Table Of ContentAshok Agarwal · Sajal Gupta
Rakesh Sharma Editors
Andrological
Evaluation
of Male Infertility
A Laboratory Guide
123
Andrological Evaluation of Male Infertility
Ashok Agarwal (cid:129) S ajal Gupta (cid:129) Rakesh Sharma
Editors
Andrological Evaluation
of Male Infertility
A Laboratory Guide
Editors
Ashok Agarwal, PhD, HCLD (ABB), ELD (ACE) Sajal Gupta, MD (Ob/Gyn), MS (Embryology),
Andrology Center and Reproductive Tissue Bank TS (ABB)
American Center for Reproductive Medicine Andrology Center and Reproductive Tissue Bank
Cleveland Clinic American Center for Reproductive Medicine
Cleveland , OH , USA Cleveland Clinic
Cleveland , OH , USA
Rakesh Sharma, PhD
Andrology Center and Reproductive Tissue Bank
American Center for Reproductive Medicine
Cleveland Clinic
Cleveland , OH , USA
ISBN 978-3-319-26795-1 ISBN 978-3-319-26797-5 (eBook)
DOI 10.1007/978-3-319-26797-5
Library of Congress Control Number: 2016941225
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Foreword
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!
September 25, 2015 Edmund Sabanegh Jr., MD
Department of Urology
The Cleveland Clinic
Cleveland, OH, USA
v
Contents
1 Diagnostic Tests in the Evaluation of Male Infertility .......................................... 1
Ahmad Majzoub and Edmund Sabanegh Jr.
2 Quality Management in Andrology Laboratory ................................................... 11
Erma Z. Drobnis
3 Competency Assessment in Andrology Laboratory ............................................. 29
Erma Z. Drobnis
4 Basic Semen Analysis ............................................................................................... 39
Ashok Agarwal , Sajal Gupta , and Rakesh Sharma
5 Semen Analysis Using Hamilton-Thorne Computer Assisted
Semen Analyzer (CASA) ......................................................................................... 47
Ashok Agarwal , Sajal Gupta , and Rakesh Sharma
6 Semen Analysis Using Sperm Class Analyzer (SCA v5) for Computer
Assisted Semen Analysis (CASA) ........................................................................... 59
Ashok Agarwal , Sajal Gupta , and Rakesh Sharma
7 Leukocytospermia Quantitation (ENDTZ) Test ................................................... 69
Ashok Agarwal , Sajal Gupta , and Rakesh Sharma
8 Eosin-Nigrosin Staining Procedure ........................................................................ 73
Ashok Agarwal , Sajal Gupta , and Rakesh Sharma
9 Sperm Morphology Stain (Diff-Quik®) .................................................................. 79
Ashok Agarwal , Sajal Gupta , and Rakesh Sharma
10 Qualitative Seminal Fructose .................................................................................. 83
Ashok Agarwal , Sajal Gupta , and Rakesh Sharma
11 Cytospin Procedure and Nuclear Fast Red and Picroindigocarmine
Staining Procedure for Azoospermic Sample ........................................................ 85
Ashok Agarwal , Sajal Gupta , and Rakesh Sharma
12 Hypoosmotic Swelling Test (HOS) .......................................................................... 93
Ashok Agarwal , Sajal Gupta , and Rakesh Sharma
13 Procedure for Retrograde Ejaculate ...................................................................... 97
Ashok Agarwal , Sajal Gupta , and Rakesh Sharma
14 Sperm Preparation for Intrauterine Insemination Using Density
Gradient Separation ................................................................................................ 101
Ashok Agarwal , Sajal Gupta , and Rakesh Sharma
15 Sperm Preparation for Intrauterine Insemination (IUI)
by Swim-Up Method ................................................................................................ 109
Ashok Agarwal , Sajal Gupta , and Rakesh Sharma
vii
viii Contents
16 Cryopreservation of Client Depositor Semen ........................................................ 113
Ashok Agarwal , Sajal Gupta , and Rakesh Sharma
17 Cryopreservation of Client Depositor Semen Aspirate ........................................ 135
Ashok Agarwal , Sajal Gupta , and Rakesh Sharma
18 Cryopreservation of Client Depositor Testicular Tissue ...................................... 139
Ashok Agarwal , Sajal Gupta , and Rakesh Sharma
19 Acrosome Reaction Measurement .......................................................................... 143
Ashok Agarwal , Sajal Gupta , and Rakesh Sharma
20 Direct SpermMar Antibody Test ............................................................................ 147
Ashok Agarwal , Sajal Gupta , and Rakesh Sharma
21 Reactive Oxygen Species (ROS) Measurement ..................................................... 155
Ashok Agarwal , Sajal Gupta , and Rakesh Sharma
22 Oxidation-Reduction Potential Measurement in Ejaculated Semen Samples ... 165
Ashok Agarwal , Sajal Gupta , and Rakesh Sharma
23 Antioxidant Measurement in Seminal Plasma by TAC Assay ............................. 171
Ashok Agarwal , Sajal Gupta , and Rakesh Sharma
24 Measurement of DNA Fragmentation in Spermatozoa
by TUNEL Assay Using Bench Top Flow Cytometer ........................................... 181
Ashok Agarwal , Sajal Gupta , and Rakesh Sharma
25 Licensing and Accreditation of the Andrology Laboratory ................................. 205
Grace M. Centola
Index .................................................................................................................................. 211
Contributors
Ashok Agarwal , PhD, HCLD (ABB), ELD (ACE) Andrology Center and Reproductive
Tissue Bank, A merican Center for Reproductive Medicine, Cleveland Clinic, C leveland, O H,
USA
Grace M. Centola , PhD, HCLD/CC/ALD (ABB) Macedon , NY , USA
Erma Z. Drobnis , PhD Department of Obstetrics, Gynecology and Women’s Health,
Reproductive Medicine and Fertility, School of Medicine, U niversity of Missouri, C olumbia,
MO , USA
Sajal Gupta , MD (Ob/Gyn), MSc (Embryology), TS (ABB) Andrology Center and
Reproductive Tissue Bank, A merican Center for Reproductive Medicine, Cleveland Clinic,
Cleveland , OH , USA
Ahmad Majzoub , MD Department of Urology , Cleveland Clinic , Cleveland , OH , USA
Edmund Sabanegh Jr. , MD Department of Urology , Cleveland Clinic , Cleveland , OH , USA
Rakesh Sharma , PhD A ndrology Center and Reproductive Tissue Bank, American Center
for Reproductive Medicine , Cleveland Clinic , Cleveland , OH , USA
ix
