Basic Science

Protocols

  • Complete Follice Guidebook:
    • Follicle Handbook: The handbook describes follicle isolation, encapsulation in alginate, and in vitro culture.  In addition, there are protocols for encapsulation in a fibrin/alginate blend, removal of follicles from alginate, and in vitro maturation of oocytes following in vitro follicle culture. 
    • Reagent and Supply List for Working with Follicles: A comprehensive list detailing everything necessary to set up an in vitro follicle growth system. 
    • Certification for working with follicles: Listed here are the requirements that should be fulfilled prior to starting experiments to ensure that follicle isolation, encapsulation, and culture methods are being performed properly.
  • Follicle Health and Quality Reference Guide: This guide provides useful parameters for assessing the initial quality of isolated follicles and for monitoring follicle health and growth during culture.
  • Whole Follicle Fixation and Staining: This protocol describes how immunocytochemistry can be performed on intact follicles.

Videos

  • Follicle Culture in Alginate Hydrogels. This 9-part video series presented by the Biomaterials Core provides a comprehensive “how to” tour of follicle culture techniques starting from ovary dissection and ending with follicle recovery from alginate following culture. Watch the videos.
  • Human Follicles.  These 3-dimensional movies of confocal reconstructions, taken by Susan Barrett, PhD, highlight the architecture of human follicles. 
    1. In Vitro Grown Human Follicle with Antral Cavity. 3D movie generated from a 100 mm confocal z-stack (Zeiss LSM 510, 40x) of a human, 14-day in vitro grown follicle. This movie shows the newly formed antral cavity, the differentiation of a clear cumulus cell mass around the oocyte, and the mural granulosa cell layers. [F-Actin (red), Nuclei (blue)]. Movies were made on Volocity (Improvision/PerkinElmer). Watch the video.
    2. Transzonal Projections in Immature in Vivo Human Follicles. 3D movie generated from a 100 mm confocal z-stack (Zeiss LSM510, 40x) of a fresh-fixed, immature, human antral follicle showing the transzonal projections between the growing oocyte and the cumulus granulosa. [F-Actin (red), Nuclei (blue)].

Clinical Medicine

Videos