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Advanced Tissue Resources Core

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Thanks to a major new grant, we are pleased to announce that as of November 2012 the Advanced Tissue Resources Core will be open to investigators from beyond the Harvard community. Please refer to our revised “Access” guidelines below for further details. 


 

Background

The Advanced Tissue Resources Core (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. 

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

Although the Advanced Tissue Resources Core primarily serves the Harvard community, including the 17 affiliated research hospitals, starting November 2012 we will also welcome users from other parts of the Massachusetts academic neurobiology research community. 



When demand outstrips supply, we will prioritize access based on a combination of the relevance of the projects to the neurobiological, neurological and neuropsychiatric mission of the Center, and the source of the funds supporting the core. 



In practice this is likely to mean that whereas the bulk of our users will continue to be from the wider Harvard-affiliate community, there will also be considerable access for non-Harvard, Massachusetts investigators. 



The ATRC is a fee-for-use facility that provides state-of-the-art molecular pathology resources and training. Investigators may request work to be done by the core staff, or may arrange to use the facility’s equipment themselves.

User fees vary 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.

Before using the facility, investigators are required to submit an ATRC user form including a brief research summary of your proposed project. At this point you are also encouraged to visit or otherwise contact the ATRC to discuss the proposed work. For more information, or to schedule an initial visit, please contact the ATRC. Director, Dr. This e-mail address is being protected from spam bots, you need JavaScript enabled to view it .

After submitting a user form, you should use the ATRC online calendar to check instrument availability prior to booking time. All booking is done directly with ATRC staff. 



If you intend to use the facility’s equipment yourself, the director will determine if you first need training. For NeuroDiscovery members, the first 5 hours of training/consultation/instrument use is free. This initial period is intended to provide a first time user with preliminary data/proof of concept regarding their project. Thereafter, and for non-HNDC members, the standard fees apply. 



Although priority is given to academic neuroscience investigators, the facility is also available to other academic investigators within the greater-Boston research community. And under special arrangements the facility may also be made available to the commercial sector. Please contact ATRC Director, Dr. Charles Vanderburg, for details.