Testing and Validation of an Autostainer

Histology performed by Cal Poly students for research projects and labs are currently performed by hand using a dipping technique. This technique is time consuming, expensive, imprecise, and often unrepeatable. Central Coast Pathology donated a Dako Autostainer to the Biomedical Engineering Department that can perform immunohistological stains of up to 48 samples simultaneously using tightly controlled reagent volumes and incubation times. Protocols are programmed into the machine and can be repeated under the same conditions every time with minimal preparation and exposure to potentially dangerous reagents. The autostainer was repaired and validated by comparing PECAM & BBI stains performed on human umbilical vascular endothelial cells (HUVEC) with the autostainer. HUVEC cells contain the extracellular antigen that PECAM binds to and DNA which BBI interacts with. The stain procedure was a success as the images of the stained HUVEC contained fluorescent portions from both PECAM and BBI. Finally a guide was developed to assist students to successfully create and run their own protocols using the now functional autostainer