Table Of ContentAshok Agarwal · Sajal Gupta
Rakesh Sharma Editors
Andrological
Evaluation
of Male Infertility
A Laboratory Guide
123
A ndrological Evaluation of Male Infertility
A shok A garwal • S ajal G upta • R akesh S harma
E ditors
A ndrological Evaluation
of Male Infertility
A Laboratory Guide
E ditors
A shok A garwal, PhD, HCLD (ABB), ELD (ACE) S ajal G upta, MD (Ob/Gyn), MS (Embryology),
A ndrology Center and Reproductive Tissue Bank TS (ABB)
American Center for Reproductive Medicine A ndrology Center and Reproductive Tissue Bank
Cleveland Clinic American Center for Reproductive Medicine
C leveland, O H, U SA Cleveland Clinic
C leveland, O H, U SA
R akesh S harma, PhD
A ndrology Center and Reproductive Tissue Bank
American Center for Reproductive Medicine
Cleveland Clinic
C leveland, O H, U SA
I SBN 978-3-319-26795-1 ISBN 978-3-319-26797-5 (eBook)
D OI 10.1007/978-3-319-26797-5
L ibrary of Congress Control Number: 2016941225
© Springer International Publishing Switzerland 2 016
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F oreword
R evolutionary advances in the diagnostic and therapeutic procedures for the management of
male infertility that foreshadow a growing role of andrology testing are the future of this fi eld.
Millions of men are evaluated yearly across the United States with a growing variety of fertility
tests. The days of the simple Makler chamber-generated semen parameters have been supple-
mented with computer-assisted semen analysis, as well as advanced semen tests such as oxida-
tive stress and DNA fragmentation assessments. Increasingly, these tests are allowing improved
diagnostic accuracy and stratifi cation to appropriate treatment options.
T his state-of-the-art laboratory manual contains protocols that can be used daily in the
investigation of male infertility. In its 24 chapters, the book covers most of the common routine
and advanced testing that is available at the present time. This text is unique in its emphasis in
presenting test protocols in detail that allow immediate use in a clinical laboratory setting. It
has a number of hand-drawn, artistic presentations of key equipment and procedures which
add to the clarity. At the beginning of the text, information is presented to place clinical aspects
of male infertility in context of the role of semen testing. It contains important chapters on
quality management, competency assessment, licensing, and accreditation which allow a lab
to provide the highest quality of testing while remaining compliant with applicable laboratory
standards. The protocols are arranged from the routine to the advanced laboratory procedures
in daily use. It emphasizes simple, concise protocols which can be readily transferred into
practice. The information on assisted reproduction-associated procedures including prepara-
tion for IUI specimens and cryopreservation techniques represents time-tested best practices
from one of the busiest andrology laboratories in the country.
D rs Agarwal, Gupta, and Sharma combine their over 50 years of experience in running a
state-of-the-art clinical andrology laboratory to provide an indispensable resource for thou-
sands of clinicians, reproductive professionals (andrologists, embryologists), laboratory tech-
nicians, as well as other students of andrology lab testing. This experienced team has authored
dozens of texts related to all aspects of male fertility. Their current book is a one of a kind
work—a must-read for all with an interest in providing the highest quality and most accurate
andrology testing for their patients.
In the growingly complex world of sperm testing, they have provided a simple, elegant
resource. It will be a welcome addition to your bookshelves, and I expect you will fi nd it valu-
able on a regular basis. I hope you will enjoy it!
S eptember 25, 2015 E dmund S abanegh J r., MD
Department of Urology
The Cleveland Clinic
Cleveland, OH, USA
v
C ontents
1 D iagnostic Tests in the Evaluation of Male Infertility .......................................... 1
A hmad M ajzoub and E dmund Sabanegh Jr.
