Microfabricated Enumeration Grid Chambers (MEGC)
MEGAN CHESNUT - LSU BIOLOGICAL ENGINEERING
Cryopreservation is an essential tool to bank valuable genetic resources. However, a lack of standardization and reproducibility impedes the development of germplasm repositories. The goal of this work was to evaluate the feasibility of microfabrication techniques to produce low-cost disposable counting chambers for standardized estimation of zebrafish sperm concentration. These microfabricated enumeration grid chambers (MEGC) (Fig. 1) were composed of a polydimethylsiloxane (PDMS) coverslip with grid patterns (100 × 100 μm) and a PDMS base platform (Fig. 2) to create a known volume with a 10-μm height to restrict sperm cells to a single layer. The results of cell counts (Fig. 3) estimated by two of three MEGC devices tested were not significantly different from a commercially available Makler® chamber. The material cost for a MEGC was < $0.1 compared to ~ $100 for a standard hemocytometer and $700 for a Makler® counting chamber. Although fabrication quality could be further improved, this study demonstrates the feasibility of soft lithography methods in creating low-cost, standardized counting chambers for the zebrafish research community to enhance development of comprehensive germplasm repositories.