Advanced Tissue Resource Center

The NeuroDiscovery Advanced Tissue Resource Center (ATRC) provides state-of-the-art molecular pathology resources to the Harvard community. Current resources include laser capture microscopy, DNA/RNA/miRNA quality/expression analysis, Luminex FlexMap 3D multiplex bead cytometry, and real-time PCR.

miRNA Expression Analysis — We have recently built upon the ATRC’s expertise by creating a miRNA core facility that allows investigators to examine the central nervous system miRNA profile and determine the relevance of miRNA species to AD and other neurodegenerative diseases. Leading edge technology and expert scientific support dedicated to the neurodegenerative diseases provides ATRC investigators with ready access to this emerging and increasingly relevant area of research.  miRNA services include tissue and LCM-derived miRNA collections and amplifications, and Human/Rodent miRNA expression analyses based on the Luminex FlexMAP 3D/FlexMiR miRNA assay.

Laser Capture Microscopy — One of the consistent characteristics of neurodegenerative diseases is that they affect specific and stereotyped parts of the nervous system. Laser Capture Microscopy (LCM) enables researchers to precisely isolate material from these regions at the microscopic level. Investigators are able to isolate DNA, mtDNA, mRNA, miRNA and protein from precise populations of neurons. Proteins can then be used directly in experiments, while DNA and RNA samples can be expanded through appropriate amplification techniques.

DNA/miRNA 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 biologic issues that explain this poor yield are complicated, 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.
 

Program Resources

Equipment

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 iCycler Real Time PCR 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.

Reagents for Neurological Research

As part of the Harvard NeuroDiscovery Center's commitment to furthering research within the Harvard neuroscience community, the ATRC makes certain reagents available to NeuroDiscovery members.  The ATRC Reagent Bank contains a number of general utility bio-reagents that we have designed, produced and/or characterized in our laboratory for neuroscience research. Included is the ATRC Primer Panel which we use in conjunction with a set of Brain cDNAs in our BioRad qPCR assay. 

The ATRC reagent bank sometimes contains surplus tissue-derived reagents such as Brain RNAs, LCM dissected samples, and cryosections. The ATRC also hopes to garner new additions to our reagent bank coincident with projects carried out by NeuroDiscovery members. Collaboration-minded investigators interested in sharing research reagents that they have generated in their own laboratories are invited to contribute to our Reagent Bank. Please send a message containing germane details and your contact information to ATRC Director Dr. This e-mail address is being protected from spam bots, you need JavaScript enabled to view it , and we will post it along with the appropriate recognition.

Training

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.  This e-mail address is being protected from spam bots, you need JavaScript enabled to view it and Meredith Banigan are on hand to work with investigators on their latest research and to guide investigators in their use of the facility.

ATRC Online Library

A list of research publications and laboratory protocols relevant to the needs and interests of our collaborators has been compiled. Please visit the ATRC’s online library to download these documents in PDF format.

Access

The ATRC is a fee-for-use facility. For NeuroDiscovery members, the first 10 hours of training, consultation and instrument use is free. This initial period is intended to provide a first time user with preliminary data/proof of concept regarding their project, and is generally sufficient when investigators work under the aegis of ATRC staff. Thereafter, the base fee varies depending on activity (see complete list of charges ) and the extent of your proposed work. Project-based charge-back agreements for large projects can be negotiated with the ATRC Director, Dr. Charles Vanderburg.

Before using the facility, investigators are required to submit an ATRC user form including a brief research summary of their proposed project to the core director. For more information, or to schedule your initial visit, please contact the ATRC.

Users should use the ATRC online calendar to check instrument availability prior to booking time. All booking is done directly with ATRC staff.

Although priority is given to NeuroDiscovery members investigating neurodegenerative diseases and the CNS, the facility is also available to any academic investigators within the Harvard medical community and the greater-Boston research community. Under special arrangements the facility may also be made available to the commercial sector. Please contact ATRC Director, Dr. Charles Vanderburg, for details.