2 Q uality Management in Andrology Laboratory ................................................... 1 1
E rma Z . D robnis
3 C ompetency Assessment in Andrology Laboratory ............................................. 2 9
E rma Z . D robnis
4 B asic Semen Analysis ............................................................................................... 3 9
A shok A garwal, S ajal G upta, and R akesh S harma
5 S emen Analysis Using Hamilton-Thorne Computer Assisted
Semen Analyzer (CASA) ......................................................................................... 4 7
A shok A garwal, S ajal G upta, and R akesh S harma
6 S emen Analysis Using Sperm Class Analyzer (SCA v5) for Computer
Assisted Semen Analysis (CASA) ........................................................................... 5 9
A shok A garwal, S ajal G upta, and R akesh S harma
7 L eukocytospermia Quantitation (ENDTZ) Test ................................................... 6 9
A shok A garwal, S ajal G upta, and R akesh S harma
8 E osin-Nigrosin Staining Procedure ........................................................................ 7 3
A shok A garwal, S ajal G upta, and R akesh S harma
®
9 S perm Morphology Stain (Diff-Quik ) .................................................................. 7 9
A shok A garwal, S ajal G upta, and R akesh S harma
1 0 Q ualitative Seminal Fructose .................................................................................. 8 3
A shok A garwal, S ajal G upta, and R akesh S harma
1 1 C ytospin Procedure and Nuclear Fast Red and Picroindigocarmine
Staining Procedure for Azoospermic Sample ........................................................ 8 5
A shok A garwal, S ajal G upta, and R akesh S harma
1 2 H ypoosmotic Swelling Test (HOS) .......................................................................... 9 3
A shok A garwal, S ajal G upta, and R akesh S harma
1 3 P rocedure for Retrograde Ejaculate ...................................................................... 9 7
A shok A garwal, S ajal G upta, and R akesh S harma
1 4 S perm Preparation for Intrauterine Insemination Using Density
Gradient Separation ................................................................................................ 1 01
A shok A garwal, S ajal G upta, and R akesh S harma
1 5 S perm Preparation for Intrauterine Insemination (IUI)
by Swim-Up Method ................................................................................................ 1 09
A shok A garwal, S ajal G upta, and R akesh S harma
vii
viii Contents
1 6 C ryopreservation of Client Depositor Semen ........................................................ 1 13
A shok A garwal, S ajal G upta, and R akesh S harma
1 7 C ryopreservation of Client Depositor Semen Aspirate ........................................ 1 35
A shok A garwal, S ajal G upta, and R akesh S harma
1 8 C ryopreservation of Client Depositor Testicular Tissue ...................................... 1 39
A shok A garwal, S ajal G upta, and R akesh S harma
1 9 A crosome Reaction Measurement .......................................................................... 1 43
A shok A garwal, S ajal G upta, and R akesh S harma
2 0 D irect SpermMar Antibody Test ............................................................................ 1 47
A shok A garwal, S ajal G upta, and R akesh S harma
2 1 R eactive Oxygen Species (ROS) Measurement ..................................................... 1 55
A shok A garwal, S ajal G upta, and R akesh S harma
2 2 O xidation-Reduction Potential Measurement in Ejaculated Semen Samples ... 1 65
A shok A garwal, S ajal G upta, and R akesh S harma
2 3 A ntioxidant Measurement in Seminal Plasma by TAC Assay ............................. 1 71
A shok A garwal, S ajal G upta, and R akesh S harma
2 4 M easurement of DNA Fragmentation in Spermatozoa
by TUNEL Assay Using Bench Top Flow Cytometer ........................................... 1 81
A shok A garwal, S ajal G upta, and R akesh S harma
2 5 L icensing and Accreditation of the Andrology Laboratory ................................. 2 05
G race M . C entola
I ndex .................................................................................................................................. 2 11
C ontributors
A shok A garwal, P hD, HCLD (ABB), ELD (ACE) A ndrology Center and Reproductive
Tissue Bank, A merican Center for Reproductive Medicine, Cleveland Clinic, C leveland, O H,
U SA
G race M . C entola, P hD, HCLD/CC/ALD (ABB) M acedon, N Y, U SA
E rma Z . D robnis, P hD Department of Obstetrics, Gynecology and Women’s Health,
Reproductive Medicine and Fertility, School of Medicine, U niversity of Missouri, C olumbia,
M O, U SA
S ajal G upta, M D (Ob/Gyn), MSc (Embryology), TS (ABB) A ndrology Center and
Reproductive Tissue Bank, A merican Center for Reproductive Medicine, Cleveland Clinic,
C leveland, O H, U SA
A hmad M ajzoub, M D D epartment of Urology, C leveland Clinic, C leveland, O H, U SA
E dmund S abanegh J r., M D D epartment of Urology, C leveland Clinic, C leveland, O H, U SA
R akesh S harma, P hD A ndrology Center and Reproductive Tissue Bank, American Center
for Reproductive Medicine, C leveland Clinic, C leveland, O H, U SA
ix
