Molecular Histopathology and Analytical Microscopy Core (MHAM)


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The Molecular Histopathology and Analytical Microscopy Core (MHAM) is a new scientific core that works in concert with the CIMC and TGC Cores. The primary mission of the MHAM is to provide advanced detection, imaging, and analysis of gene and protein dynamics in cellular models of normal and cancer tissues from research animals or from patients. The MHAM has state-of-the-art histochemical and immunohistochemical methodologies to determine protein expression, localization, and interactions between viral and cellular proteins. Molecular histopathology, including in situ hybridization, is used to provide investigators with information regarding cellular and viral DNA and/or RNA. Given the translational nature of the COBRE and its  role in the LSU Cancer Center, this core is led by a trained pathologist who, in addition to imaging services, also provides histopathological information on patient tissues and provides access to clinical samples that are crucial to investigate the molecular and cellular pathways correlating with human disease. Finally, the MHAM core provides investigators with training on tissue processing and management and helps investigators develop the necessary imaging and data for the submission of manuscripts and grants.

FACILITIES AND OTHER RESOURCES

INSTITUTIONAL COMMITMENT

The Dean of the School of Medicine, Steve Nelson, MD, has agreed to continue providing partial financial support for the CIMC facilities now and after the COBRE Phase III ends. Moreover, Dr. Nelson will facilitate the integration of the CIMC and other COBRE cores into the research structure of LSU-HSC. After completion of this Phase II of the COBRE, we expect that the MHAM will be funded by the user fees (approximately 30%), LSU Cancer Center funds (40%) and School of Medicine funds (30%). An example of the strong institutional support is the addition of our new state-of-the-art confocal microscope, which was purchased with LSU and Cancer Center funds.

FACILITIES

Laboratory: The CIMC is located on the 7th floor of the new state of the art Cancer Center building, LSU Health Sciences Center campus, at 1700 Tulane Avenue, in New Orleans, LA. 70112. It consists of two laboratories, one dedicated to the Molecular Histopathology section of the core, and the other one for the Analytical Microscopy section, and totaling 400 square feet. Dr. Del Valle’s laboratory and office, as well as Drs. Parker Struckhoff and Trillo-Tinoco’s cubicles are located on the same floor. Common lab equipment includes two 4°C refrigerators (VWR GDM47), one -20°C freezer (Frigidaire FKCH17F7WC), two -80°C freezers (Thermo Scientific Forma 989), one refrigerated microfuge, two thermocyclers (BioRad), two waterbaths, a sonicator, a UV/Vis spectrophotometer, a BioRad micro-plate reader, a pH meter, a scale and balance, a platform shaker, three centrifuges, and one conventional incubator for bacterial cultures.

Computers: The CIMC laboratory has one Apple Pro computer, three Optiplex 620 Dell computers and two Optiplex 740 Dell computers, as well as 2 laser color printers and one scanner.  The laboratory is fully connected to the Internet and the LSUHSC mainframe computers.

SPECIFIC AIMS

The Molecular Histopathology and Analytical Microscopy Core (MHAM) is a new scientific core, with a mission to assist investigators of this COBRE, and investigators in the region, in the detection, imaging, and analysis  of gene and protein dynamics in any type of cellular models and cancer tissue (animal or human), while maintaining high quality, consistent reproducibility, and technical expertise for microscopy and histopathological studies. Histochemical and immunohistochemical methodologies will be performed to provide information on protein expression, localization and interactions between viral and cellular proteins. Molecular histopathology, including in situ hybridization will be performed to provide investigators with information regarding cellular and viral DNA and/or RNA. Another crucial objective of the core is to provide support to the investigators of individual pilot projects in successfully obtaining extramural funding. We will also provide access to clinical samples to promote the translational aspect of the research program that is crucial to investigate how molecular and cellular pathways identified in vitro or in experimental animals correlate with human disease.

For this purpose, Molecular Histopathology is equipped with a complete tissue-processing center while Analytical Microscopy is equipped with upright bright field and epifluorescence microscopes with digital cameras, an inverted fluorescence microscope for Comet assay, a fluorescence microscope with  deconvolution software, and a state of the art confocal microscope. The core also has a Laser Micro- Dissecting microscope for single cell dissection. Various histological techniques are offered and pathology expertise for the interpretation of results and generation of data and figures is provided.

To support these goals the MHAM Core will carry out the following Specific Aims:

Specific Aim 1. Provide high quality molecular histopathology and microscopy imaging on murine and human tissues to support research activities, and to generate high-quality images and figures for grant proposals and presentations for researchers in this COBRE and in the region.

Specific Aim 2. Develop advanced molecular techniques in tissue imaging as they become available including in situ Proximity Ligation technology (DuoLink) in combination with in situ hybridization. In addition provide high quality immunohistochemical experiments with antibodies for cellular markers, viral proteins and specific pathway proteins, double and multiple immunofluorescence labeling, in situ hybridization with DNA and RNA probes, and Fluorescence in situ Hybridization (FISH).

Specific Aim 3. Provide pathological evaluation and analysis for human patient samples, as well as support in the interpretation of the results generated by specialized imaging analyses, including deconvolution, confocal and time-lapse imaging.

Specific Aim 4. Collect and archive clinical cases (autopsy and biopsy samples) from patients with different tumors and matching normal tissue controls. This task will be performed as a collaborative effort with HIV/AIDS Biorepository at LSU Cancer Center and the Tumor Biorepository of the LCRC.

Specific Aim 5. Mentor young investigators and students, and advise senior investigators, on the application  of microscopic techniques, histological analyses and immunohistochemistry to enhance their research projects and to provide avenues towards developing translational research projects.

The MHAM Core works in concert with the Cellular Immunology and Immune Metabolism Core and the Translational Genomics Core, ensuring the development of parallel complementary techniques to image and detect the DNA, RNA or proteins of interest such that the investigators will have comprehensive data from in vitro cultures, animal experiments, and human tissues available to support their work.

PLANNED APPROACHES

Dr. Luis Del Valle will direct all aspects of the MHAM Core including providing scientific direction, organizing and prioritizing activities of the core, assigning duties, and ensuring a high quality in the output of the results. Dr. Del Valle will also sign off on any results being provided to the researchers, in particular any work requiring pathological analysis of human samples. The three members of the core (Drs. Del Valle, Parker Struckhoff, and Trillo-Tinoco) inspect all microscopes on weekly basis in order to determine maintenance needs and other potential issues. For any potential problems, the service contracts from the equipment providers cover the annual inspection of each microscope, and in certain cases the costs of repairs and upgrades.

Dr. Del Valle will resolve personnel issues and address potential conflicts with users of the Core. Finally, Dr. Del Valle will be responsible for reporting on the activities of the core to the PI and to the Steering Committee during the quarterly meetings, and he will be responsible for preparing and presenting an annual review on the MHAM Core for the External Advisory Board and for the annual progress report to the NIH.