A filtration medium replacement method that increases the efficiency of immunofluorescence staining of oocytes

Immunofluorescence staining is used to investigate proteins and protein interactions in oocytes. In typical protocols, the medium that suspends the oocytes requires replacement more than ten times during the staining procedure; this is time-consuming, technically challenging and not amenable to automation. We developed a filtration method using negative pressure to replace manual replacement of medium. We investigated oocyte loss, time required and staining results using our filtration method compared to traditional procedure. We found that our filtration method reduced oocyte loss by at least 60% and decreased the time required to obtain comparable staining outcomes. It provides an efficient and fast way to replace culture medium for oocytes.