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11 A Novel Multi-Well Panel Staining System: Ideal for the standardization of Immunohistochemistry (IHC), In-Situ Hybridization (ISH) and Fluorescence In-Situ Hybridization (FISH).

Background: IHC, ISH and FISH have played an important role in diagnostic clinical and research pathology. An increasing number of these assays are performed on glass slide tissue. In recent years, automated staining systems have been successfully adopted in large clinical laboratories. However in small and medium size laboratories, the use of expensive automated equipment may be prohibitive

Design: In this study, we designed a novel slide-holder with multiple wells to perform up to 3-10 simultaneous immunohistochemical, ISH, or FISH assays. A variety of antibodies, DNA probes and reagents were coated and dried on the multi-well trays. IHC, ISH and FISH were performed between the slides (attached to a holder) and the coated wells. Routine methods of IHC, ISH and FISH performed on the same tissue were used as comparison controls. The antibodies included CD20, CD3, CD30, CD15 and CD5. EBER and EBV DNA were used for ISH; WCP7, WCP10 and WCP13 were used for FISH. Specific antigen or RNA/DNA controls (corresponding to the coated antibody or probe) were positioned on the slides (built-in) above the test tissue. Consistency was checked by using serial tissue sections assayed with the same antibody or probe. Results: There was no difference in positivity, intensity or background using IHC (10 cases), ISH (4 cases) and FISH (3 cases) with the multi-well technique compared to routine methodology. Built-in antigens, RNA or DNA were adequate for both positive and negative controls and confirmed that the correct antibody or probe was used on the test slide. Using the multi-well method, productivity was increased by 240-300 slides/person/day (300%). Turn around time was vastly reduced making it possible with frozen tissue to complete IHC in 15 minutes or FISH in 10 minutes (quick FISH) if necessary.

Conclusions: The slide-holder and reagent coated multi-well system provides a controlled delivery system, which does not require a computer or robotics. Built-in controls or labels coated on the test slides offer quality control and a reaction indicator. This simple system is a convenient, efficient and economic alternative to high cost automated systems and therefore, ideal for small and medium volume laboratories.

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