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Novel flow cytometer techinique for sperm analysis: validation and practical application

Grant number: 14/22972-7
Support type:Scholarships abroad - Research Internship - Master's degree
Effective date (Start): March 01, 2015
Effective date (End): August 31, 2015
Field of knowledge:Agronomical Sciences - Veterinary Medicine - Animal Reproduction
Principal researcher:André Furugen Cesar de Andrade
Grantee:Mariana Andrade Torres
Supervisor abroad: Nestor Gaston Sepulveda Becker
Home Institution: Faculdade de Medicina Veterinária e Zootecnia (FMVZ). Universidade de São Paulo (USP). São Paulo , SP, Brazil
Research place: Universidad de La Frontera (UFRO), Chile  
Associated to the scholarship:13/08070-8 - Effects of seminal plasma on motility characteristics and fertility of cryopreserved boar sperm, BP.MS


This project aims to develop a flow cytometer technique and its applying to analyzer experiment one (master degree project FAPESP n° 2013/08070-8) samples. Fourfold coloration technique will be develop to enable the simultaneous analysis of plasma and acrossome membrane and mitochondrial membrane potential excluding cellular debris. Validation previously, fluorescent probe controls will be realized to clearing the spectral overlap in BD FACSDiva 6.1. Nine ejaculates will be collected do fourfold coloration validation. Semen will be aliquot into five treatments, 100 (100% of in-natura cells), 75 (75% of of in-natura cells plus 25% of fast-frozen cells), 50 (50% of of in-natura cells plus 50% of fast-frozen cells), 25 (25% of of in-natura cells plus 75% of fast-frozen cells) and 0 (100% of fast-frozen cells). (AU)

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Scientific publications
(References retrieved automatically from Web of Science and SciELO through information on FAPESP grants and their corresponding numbers as mentioned in the publications by the authors)
TORRES, MARIANA ANDRADE; DIAZ, ROMMY; BOGUEN, RODRIGO; MASSAMI KITAMURA MARTINS, SIMONE MARIA; RAVAGNANI, GISELE MOURO; LEAL, DIEGO FEITOSA; OLIVEIRA, MELISSA DE LIMA; DONATELLI MURO, BRUNO BRACCO; PARRA, BEATRIZ MARTINS; MEIRELLES, FLAVIO VIEIRA; et al. Novel Flow Cytometry Analyses of Boar Sperm Viability: Can the Addition of Whole Sperm-Rich Fraction Seminal Plasma to Frozen-Thawed Boar Sperm Affect It?. PLoS One, v. 11, n. 8, . (13/08070-8, 14/22972-7, 11/23484-8)

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