Biorepository & Tissue Analysis Shared Resource
Chadrick E. Denlinger, MD
Assistant Professor, Department of Surgery
The Hollings Cancer Center (HCC) Biorepository & Tissue Analysis (BTA) Shared Resource provides investigators with access to meticulously collected and annotated human specimens as well as advanced human and animal tissue analyses that require state-of-the-art instrumentation and expertise. The shared resource comprises several integrated components, e.g., biospecimen and data repository, customized and readily available tissue microarrays (TMAs), laser capture microdissection (LCM), matrix-assisted laser desorption/ionization (MALDI) tissue imaging, and research pathology services such as histologic analysis of fixed, frozen, and stained tissues and analysis of experimental results.
The specific objectives of the HCC BTA Shared Resource are to:
- Populate a central biospecimen and data repository.
- Collect, process, and store human tissues according to established standard operating procedures (SOPs) with quality control verified by experienced pathologists at the time of accession.
- Collect, process, and store saliva, urine, plasma, buffy coat, and other protocol-specific human biospecimens at the time of surgery/procedure according to established SOPs.
- Link clinicopathologic data for each human specimen for research use.
- Provide custom human TMAs in a cost-effective manner.
- Provide, develop, and modify tissue analysis services to support cancer research (human and animal), including:
- Routine and custom staining procedures.
- Laser capture microscopy.
- DNA/RNA/protein isolation.
- MALDI tissue imaging and advanced molecular analysis by mass spectrometry.
- Collaborate with other MUSC/HCC entities and shared resources to enhance the capabilities and cost-effectiveness of the BTA.
Equipment and Instrumentation
The BTA Shared Resource has the following items of major equipment:
- Biospecimen Storage: Four -80°C freezers, one -20°C freezer, one 4°C refrigerator, and temperature monitoring systems linked to a central campus monitoring system within MUSC’s Facilities and Engineering Unit.
- Cryostat: Leica CM 1850 UV.
- Microtome: Microm HM 340E.
- Tissue Embedding System: Leica EG 1160.
- Tissue Processor: Milestone Histos 5 Rapid Microwave Histoprocessor.
- Laser Capture Microdissection: Molecular Devices Arcturus XT.
- Bioanalyzer: Agilent 2100.
- MALDI Tissue Imaging: This consists of a dedicated Solarix 70 dual source FTICR MALDI-MS mass spectrometer and AutoFlex III MALDI-TOF/TOF platform. An automated matrix sprayer, nanoliter spotting robot and cryostat also are available at this location.
- Data Storage: The BTA uses TissueMetrix (Artificial Intelligence in Medicine, Inc.) as the biorepository management database. This web-based software utilizes a standard PC workstation with internet connection as well as a Brady printer for label generation. TissueMetrix is hosted locally and data is backed up per institutional protocols. This workstation is physically located within the BTA laboratory and serves at the time when specimens are received along with accompanying clinical data. This workstation provides BTA staff with a laser scanner to easily and efficiently check bar-coded samples out of the facility. This workstation has standard office and graphics programs installed to support various data analyses.
The BTA offers a wide range of services including:
- Consent and Processing for Biospecimen Collection: The BTA maintains IRB approval for a blanket consent that permits the procurement of biospecimens for general banking to be used for future research as well as biospecimens collected specifically for research studies already IRB-approved. The blanket consent also allows patients to continue donating specimens for up to one year from the consent date. The consent allows the specimens collected to be used for genetic research and patients agree to be re-contacted for up to five years from the consent date for follow-up clinical data.
- Clinical Data Linking: When researchers request clinical data elements to accompany specimens, the BTA serves as an honest broker and provides de-identified clinical data to researchers upon receipt of appropriate IRB approval for their research. If the researcher has approval for identified clinical data, the BTA can provide this as well under the BTA’s IRB-approved protocol. Access to clinical and pathologic data is available via MUSC’s enterprise-wide clinical data warehouse that integrates the clinical and research IT systems.
- Tissue Microarrays: As a resource to facilitate high-throughput evaluations, the BTA provides HCC-prepared TMAs for research use. Additional services include assisted TMA construction, complete custom consultation, and construction of TMA blocks. Routine immunohistochemical staining, analysis, and images of results for the TMA slides are also provided.
- Tissue Fixation, Processing, and Embedding: Both animal model and clinical research require the production of high quality tissue samples for morphological and molecular analyses. Quality control of tissue collected by the BTA is performed by a research pathologist, Dr. Shao. This includes verification of disease, evaluation of tissue quality, and assessment of the percent of tumor in each sample. Frozen and FFPE tissues are routinely sectioned. Standard H&E staining and immunohistochemistry staining services are also available.
- Tissue Analysis Services (animal and human): As mentioned earlier, the BTA provides a number of services to support the various needs of HCC members.
- MALDI Tissue Imaging: With specific expertise in MALDI mass spectrometry imaging (MALDI-MSI), the BTA can offer researchers this novel service to spatially profile proteins, lipids, and drug and small molecule metabolites in tissues. Using this approach, a global snapshot of major cellular biomolecules is obtained, which includes their tissue localization and distribution, structure, and relative concentration. The preparation and analysis workflows are readily linked to standard histopathology procedures and the method can be applied to any frozen tissue type. Unique to MUSC and BTA users is the ability to use this method to characterize differences in the expression of glycans, glycolipids, and sphingolipids in tissues of clinical interest. Detection of drug distribution and metabolism in tissues derived from pre-clinical model systems is also available. Analysis of peptides and glycans in formalin-fixed tissues is an additional service.
- RNA/DNA Extraction: Cellular DNA, RNA, and protein prepared by the BTA from banked specimens or any other biomolecules submitted by investigators can be qualitatively assessed by the Agilent Bioanalyzer, prior to use for downstream applications such as microarray and/or qRT-PCR analysis.
- Laser Capture Microdissection: This service enables researchers to identify specific cellular elements under a microscope and isolate them by laser dissection and collections. In this way, highly enriched regions of the tumor or stroma can be evaluated at the molecular level. This service can be performed with human and animal tissue. Collected cells and images of extracted regions are provided.
- Immunohistochemistry Services: The BTA carries out the development of novel immunohistochemistry techniques involving new antibodies and other reagents. This includes the evaluation and quantitation of staining in tumor samples, as well as slide images of results.
Prior to services being performed, the requestor must submit an Order Form. Upon receipt, the operations manager will contact the requestor, review the request, request additional information if needed, and provide a pricing proposal. Please note: all requests for specimens undergo committee review for approval.
- Order Form
- Patient Consent Form
- Leukemia Requisition
- Lymphoma Requisition
- Myeloma Requisition
- Fee Schedule
- Flowsheet to Determine Type of IRB Application
- This tool assists investigators in determining what IRB review category is appropriate for their research.
If you publish a manuscript including research supported by the Biorepository & Tissue Analysis Shared Resource, the HCC recommends the following text to be placed in the acknowledgement section: "Supported in part by the Biorepository & Tissue Analysis Shared Resource, Hollings Cancer Center, Medical University of South Carolina."
If you have any questions please e-mail the Biorepository & Tissue Analysis Shared Resource.