An easy, reproducible and cost-effective method for andrologists to improve the laboratory diagnosis of non-obstructive azoospermia: a novel microcentrifugation technique

Abstract

This study describes a new method of microcentrifugation as an improved, viable, cost-effective option to the classical Cytospin apparatus to confirm azoospermia. Azoospermic semen samples were evaluated for cryptozoospermia by a centrifugation method similar to that of World Health Organization guidelines (2010; entire specimen centrifuged at 3000g for 15 min, and aliquots of the pellet examined). Then, if no sperm were detected, the pellet from that procedure was resuspended in culture medium, centrifuged (2000g for 15 min), and the entire pellet spread on a 4 X 6mm area of a slide and stained using the Christmas tree method (Nuclear-Fast solution and picric acid). The entire stained area was examined for the presence or absence of sperm. A total of 148 azoospermic samples (after standard WHO diagnosis) were included in the study and 21 samples (14.2%) were identified as sperm-positive. In all microcentrifugation slides, intact spermatozoa could be easily visualized against a clear background, with no cellular debris. This novel microcentrifugation technique is clearly a simple and effective method, with lower cost, increasing both sensitivity and specificity in confirming the absence or presence of spermatozoa in the ejaculate. It may represent a step forward of prognostic value to be introduced by andrology laboratories in the routine evaluation of patients with azoospermia in the initial semen analysis.

Tissue and sperm handling before assisted reproductive technology (ART): A systematic review

“…A positive correlation was observed between both analyses, concluding that the use of staining may avoid a surgical testicular biopsy procedure. NF-PICS can be easily included in andrology laboratories routines as an inexpensive and effective way to identify sperm in azoospermic patients [ 33 , 34 ].…”

Novel concepts in male factor infertility: clinical and laboratory perspectives

“…Ejaculates of men with NOA usually have normal volume and pH, which indicates patent ejaculatory ducts and functional seminal vesicles [1]. It is important that such specimens are examined after high-speed (>1000g) centrifugation to exclude the presence of rare spermatozoa [70]. The finding of any spermatozoa may offer the chance of ART to be performed with ICSI intracytoplasmic sperm injection, micro-TESE microdissection testicular sperm extraction, NOA nonobstructive azoospermia, OA obstructive azoospermia, PCR polymerase chain reaction, SR sperm retrieval, T/E testosterone to estradiol ratio (T and E units of measurement: ng/dL and pg/ mL, respectively), TESE testicular sperm extraction, TT total testosterone ejaculated sperm, thus avoiding sperm retrieval (SR) methods.…”

Identification of PDCL2 as a candidate marker in Sertoli cell-only syndrome by chromatin immunoprecipitation-sequencing and bioinformatics analysis

“…Clinically, men with NOA and SCOS are likely to present with testicular hypotrophy or even atrophy and abnormal sexual hormones. It is crucial that men obtain at least two semen analyses and proceed to centrifugation of semen samples before initial diagnosis is established ( 15 ). Karyotyping and YCMD testing should be routinely performed to assess potential risks to the offspring ( 16 ).…”