Mental and neurological disorders account for a large part of the total global disease burden, yet there is a severe lack of effective treatments for reducing the associated disability and mortality. Brain dysfunctions are caused by a large variety of factors, such as pathological network connectivity, altered cellular and physiological properties, and neurotransmitter imbalances that act together or alone to result in profound behavioral impacts. Thus, there is an urgent need for integrative tools that allow an unbiased whole-brain understanding of the underlying pathophysiology of complex brain disorders. Recent advances in tissue clearing, labeling, and high-resolution light sheet microscopy, are enabling mapping and comparative analysis of large intact brain samples in normal and diseased states. However, multiple challenges remain, specifically in achieving uniform labeling of specific molecular targets in large tissues, scalable microscopy platforms for high-resolution whole-brain imaging, and multi-scale high-accuracy comparative data analysis tools. Here, I present my work in the development of a set of novel methods to address some of these challenges.
The first aim focuses on developing a rapid and uniform deep tissue molecular labeling method by utilizing modified DNA aptamers to significantly reduce the staining times (e.g., less than 4 days for an intact mouse brain, as opposed to several weeks). The second aim introduces a cost-effective (~20x cheaper) and scalable light sheet fluorescence microscopy (LSFM) implementation, so-called projected Light sheet microscopy (pLSM), for rapid high-resolution imaging of large biological samples. The third aim is focused on developing a suite of large data analysis methods (suiteWB) for high-resolution whole-brain comparative phenotyping – both at the level of neuron densities and their brain-wide projection patterns. Through this pipeline, we systematically investigated the brain-wide dopaminergic modulatory pathway alterations resulting from chronic ketamine exposure.
Altogether, these sets of highly integrative labeling, imaging, and analysis tools will facilitate a comprehensive understanding of the pathophysiology of complex brain disorders and the discovery of novel therapeutic targets.
| Identifer | oai:union.ndltd.org:columbia.edu/oai:academiccommons.columbia.edu:10.7916/mgmg-ee39 |
| Date | January 2024 |
| Creators | Chen, Yannan |
| Source Sets | Columbia University |
| Language | English |
| Detected Language | English |
| Type | Theses |
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