- •Introduction
- •Clinical Touch ET Versus Transabdominal US-Guided ET
- •Conclusion
- •References
- •General Concepts
- •Patient’s Acceptance
- •Contraindications
- •Radiation
- •Image Post-Processing
- •Conclusion
- •References
- •Introduction
- •A Quick Look Back at Endometrial Assessment Approaches
- •Receptive
- •Non-receptive
- •Improving Endometrial Receptivity Assessment
- •References
- •List of Relevant Websites
- •23: Early Pregnancy Ultrasound
- •Introduction
- •Pregnancy Location
- •Gestational Sac (GS)
- •Yolk Sac (YS)
- •Embryonal Heart Rate (EHR)
- •Pregnancy Dating
- •Pregnancy Viability
- •Conclusion
- •References
- •24: Ectopic Pregnancy
- •Cervical Pregnancy
- •Ovarian Pregnancy
- •Abdominal Pregnancy
- •Cesarean Scar Ectopic Pregnancy
- •Interstitial Ectopic Pregnancy
- •Ectopic After Hysterectomy
- •Summary
- •References
- •Index

15 Ultrasound inFollicle Monitoring forOvulation Induction/IUI
269
is suggested that hMG/hCG administration
should be interrupted in the presence of 11 or
more preovulatory follicles, especially if most of
them are immature (<9mm).
Conclusion
Ultrasound is the most powerful tool to monitor
normal and stimulated cycles; predictions of the
assumed time of ovulation allow optimal timing
of various procedures such as insemination,
ovum aspiration, etc.
In stimulated cycles sonographic detection of
too many follicles allows withholding hCG
induction thus preventing hyperstimulation.
In the past ovulation function was monitored
by estradiol estimation; since the development of
sophisticated ultrasonographic techniques, monitoring of ovarian follicular growth by ultrasound
became a routine addition to estradiol measurement in most clinics.
Accumulating data based on the Cochrane
Database [38] indicate that there is no evidence
from randomized trials to support cycle monitoring
by ultrasound plus serum estradiol as more efcacious than cycle monitoring by ultrasound only on
outcomes of live birth and pregnancy rates.
As far as OHSS, randomized trial with a sufciently large sample is needed. Until such a trial
is considered, ultrasound plus serum estradiol
may need to be retained as a precautionary good
practice point, in patients prone to develop
hyperstimulation.
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2D Ultrasound inFollicle
Monitoring forART
GianlucaGennarelli, TomerTur-Kaspa,
AlbertoRevelli, MetteToftager,
andDavidP.Cohen
16
Introduction
Ultrasound (US) imaging represents an invaluable
tool for the reproductive endocrinologist working
with assisted reproductive technologies (ART). It
allows for noninvasive monitoring of the ovarian
response during controlled ovarian stimulation
(COS). Ultrasound imaging of follicle maturation
was rst performed in 1978, when a linear correlation between follicle size and serum estradiol
levels was demonstrated [1]. Later, studies performed in the early 1980s conrmed the relationship between serum estrogen concentration and
both the number and size of growing follicles. An
increase in uterine size and endometrial thickening during stimulation was also described [2].
Currently two-dimensional (2D) ultrasound imaging is used diagnostically and in treatment, for
evaluation of all pelvic organs and for transvaginal procedures, respectively.
G. Gennarelli · A. Revelli
Sant’Anna Hospital, University of Turin, Department
of Obstetrics and Gynecology, Turin, Italy
T. Tur-Kaspa
Wesleyan University, Middletown, CT, USA
M. Toftager
Fertility Clinic, Rigshospitalet, Copenhagen
University Hospital, Copenhagen, Denmark
D. P. Cohen (*)
Institute for Human Reproduction, Chicago, IL, USA
e-mail: dcohen@infertilityihr.com
Most importantly, the introduction of transvaginal ultrasound in 1983 dramatically improved the
safety and success of ART. Today, transvaginal
ultrasound imaging is imperative when administering gonadotropins for COS, in order to optimize
treatment, reduce the risk of multiple pregnancies,
and avoid potentially life- threatening side effects,
such as the ovarian hyperstimulation syndrome
(OHSS). Indeed, the use of 2D ultrasound for
assessing follicular development during gonadotropin stimulation for ART is virtually universal.
