Motility and other characteristics of human sperm can be measured by computer-assisted sperm analysis of samples stained with Hoechst 33342**Supported in part by a subcontract SBIR 2R44 ES0562 to Cornell University, Ithaca, New York, from The Small Business Innovative Research Program, National Institutes of Health, Bethesda, Maryland.††Hoechst 33342, Sigma Chemical Company, St. Louis, Missouri.

1996 ◽  
Vol 66 (3) ◽  
pp. 446-453 ◽  
Author(s):  
Paul B. Farrell ◽  
Robert H. Foote ◽  
Michael J. Zinaman
Keyword(s):  
New York ◽  
Human Sperm ◽  
National Institutes Of Health ◽  
Computer Assisted ◽  
Hoechst 33342 ◽  
Sperm Analysis ◽  
Cornell University ◽  
Chemical Company ◽  
Computer Assisted Sperm Analysis ◽  
Sigma Chemical

Computerized human sperm analysis

1994 ◽  
Vol 61 (1) ◽  
pp. 75-80
Author(s):  
G. Bonanni ◽  
A. Colonna ◽  
S. Masiero ◽  
I. Mastrogiacomo
Keyword(s):  
Human Sperm ◽  
Computer Assisted ◽  
Penetration Test ◽  
Sperm Analysis ◽  
Head Displacement ◽  
Seminal Parameters ◽  
Mucus Penetration ◽  
Computer Assisted Sperm Analysis ◽  
Sperm Motion ◽  
Lateral Head

The recent introduction of various devices for computer-assisted sperm analysis allows us to have not only objective measurements of the seminal parameters but also to determine some characteristics of sperm motion such as curvilinear and linear velocity and amplitude of lateral head displacement that cannot be obtained by routine analysis. These new seminal parameters seem to be very useful in the diagnosis of male infertility and, among them, ALH seems to play a more important role; in one of our studies we found a significant correlation between this parameter and the results of the bovine cervical mucus penetration test (Penetrak). Furthermore it is possible to determine sperm subpopulations for each parameter and so it will be possible to express a more accurate judgement not only on the fertilizing capacity of semen but also to know the effects on spermatozoa of different activating and capacitating treatments.

263 EVALUATION OFALTERNATIVE CRYOPROTECTANTS AND CHOLESTEROL-LOADED CYCLODEXTRINS FOR INCREASING CRYOSURVIVAL OF SEXED REFROZEN STALLION SPERMATOZOA

2008 ◽  
Vol 20 (1) ◽  
pp. 211 ◽  
Author(s):  
J. K. Graham ◽  
M. A. Meyers ◽  
E. L. Squires ◽  
J. L. Schenk
Keyword(s):  
Intracytoplasmic Sperm Injection ◽  
Computer Assisted ◽  
Hoechst 33342 ◽  
Sperm Analysis ◽  
Commercial Use ◽  
Nitrogen Vapor ◽  
Fort Collins ◽  
Viable Sperm ◽  
Computer Assisted Sperm Analysis ◽  
Separation Test

