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Institut Pasteur de Montevideo

2 ARTICLES PUBLISHED IN JoVE

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Developmental Biology

Application of the DNA-Specific Stain Methyl Green in the Fluorescent Labeling of Embryos
Daniel Prieto 1, Gonzalo Aparicio 1,2, Matías Machado 1, Flavio R. Zolessi 1,2
1Institut Pasteur de Montevideo, 2Sección Biología Celular, Facultad de Ciencias, Universidad de la República

A method for fluorescent staining of fixed biological material with the specific DNA label methyl green is described. Methyl green is used in a diluted aqueous solution and is very resistant to photobleaching. Its far-red emission allows for deep specimen imaging, making it particularly adequate for whole embryos.

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Bioengineering

Spatula Montevideo Device for the Vitrification of Mammalian Embryos
Geraldine Schlapp *1, María Noel Meikle *1, Lucía Goyeneche 2, Pedro Dos Santos Neto 3, Martina Crispo 1
1Laboratory Animal Biotechnology Unit, Institut Pasteur de Montevideo, 2Centro de Esterilidad Montevideo, 3Fundación IRAUy, Instituto de Reproducción Animal de Uruguay

Here, we present the Spatula Montevideo protocol for vitrifying mouse and sheep pre-implantation embryos, consisting of a homemade spatula and utilizing homemade solutions. Embryos are rapidly equilibrated in two cryoprotectant solutions of increasing concentration, loaded onto the spatula, immersed in liquid nitrogen, and finally sealed with a 0.5 mL straw.

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