More recently, the addition of power Doppler
added to 2D ultrasound has been proposed with
the aim of studying ovarian and endometrial
blood ow. By monitoring perifollicular blood
ow, the physician can identify follicles whose
oocytes may result in embryos of better developmental competence [3] (see Chaps. 3 and 4).
Why Monitor theFollicular Phase?
During in vitro fertilization (IVF) treatment,
exogenous gonadotropins induce the growth of a
cohort of ovarian follicles. Monitoring this phenomenon is essential in order to optimize and
individualize IVF treatments, and ultrasound
examination is effective at all stages of the COS.
At a baseline examination, before initiating
gonadotropin administration, US monitoring can
identify ovarian abnormalities, such as follicular
cysts or adnexal masses that need to be addressed
© Springer Nature Switzerland AG 2019
L. A. Stadtmauer, I. Tur-Kaspa (eds.), Ultrasound Imaging in Reproductive Medicine,
https://doi.org/10.1007/978-3-030-16699-1_16
273

274
G. Gennarelli et al.
in advance. Furthermore, a baseline count of small
antral follicles (AFC) provides a hint into the
probability of success, and it helps identify
the optimal starting dose of gonadotropin and
the most appropriate type of stimulation protocol.
It has been repeatedly shown that the AFC correlates positively with the number of oocytes
retrieved, the proportion of mature oocytes from the
oocytes retrieved, and ultimately the probability of
success of the IVF treatment [4, 5], provided a
transvaginal US probe with frequency ≥7MHz is
used and the operator is adequately experienced [6].
Furthermore, in cases when GnRH agonists are
used to block the endogenous gonadotropin secretion from the pituitary, US examination can verify
that effective pituitary-ovarian axis suppression has
been obtained before starting COS. Additionally,
other useful information can be obtained before
stimulation, for example, the ease with which the
ovaries are accessible to transvaginal needle aspiration. Finally, during ovarian stimulation US monitoring is crucial in order to determine whether the
initially suggested gonadotropin stimulation dosing protocol is optimal or needs to be adjusted.
As previously mentioned US helps predict the
number of oocytes expected to be obtained, but it
is also of paramount importance for minimizing
the risk of OHSS.When OHSS is incipient, free
uid may be identied in the peritoneal cavity.
This sign, together with the number and size of the
growing follicles and the ovarian volume, is information currently used to help predict and prevent
OHSS.Interestingly, to prevent OHSS in the polycystic ovarian syndrome patient resistant to standard ovulation induction, ultrasound has been
suggested as a tool to guide ovarian drilling [7].
Finally, ultrasound monitoring is critical in
determining the optimal time for induction of the
nal maturation of the growing follicles. As a
matter of fact, most clinicians agree that ultrasound examination is the best modality, among
those available, to monitor ovulation induction.
which occur during normal folliculogenesis.
Follicles grow through two stages, the rst being
gonadotropin-independent and the second
gonadotropin- dependent. Primordial follicles
consist of an oocyte with a thin layer of granulosa
and stromal cells, and at this stage, follicles cannot be seen by ultrasound. By the time follicles
develop an antral cavity, they become ultrasonographically visible, and, importantly, they have
reached the gonadotropin-dependent stage. These
antral follicles measure between 2 and 10mm in
mean diameter and represent the pool of follicles
that may be stimulated in the ensuing follicular
phase. In a natural cycle, one follicle is ultimately
selected for ovulation, and that selection process
occurs during the mid-follicular phase of the
ovarian cycle, when the endogenous pituitary follicle stimulating hormone (FSH) level falls in
response to the increasing ovarian estradiol production. Decreasing FSH levels promote a selection process in which each of the follicular
microenvironments competes for the diminishing
FSH needed to stimulate granulosa cells to produce aromatase and continue growth. Aromatase,
in turn, is necessary to convert testosterone and
androstenedione produced in the peripheral theca
cells into estradiol and estrone, respectively, in
the granulosa cells. Failure of this conversion
from androgen to estrogen leads to an elevated
androgen-to-estrogen ratio in the microenvironment of the follicle and then to follicular atresia.