Commercial use of sex-sorted stallion sperm will depend upon the ability to cryopreserve the sperm. In many cases the desired sperm for sorting is already cryopreserved; therefore, this will necessitate the need to thaw, sex-sort, and subsequently refreeze the sperm for future use in intracytoplasmic sperm injection (ICSI). Experiments were conducted to determine if previously frozen stallion spermcould be thawed, sex-sorted, and cryopreserved. Ejaculates from 6 stallions were frozen in FR5 cryopreservation medium at 400 million sperm mL–1 (0.5-mL straws). Samples were thawed for 30 s in 37�C water and diluted (100 million sperm mL–1) in Kenny's modified Tyrode's medium (KMT). Samples (2 mL) were stained with 36.53 µm Hoechst 33342 in 34�C water for 45 min and then diluted to 75 million sperm mL–1 with KMT containing 0.002% food coloring dye (FD&C #40). Sperm were sorted using an SX MoFlo™ (Dako, Fort Collins, CO, USA), and the sorted samples centrifuged at 850g for 10 min. The sperm were suspended to 4.6 million sperm mL–1 in FR5 + 0.54 m glycerol (control) or the following treatments: FR5 + 0.52 m dimethylformamide (DMF); FR5 + 0.54 m glycerol + 1.5 mg cholesterol-loaded cyclodextrins (CLC)/120 million cells; FR5 + 0.52 m DMF + 1.5 mg CLC/120 million cells. Non-sorted control sperm (NSC) were removed from original frozen/thawed samples and suspended to 4.6 million sperm mL–1 in FR5 + 0.52 m DMF. Sorted and non-sorted samples were cooled from 22�C to 4�C over 2 h, loaded into 0.25-mL straws, and frozen in liquid nitrogen vapor. Straws were thawed in 37�C water for 30 s and the percentages of motile and viable sperm were determined using computer-assisted sperm analysis and flow cytometry (5 million cells were stained with 10 µL propidium iodide and 20 µL SYBR-14), respectively. Data were analyzed by analysis of variance and treatment means were separated using the Student-Newman-Keuls mean separation test. The percentages of total motile spermatozoa were higher for frozen/thawed sexed refrozen (FTR) sperm refrozen in DMF + CLC (10%) than for the glycerol control (2%; SEM � 1; P < 0.05), but this motility was similar to that of the DMF control (5%), the glycerol + CLC treatment (4%), and the NSC (7%; SEM � 1; P > 0.05). The percentages of viable spermatozoawere higher for FTR (glycerol and DMF) sperm treated with CLC prior to freezing (68% and 71%) than for control sperm (58% and 56%; SEM � 3; P < 0.05), respectively. Frozen/thawed sexed refrozen samples resulted in significantly higher percentages of viable spermatozoa compared to NSC (42%; SEM � 3; P < 0.05). These findings indicate that cryopreserved stallion sperm can be thawed, sex-sorted, and refrozen and still maintain a high percentage of viable spermatozoa; treating sperm with CLC prior to refreezing improved cryosurvival for both cryoprotectants. This may provide adequate numbers of viable sperm for use in intracytoplasmic sperm injection procedures. This work was funded by XY, Inc.

scholarly journals Cyclic FEE Peptide Improves Human Sperm Movement Parameters without Modification of Their Energy Metabolism

2021 ◽  
Vol 22 (20) ◽  
pp. 11263
Author(s):  
Nathalie Le Foll ◽  
Jean-Christophe Pont ◽  
Audrey L’Hostis ◽  
Thomas Guilbert ◽  
Frédéric Bouillaud ◽  
...  
Keyword(s):  
Glutamic Acid ◽  
Human Sperm ◽  
Control Group ◽  
Computer Assisted ◽  
Atp Production ◽  
Sperm Analysis ◽  
Mitochondrial Functions ◽  
Computer Assisted Sperm Analysis ◽  
Movement Parameters ◽  
Sperm Movement

Cyclic fertilin peptide (cFEE: phenylalanine, glutamic acid; glutamic acid) improves gamete interaction in humans. We investigate whether it could be via improvement of sperm movement parameters and their mitochondrial ATP production. Sperm movement parameters were studied using computer-assisted sperm analysis (CASA) in sperm samples from 38 patients with normal sperm in medium supplemented with cyclic fertilin against a control group. Sperm mitochondrial functions were studied using donor’s sperm, incubated or not with cFEE. It was evaluated by the measurement of their ATP production using bioluminescence, their respiration by high resolution oxygraphy, and of mitochondrial membrane potential (MMP) using potentiometric dyes and flow cytometry. cFEE significantly improved sperm movement parameters and percentage of hyperactivated sperm. Impact of inhibitors showed OXPHOS as the predominant energy source for sperm movement. However, cFEE had no significant impact on any of the analyzed mitochondrial bioenergetic parameters, suggesting that it could act via a more efficient use of its energy resources.

