Our overarching goal is to study how cells of the immune system and glial cells within the central nervous system contribute to the development and progression of neurological disorders such as multiple sclerosis. Immunofluorescent microscopy is a standard and widely used technique across biological disciplines and is essential to neuroscience and immunology research. One of the main issues that plague researchers is that immunofluorescent microscopy is typically limited in the number of probes that can be imaged at a single time due to the limitations of conventional microscopes.
In this protocol, we describe a method to expand the number of probes that many microscopes can image at a single time, and provide an analysis pipeline to process information-dense images. The advantage of this protocol is that it can be adapted to many widely available microscopes and is easy to execute, which would make multiplex histology analysis available to a greater number of labs.