From this cursory review of the anatomy and
physiology of oocyte maturation, it is easy to see
how ovulation-inducing agents that either indirectly increase endogenous FSH (e.g., clomiphene citrate) or directly add exogenous FSH to
the system diminish the competition among follicles for FSH and permit the development of
multiple dominant follicles [5, 6].
Monitoring Follicular Maturation
Normal Folliculogenesis
Before reviewing the parameters of follicular
growth studied with 2D ultrasound during ART
treatment, it is helpful to summarize the events
Methods forMonitoring
It is difcult to predict the optimal number of
growing ovarian follicles in an IVF cycle, since
there is considerable variation in ovarian response
among patients. The ovarian response depends on

16 2D Ultrasound inFollicle Monitoring forART
275
age, ovarian reserve, the type of stimulation protocol, the FSH dose, FSH receptor polymorphisms, etc.
Follicular maturation in IVF cycles can be
monitored clinically by:
• Serum estradiol value alone
• 2D ultrasound alone
• 3D ultrasound alone
• Serum estradiol and ultrasound combined
• Supplemental power Doppler imaging
There are numerous studies on the use of
these different methods for monitoring follicular
maturation. Traditional monitoring of an IVF
treatment cycle includes a combination of 2D
ultrasonography and serum estradiol concentrations and has long been accepted as the gold
standard. However, whether estradiol monitoring adds any advantage to US monitoring
remains controversial. Obviously, US examination provides more accurate measurement of follicle number and size than can be obtained by
serum estradiol alone. Whereas available data
are not conclusive, the most recent literature
does not support the notion that measuring estradiol serum concentrations during COS is of signicant advantage in terms of both results and
safety [8].
The frequency of US check and the time
points at which the dose of exogenous gonadotropins could be changed show wide variations,
depending on the experience of the clinician and
the routine applied at each IVF clinic. Some
monitoring methods are very complex, whereas
other methods are simpler. However, whatever
the complexity of COS monitoring, the outcomes
of IVF cycles does not differ signicantly [9].
When viewed on ultrasound, follicles appear
as echo-free structures within the more echogenic ovarian tissue. By convention, follicle size
in 2D is estimated by calculating the mean of
the maximum follicular internal diameter in two
perpendicular planes [10]. Alternatively, the follicle size can be estimated in three dimensions,
the x, y, and z planes, and using this technique,
it is possible to calculate the volume of each follicle. Most recently, three-dimensional (3D)
software programs that distinguish the echo-
genicity of the well-circumscribed, sharp-edged,
echolucent follicular uid from the surrounding
greater echogenicity of the ovarian cortical
parenchyma have automated this process and
allow follicular volume calculations from data
derived from 2D images. The clinician can simply sweep through the ovarian tissue, and the
stored image data is analyzed, reducing the time
usually needed when multiple diameters are
measured for each follicle, separately. This
option is particularly useful in busy practices,
when a large number of follicular monitoring
scans are performed every day.
Standard Ultrasound Monitoring Program
Follicular growth can be directly monitored with
2D ultrasound, since the follicular diameter
increases during development in the follicular
phase. Most clinicians measure the follicles at
baseline (Fig.16.1), prior to initiating gonadotropin stimulation, and then again after approximately 5days of gonadotropin stimulation, and
then every 24–48hours depending on the rate of
development (Figs.16.2, 16.3, 16.4, and 16.5).
