The Advanced Tissue Resources Core (ATRC) provides state-of-the-art molecular pathology resources to the Harvard community. Current resources include Laser Capture Microscopy, Nucleic Acid analysis (Agilent Bioanalyzer), Luminex FlexMap 3D multiplex bead cytometry, and Digital PCR.
Laser Capture Microscopy — One of the consistent characteristics of human disease is that it affects specific tissue regions and cell types. Laser Capture Microscopy (LCM) enables researchers to precisely dissect tissue regions, cell populations or single cells at the microscopic level and then purify nucleic acid and/or protein from these dissectates. Proteins can then be used directly in experiments, while DNA and RNA samples can be expanded through appropriate amplification techniques.
Tissue Quality Analysis — A major problem in the use of banked tissue is the variability of recovery and quality of RNA. Experienced users of the various local brain banks report an approximate yield of less than 50% of the material they need. While the underlying issues that explain this poor yield are complex, it is extremely important to assess nucleic acid quality of a specimen before investing significant time and other resources in subsequent analysis. To provide this 'quality control', the ATRC has an Agilent 2100 Bioanalyzer for unbiased assessment of RNA/DNA quality of tissue samples. miRNA-sized small RNAs can also be visualized with the Agilent bioanalyzer. We have studied the relationship between total RNA stability and miRNA stability using a combination of Agilent bioanalysis, rtPCR and the FlexMiR assay. As such, we are able to reliably assess the quality of miRNA samples prior to expression analyses.
The ATRC operates the latest generation Arcturus Veritas and Arcturus PixCell IIe Laser Capture Microdissection System (with fluorescence illumination), a Luminex FlexMAP 3D system, a Nanodrop 3300 Spectroflourimeter, an Agilent 2100 Bioanalyzer , and a BioRad CFX96 Real Time PCR system and QX200 ddPCR system. The combined power of these instruments allows investigators to capture specific cells from frozen sectioned tissue (including human samples), isolate and amplify cellular RNAs from these captured cells, quickly verify the quality, quantity and fidelity of the RNA obtained, and then perform quantitative expression analyses via qPCR. In addition to LCM services, the ATRC's laboratory supports the protocols for tissue harvesting and banking, frozen sectioning, and immunohistochemistry required prior to LCM. A Thermo-Shandon SME Cryostat and Leica 2155 Motorized Microtome are available for this purpose.
A large part of the ATRC's role is to provide consultation, training, and experimental support for investigators interested in using the facility to its full capacity. We will work with investigators staff on their research assignments and guide their use of the facility.