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Accurate determination of sperm concentration in aquatic species is important for assisted reproduction and cryopreservation, yet is challenging as current counting methods are costly or not suitable for many species. The goal of this work was to develop a simple (single-piece and single-layer photolithography) sperm counting chamber (SSCC) for aquatic species. Goldfish ( Carassius auratus) and zebrafish ( Danio rerio ) sperm were used for evaluation in the device, which was created with soft lithography. Four designs with different geometries were evaluated for counting accuracy. Open-corner and open-midpoint designs were the most accurate with no significant differences ( P &gt; 0.05) for most of the target sperm concentrations (0.5-1.0 × 10 8 cells/mL). The open-corner design was not significantly different from the Makler ® counting chamber intended for human sperm cells ( P = 0.6) but was significantly different from a hemocytometer ( P &lt; 0.001) intended for other cell sizes. Material cost of device production was USD 16 per unit, including photolithography supplies, glass slide and coverslip, and polydimethylsiloxane. The cost can be reduced to USD 2 per unit with repeated wafer casts. This device could be further refined for resin 3-D printing and sharing via open-hardware approaches and modified to best suit species specific applications.

Development of a Single-Piece Sperm Counting Chamber (SSCC) for Aquatic Species