Once the mature follicle measures 17–21 mm,
the physician can trigger ovulation with human
chorionic gonadotropin (hCG, a luteinizing hormone (LH) surrogate) (Fig. 16.6) or a
gonadotropin- releasing hormone analog. In simple ovulation induction, conrmation of ovulation can be demonstrated by ultrasound as well,
observing the sudden change of the intact follicle, made up of concentric layers of theca cells
surrounding granulosa cells enclosing the follicular uid and the oocyte that is suddenly changed
after follicular rupture and ovulation.
Physiologically, it is at this moment that both
testosterone-secreting theca cells and
estradiol- secreting granulosa cells convert intracellular steroid production to preferentially favor
progesterone production. Thus, ovulation is
accompanied by a dramatic loss of the concentric
architecture of the preovulatory follicle and an
increase in blood supply, presumably aimed at
increasing dramatically the output of progesterone into the bloodstream. A very specic, nearly

276
Fig. 16.1 Baseline,
prior to initiating
gonadotropin
stimulation. Ovary with
antral follicles
Fig. 16.2 Stimulation
day 5, showing recruited
follicles measuring
10–12mm
G. Gennarelli et al.
Fig. 16.3 Stimulation
day 7, showing ovary
with leading follicle
>12mm

16 2D Ultrasound inFollicle Monitoring forART
Fig. 16.4 Stimulation
day 9, showing ovary
with growing follicles
Fig. 16.5 Stimulation
day 11, 2–3 follicles
measuring 17–18mm
277
Fig. 16.6 Day of
ovulation induction.
Leading follicles
measuring more than
18mm

278
Fig. 16.7 Corpus
luteum cyst– “ring
of re”
G. Gennarelli et al.
pathognomonic “ring of re” ultrasound nding
is easily discerned around each corpus luteum at
this time (Fig.16.7). Doppler technology added
to the 2D image enables observation of this typical ow pattern which, in combination with the
classic echogenicity of the corpus luteum, makes
the diagnosis of ovulation quite simple [11].
Follicular Size andVolume
During COS for IVF, it is realistic to recruit 5–10
ovarian follicles in each ovary; however the number, the rate of growth of each follicle, and the
number of stimulation days can vary greatly
between patients.
After 6–7 days of gonadotropin stimulation,
follicles measuring more than 10mm are expected.
Once a dominant follicle measures more than
12mm in mean diameter, a growth rate of 2mm
(1–3mm) per day is expected [12]. Growth continues until follicular maturation at 17–21mm,
and at that point, the oocyte is ready to ovulate,
complete meiosis in response to the LH trigger,
and be released into the peritoneal cavity.
oocytes to be fertilized; mature oocytes are dened
as those that have completed meiosis I, extruded
the rst polar body, and rearrested in metaphase of
meiosis II.Most commonly hCG is administered
to mimic the endogenous LH surge, which in turn
triggers meiotic reinitiation from the oocyte’s prophase I resting state. As noted previously, stimulation protocols vary and are often modied during
the stimulation. However, in most cases ovulation
is triggered when ≥3 follicles ≥17mm in diameter
are identied on ultrasound. Alternatively, the trigger is administered either when ≥3 follicles, each
with a maximum diameter of 18mm, are identied
or when ≥1 follicle of ≥18mm and 3 follicles of
≥15mm are observed. Other criteria have taken
into consideration serum estradiol levels as well;
in these criteria, hCG is usually administered when
the leading follicle reaches 18–20 mm and the
coincident serum estradiol level suggests “satisfactory” follicular development. More in detail, it
is suggested that induction of the nal oocyte maturation be performed in the presence of at least
one follicle ≥20mm and a serum estradiol level
≥1200pg/mL [13]. So far, there are no conclusive
data in favor of any specic criteria.
Criteria Used forTriggering Ovulation
Whereas the criteria used for triggering ovulation
and inducing the nal oocyte maturation vary
between protocols, all aim to produce mature
How toPredict Retrieval ofMature
Oocytes?
