Development of tissue printed nitrocellulose cards/arrays for real time PCR amplification and detection

Abstract

Tissue print technology allows for the transfer of cellular material from tissue onto a nitrocellulose film for immunocytochemical assays. The MIT BioInstrumentation Laboratory is currently developing a novel cancer marker imaging system for detection of cancerous tissue, which will be useful for discerning tumor margins. This research will advance the recent application of tissue print technology in bio-medicine by combining it with imaging and real time polymerase chain reaction (PCR) amplification and detection. A major objective in the design of this instrumentation is to develop the capacity to evaluate much larger areas of tissue. An approach to fulfilling this objective is the creation of a gasket that can seal individual wells of a nitrocellulose array. A gasket was created by laser cutting an assembly of molded silicone rubber and a double-sided tape (silicone-acrylic). Experiments showed when the gasket was adhered to a glass slide and subjected to the PCR, there was no leakage. FAST Slides, nitrocellulose slides provided by Grace Bio-Labs, are cut with a laser to generate the nitrocellulose arrays.