Сравнительная оценка спермопродукции чистопородных и гибридных хряков-производителей

2017 ◽  
pp. 2-6
Author(s):  
Байлар Садраддинович Иолчиев ◽  
Анна Валиевна Таджиева ◽  
Павел Михайлович Кленовицкий ◽  
Вугар Алиевич Багиров ◽  
Александр Алексеевич Никишов ◽  
...  
Keyword(s):  
Computer Assisted ◽  
Sperm Analysis ◽  
Computer Assisted Sperm Analysis

Практической частью репродуктивной технологии является прогноз и оценка фертильности самцов. Существующие методы оценки семени не дают достаточно корректного прогноза результатов осеменения и, следовательно, репродуктивного потенциала того или иного производителя. В работе приведены данные комплексной оценки биологической полноценности семени чистопородных и гибридных хряков-производителей с использованием компьютерной технологии CASA (computer-assisted sperm analysis). Оценивали активность сперматозоидов в образцах классическим способом (микроскопирование) и с помощью компьютерной технологии, программа «Зоосперм 1.0». Исследования проводили в 2014 – 2015 гг. в Лаборатории репродуктивной криобиологии животных (Россия). Объектом исследования служила сперма чистопородных и гибридных хряков-производителей, разводимых в разных регионах Российской Федерации: крупная белая (n = 24), дюрок (n = 28), ландрас (n = 18), пьетрен (n = 8), гибридные (n = 6). Установлено, что высокими репродуктивными качествами отличаются хряки-производители породы ландрас и гибридов, от них в среднем за эякулят получено 290,2 и 290,6 мл спермы соответственно; наименьшими — хряки-производители породы пьетрен (270 мл). Частота встречаемости сперматозоидов с аномальной морфологией высокая у хряков-производителей породы дюрок (12 %) и пьетрен (15 %). Частота встречаемости аномалии в разных сегментах сперматозоидов зависит от индивидуальной особенности хряков-производителей. У одних производителей наиболее часто встречаются морфологические отклонения головки, например, у хряка породы ландрас оно составило 58,60 %, у других часто встречались отклонения от нормы в строении жгутика (12,1 – 29,3 %). Использование компьютерной технологии при оценке биологической полноценности сперматозоидов хряков-производителей является наиболее точным и информативным.

scholarly journals Post-Thaw Sperm Quality and Functionality in the Autochthonous Pig Breed Gochu Asturcelta

Animals ◽  
2021 ◽  
Vol 11 (7) ◽  
pp. 1885
Author(s):  
José Néstor Caamaño ◽  
Carolina Tamargo ◽  
Inmaculada Parrilla ◽  
Felipe Martínez-Pastor ◽  
Lorena Padilla ◽  
...  
Keyword(s):  
Genetic Resource ◽  
Sperm Quality ◽  
Genetic Material ◽  
Mitochondrial Activity ◽  
Computer Assisted ◽  
Kinematic Parameters ◽  
Linear Mixed Effects ◽  
Sperm Analysis ◽  
Computer Assisted Sperm Analysis ◽  
Analysis System

Genetic resource banks (GRB) preserve the genetic material of endangered, valuable individuals or genetically relevant breeds. Semen cryopreservation is a crucial technique to reach these goals. Thus, we aimed to assess the sperm parameters of semen doses from the native pig breed Gochu Asturcelta stored at the GRB of Principado de Asturias (GRB-PA, Gijón, Spain), focusing on intrinsic and extrinsic (boar, season) factors. Two straws per boar (n = 18, 8–71 months of age) were thawed, pooled, and assessed after 30 and 150 min at 37 °C by CASA (computer-assisted sperm analysis system; motility and kinematic parameters) and flow cytometry (viability, acrosomal status, mitochondrial activity, apoptosis, reactive oxygen species, and chromatin status). The effects of age, incubation, and season on post-thawing quality were determined using linear mixed-effects models. Parameters were on the range for commercial boar breeds, with chromatin status (SCSA: fragmentation and immaturity) being excellent. Incubation decreased sperm quality and functionality. The boar age did not have a significant effect (p > 0.05), but the between-boar variability was significant (p < 0.001). The season significantly affected many parameters (motility, kinematics, viability, acrosomal status, mitochondrial activity), especially after 150 min of incubation. In general, samples collected in spring and summer showed higher quality post-thawing, the lowest in winter. In conclusion, the sperm doses from the Gochu Asturcelta breed stored at the GRB-PA showed excellent chromatin status and acceptable characteristics after thawing. Therefore, boar and seasonal variability in this autochthonous breed could be relevant for cryobank management.