Follicular size and the volume of follicular uid
have traditionally been recognized as possible

16 2D Ultrasound inFollicle Monitoring forART
279
predictors of oocyte quality, i.e., oocytes that will
be fertilized and result in embryos that implant
and progress into a viable pregnancy [14].
However, attempts to nd a universally accepted
threshold of a worth-to-be-punctured follicle size
have been disappointing, due to conicting
results. What is commonly accepted is that a
large follicle is more likely to lead to the retrieval
of a mature oocyte than a smaller follicle. The
correlation between the follicle size and the likelihood of retrieving a mature oocyte may reect
the notion that larger follicles have completed the
maturation process and released the oocytecumulus cell mass as a free-oating structure in
the antral uid just before follicle rupture.
According to early data from Teissier etal. [15],
14 mm in diameter should be considered the
threshold follicle size to get an acceptable chance
of nding meiotically competent oocytes at
retrieval, both in normal and polycystic ovaries.
However, follicles measuring as small as
11–15mm have been reported to have as much as
a 50% chance of yielding a mature oocyte [16].
Conversely, follicles with a mean diameter
above 22mm result in lower recovery of mature,
fertilizable oocytes as they often contain postmature eggs, probably the result of intrafollicular
atresia and degenerative phenomena [17].
More recent studies have observed a correlation between oocyte competence and ultrasoundmeasured follicular size prior to trigger and
oocyte retrieval. A higher proportion of immature
oocytes (germinal vesicle-GV-stage) are indeed
found in smaller follicles, particularly those
below 12mm in mean diameter. It is important to
remember, however, that this is not a universal
nding since even small follicles can generate
mature MII oocytes [18]. Wittmaack et al. [19]
found the optimal follicle volume to be between
1ml, which corresponds to 12mm in mean diameter, and 6–7 ml, corresponding to 24 mm in
mean diameter. Higher oocyte recovery rates,
higher fertilization rates, and higher cleavage
rates for follicles in this interval were reported.
However, within this range, the higher the follicular volume, the higher the fertilization rate and
the better the embryo quality. Furthermore, in
sync with those observations, a recent study
showed that delaying the oocyte maturation trigger by 1 day, once the leading follicles have
reached 18 mm in diameter, would allow the
retrieval of signicantly more mature oocytes,
provided no progesterone rise is detected [20].
Most compelling, the results of a prospective
study, including 9933 follicles from 535 IVF
cycles, conrmed that oocytes from follicles with
a volume <1ml (<12mm in mean diameter) had
a signicantly lower fertilization rate than
oocytes from larger follicles. However, the same
oocytes, once fertilized, yielded embryos of comparable quality; no signicant differences in the
implantation rate, clinical pregnancy rate, or live
birth rate were detectable from embryos derived
from oocytes recovered from either small or large
follicles [21].
It is also true that in general, it is not possible
to identify a clear relationship between follicle
size and the morphological quality of the embryos.
This may be due to the fact that follicles with a
volume within a certain interval contain oocytes
that lead to embryos of comparable morphologic
scores and/or to the fact that the male gamete contributes to embryo quality as well and should be
considered along with the oocyte.
Likewise, follicular size, volume, morphology, and vascularity do not provide any predictive information on the chances of conception of
a euploid fetus.
The Uterine Cavity andMonitoring
ofEndometrial Proliferation
During the diagnostic evaluation of the infertile
couple, assessment of the uterine cavity and the
patency of the fallopian tubes is essential. The
standard means of assessment of these two structures is hysterosalpingography or more invasive
laparoscopy and hysteroscopy. In the last decade,
the use of saline infusion into the cavity to assess
both the cavities in several planes using 2D and
3D (more recently) transvaginal imaging has
made assessment of the uterine cavity and the
tubes an ofce procedure that is less painful,
involves no radiation, and is less expensive and
more accessible.
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