Institut Pasteur de Montevideo

2 ARTICLES PUBLISHED IN JoVE

Developmental Biology

Application of the DNA-Specific Stain Methyl Green in the Fluorescent Labeling of Embryos

Daniel Prieto¹, Gonzalo Aparicio^1,2, Matías Machado¹, 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.

Bioengineering

Spatula Montevideo Device for the Vitrification of Mammalian Embryos

Geraldine Schlapp^*1, María Noel Meikle^*1, Lucía Goyeneche², Pedro Dos Santos Neto³, Martina Crispo¹

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.