ShEEP Request for a Zeiss Axio Imager M2 with Stereo Investigator System

Stroke, traumatic brain injury (TBI), cardiac arrest (CA), posttraumatic epilepsy (PTE), and neuropathic pain are leading causes of death and long-term disability among Veterans. This ShEEP proposal is to request purchasing the Zeiss AxioImager M2 ApoTome with Stereo Investigator System (# 27552 - MBF Bioscience) for quantitative and microscopic imaging analysis of mounted slide tissue using

Design Based Unbiased Stereology protocols. This is a complete system with the updated ApoTome which can achieve a comparable or higher axial resolution than that of high-end confocal laser scanning microscope (cLSM). With the advent of high-resolution 2- and 3-dimensional microscopic digital imaging

systems, the design-based unbiased quantitative analysis has become the biggest obstacle for comparing the focal and cell-specific morphological and biochemical/molecular changes in animal disease models. This is because of uneven distribution of histological staining and antibody-based immunolabeling.

To tackle these quantitative challenges, MBF Bioscience has developed the best Zeiss AxioImager M2 ApoTome with Unbiased Stereology System. The techniques supported by this system are the most efficient and accurate in the industry. The Zeiss AxioImager M2 ApoTome with Stereo Investigator System has no special facility requirements; and will be utilized mainly by five users but will be available to all VA

researchers. This system can produce quantitative microscopic imaging data for the statistical analysis rigor and clear reporting in scientific publications. Therefore, this system will significantly improve our ability for quantifying a wide variety of numerical imaging parameters such as: size, shape and protein

compositions by robust, efficient, quantitative scoring at the subcellular and macromolecular levels on tissue sections using stroke, CA, TBI, PTE and neuropathic pain animal models. Four VA-funded projects are included in this application. The Baltimore VA currently does not have a quantitative imaging system.

There are already close interactions and collaborations among the users, and this instrument will further foster such interactions and collaborations. Currently, the Baltimore VA Research Core is directed by Joungil Choi, PhD, who has wide expertise in biochemistry, histology, immunoassay development, and

molecular biology. She will work closely with Dr. Bingren Hu (PDF/PI) to develop standard operating procedures and ensure that the personnel are well trained, and the instrument is maintained properly. Overall, this Zeiss AxioImager M2 ApoTome with Unbiased Stereology Platform is highly suitable for a

group of researchers who need to perform unbiased quantitative microscopic imaging analysis of cells and tissue sections as tools for biomedical research. All VA-funded participants for this ShEEP application have extensive needs of using the unbiased quantitative microscopic imaging analysis for: brain injury volume,

number of injured cells, morphological and protein composition changes in subcellular structures, inflammatory biomarkers and signaling molecules on tissue sections of different animal models. Without this equipment, these VA-funded studies will have significant limitations in performing unbiased quantitative

microscopic imaging analysis. Therefore, the Zeiss AxioImager M2 ApoTome with Unbiased Stereology Platform will provide a state-of-the-art tool for VA-funded projects to study disease mechanisms, diagnosis, and potential treatments to improve Veterans’ health and well-being.