Neurological disorders such as Alzheimer's and Huntington's disease, spinal cord injury, traumatic brain injury, multiple sclerosis, radiation injury, and brain developmental disorders are characterized by profound neuronal and glial injury, leading to significant impairments in cognitive and/or motor functions. Although the etiology of these disorders vary, the signature and defining pathological features for many of these conditions is the loss of neurons and synapses, loss of neuronal connectivity, tissue volume loss and expansion of the ventricular spaces, dysregulated neurogenesis, and rampant neuroinflammation, gliosis, and scarring. The investigators listed on this proposal are all members of the Institute for Memory Impairments and Neurological disorders at UC Irvine (UCI MIND), and are seeking funds to acquire a multichannel fluorescent structured illumination stereological imaging system to quantify tissue changes with unbiased methods. Neuropathological studies are severely limited by attempts to study three-dimensional interactions by observations made in two dimensions. Stereology, however, is a science that overcomes these limitations by furnishing three- dimensional interpretations of planar sections of biological tissues, as it is based on merging of the principles of geometry and statistics. Consequently, stereological studies of the nervous system provide a means to obtain more precise quantitative measurements of cell number and neural processes and tissue volume. In addition, for studies on neurodegenerative disorders, it is possible to determine the quantitative relationship between the accumulation of brain lesions such as amyloid plaques and cell number, for example. Standard brightfield stereology allows one to accurately quantify one immunolabel at a time in tisue sections. Double and/or triple labeling immunocytochemistry typically require fluorescent labeling and more specialized imaging techniques. An Apotome structured illumination system allows confocal-like 3D images to be acquired for both real-time and off-line analysis of multiple labels. Such a system would allow investigators to quantify two or three parameters simultaneously (e.g. total number of GFP-expressing transplanted cells, and a second or third fate marker to obtain total engraftment numbers and percentage of neural, astroglia and oligodendroglial fate in the same tissue sections).
