Pathology using immunohistochemistry (IHC) remains a critical tool in modern clinical medicine. However, the capabilities of routine IHC methods have not kept pace with advances in biomedicine, which require evaluating networks of, e.g., tens of large molecules (typically protein targets) in a tissue context by multiplexing. To overcome this limit, highly multiplexed IHC imaging systems based on mass spectrometry and repeated Immunofluorescence (IF) imaging have been developed. These systems claim theoretical multiplexing of up to 100 targets. However, they are too slow, expensive, and complex, for routine use, and are currently used in a small number of research laboratories. We are working to radically change the capabilities of IF imaging to allow routine detection of tens of molecular targets rapidly and at moderate cost. Our approach builds on our previous work on high-speed hyperspectral imaging coupled with amplified molecular target labeling methods. We will advance our technology and demonstrate its capabilities with a specific research application involving imaging of brain sections from a mouse model of a neuropathology relevant for Alzheimer's disease (AD). We will make improvements to achieve simpler and faster operation of our technology. We will then image up to 12 targets and demonstrate the ability to extend our approach to tens of targets on a single tissue slide. We expect our instrumentation and protocols will be sufficiently simple, low cost, and rapid to allow routine use in most pathology and research laboratories. !
Public Health Relevance Statement: PROJECT NARRATIVE This project will extend the high multiplexing capabilities, currently available in complex mass spec and repeated immunofluorescence (IF) imaging methods to a simple, low cost and rapid IF approach, in turn enabling widespread and routine use of multiplexed IF in pathology. In the near term, our approach will provide a powerful imaging tool to better inform research on a wide range of diseases. In the longer term, the approach will be extended to clinical pathology, where it could support better informed diagnosis and therapy selection for neurodegenerative, neoplastic, and other diseases. !
Project Terms: AD dementia; Alzheimer Type Dementia; Alzheimer disease dementia; Alzheimer sclerosis; Alzheimer syndrome; Alzheimer's; Alzheimers Dementia; Primary Senile Degenerative Dementia; primary degenerative dementia; senile dementia of the Alzheimer type; Alzheimer's Disease; Antibodies; Brain; Brain Nervous System; Encephalon; Cell physiology; Cell Function; Cell Process; Cellular Function; Cellular Physiology; Cellular Process; Subcellular Process; Clinical Medicine; Clinical Medical Sciences; Complement; Complement Proteins; Data Collection; Diagnosis; Disease; Disorder; Goals; Immunohistochemistry; Immunohistochemistry Cell/Tissue; Immunohistochemistry Staining Method; instrumentation; Laboratories; Laboratory Research; Lighting; Illumination; Literature; Methods; Transgenic Mice; Mitochondria; mitochondrial; Modernization; Mus; Mice; Mice Mammals; Murine; Nerve Degeneration; Neuron Degeneration; neural degeneration; neurodegeneration; neurodegenerative; neurological degeneration; neuronal degeneration; Neuroanatomy; Neuranatomies; Neuranatomy; Neuroanatomies; Pathology; Clinical Pathology; Proteins; Research; Ribonucleoproteins; RNA Splicing; Splicing; Role; social role; Mass Spectrum Analysis; Mass Photometry/Spectrum Analysis; Mass Spectrometry; Mass Spectroscopy; Mass Spectrum; Mass Spectrum Analyses; Technology; Testing; Time; Tissues; Body Tissues; Translations; translation; Work; Immunofluorescence Immunologic; Immunofluorescence; Paraffin Embedding; selection of treatment; therapy selection; treatment selection; Selection for Treatments; Label; Microscope; sensor; improved; Image Analyses; image evaluation; image interpretation; Image Analysis; brain visualization; Brain imaging; Clinical; Phase; TAR DNA-binding protein 43; TDP-43; TDP43; protein TDP43; protein TDP-43; tool; instrument; Hour; Complex; Scanning; Protocols documentation; Protocol; Slide; System; brain tissue; meter; Nuclear; Neurodegenerative Disorders; Degenerative Neurologic Diseases; Degenerative Neurologic Disorders; Nervous System Degenerative Diseases; Neural Degenerative Diseases; Neural degenerative Disorders; Neurodegenerative Diseases; Neurologic Degenerative Conditions; degenerative diseases of motor and sensory neurons; degenerative neurological diseases; neurodegenerative illness; fluorophore; RNA Transport; Ribonucleic Acid Transport; Speed; novel; Reporting; Pathology Report; Modeling; neuropathology; neuropathologic; neuropathological; image-based method; imaging method; imaging modality; Detection; Imaging Device; Imaging Instrument; Imaging Tool; Molecular Target; Resolution; resolutions; in vivo; Collection; Experimental Pathology; Development; developmental; Image; imaging; neoplastic; age dependent; age related; cost; optic imaging; optical imaging; Consumption; Coupled; murine model; mouse model; fluorescent imaging; fluorescence imaging; bio-markers; biologic marker; biomarker; Biological Markers; operations; operation; imaging platform; imaging system; Multiplexed Ion Beam Imaging; multiplexed imaging; imaging mass spectrometry; mass spectrometric imaging