scholarly journals Evaluación de la motilidad espermática a través del sistema C.A.S.A de semen caprino criopreservado bajo diferentes medios diluyentes

Respuestas ◽  
2013 ◽  
Vol 18 (2) ◽  
pp. 16-27
Author(s):  
Leonardo Hernández-Corredor ◽  
Alexander Nivia-Osuna ◽  
Daniel Hernández-Villamizar ◽  
Jorge Alexander Rubio-Parada ◽  
Armando Quintero-Moreno
Keyword(s):  
Lateral Displacement ◽  
Egg Yolk ◽  
Computer Assisted ◽  
Complete Medium ◽  
Sperm Analysis ◽  
Straight Line ◽  
Nitrogen Vapor ◽  
Computer Assisted Sperm Analysis ◽  
The University ◽  
Casa System

 El estudio evaluó la motilidad espermática y su efecto postdescongelación en semen caprino, en dos medios comerciales (Andromed® y TwoStep®) y diferentes protocolos de congelación (medio completo, con adicción del 10% de yema de huevo, semen centrifugado y sobrenadante seminal), se utilizaron machos de la raza alpina de la Universidad Francisco de Paula Santander Ocaña, el semen fue colectado con electroeyaculador, una vez los medios terminados y parte de los contenidos seminales enteros o centrifugados mezclados, se estabilizó por 2 horas, se envasó en pajillas de 0,5 cc y se congela en vapores de nitrógeno por 10 minutos, las pajillas se llevaron al laboratorio de Andrología de la Universidad del Zulia y por medio del sistema C.A.S.A.(Computer Assisted Sperm Análisis) se evaluaron los parámetros de motilidad como velocidad curvilínea (VCL), velocidad rectilínea (VSL), velocidad lineal (VAP), índice de linealidad (LIN), índice de rectitud (STR), índice de oscilación (ALH), Amplitud media del desplazamiento lateral de la cabeza del espermatozoide (BCF), los datos fueron analizados por medio del procedimiento GLM de SAS versión 9.0; los mejores índices de motilidad (VCL, ALH, BCF) fueron expresados enel tratamiento de contenido seminal centrifugado en medio Andromed®. (p≤0,001))La mejor progresividad espermática (VSL,LIN,STR)se presentó el tratamiento de Semen completo de caprino, criopreservado en medio comercial TwoStep®. ABSTRACT  The study evaluated the effect sperm motility and sperm post-thawing in goats, two commercial means (Andromed ® and Two Step ®) and different freezing protocols (complete medium with 10% addition of the egg yolk, semen centrifuged supernatant and seminal ), we used males of the Alpine race of the University Francisco de Paula Santander Ocaña, semen was collected with electroejaculator once finished media and part of the whole and centrifuged seminal contents mixed, stabilized by two hours, packed in 0.5 cc straws and frozen in nitrogen vapor for 10 min, the straws were taken to the laboratory of Andrology at the University of Zulia and through CASA system (Computer Assisted Sperm Analysis) were evaluated motility parameters such as curvilinear velocity (VCL), straight line velocity (VSL), linear velocity (VAP), linearity index (LIN), straightness index (STR) Oscillation Index (ALH ) average amplitude of the lateral displacement of the sperm head (BCF), the data were analyzed by the GLM procedure of SAS version 9.0, the highest rates of motility (VCL, ALH, BCF) were expressed in the treatment of seminal content centrifugation Andromed ® medium. (p ≤ 0.001)) The best progressive sperm (VSL, LIN, STR) will present the full Semen treatment goats, cryopreserved at Two Step ® commercial medium.Keywords: semen, buck, Andromed, Two step.

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