Flow Cytometry
We use flow cytometry, also known as FACS, to characterize the expression of inflammation markers by neuronal and immune cells.
Flow cytometry is a laser-based technology employed in cell counting, cell sorting, and biomarker detection by simultaneously analyzing multiple physical and chemical characteristics in thousands of particles per second.
Calcium Imaging
See: https://www.jove.com/science-education/5203/calcium-imaging-in-neurons
Calcium imaging is used to characterize neuronal responses to different stimuli, including the signals produced by the cells of the immune systems. Fluorescent changes in responses to stimuli are recorded from neurons that are genetically encoded or loaded with a calcium indicator.
Disease modeling with mice genetically-engineered
We used the cre/lox toolbox to create mice whose sub-populations of neurons are genetically engineered to be ablated (lsl-DTA), silenced (lsl-TeTx), fluorescent (lsl-GFP), activity-dependent reporter (lsl-GcamP6), or light sensitive (lsl-ChR2).
Primary neuron and immune cell culture
Typically we model diseases in mice, and harvest their neuronal and immune cells to assess their individual properties (sensitivities, responsiveness, activation level) and how they interact with one another.
Imaging. 2P, confocal, fluorescent and IHC.
Protein expression is detected on different tissue using specific antibodies and imaged using confocal or fluorescent microscope.
Expression profiling.
Single cell qPCR. In situ hybridization and RNA sequencing.
Genes transcripts are assessed at the single cell level using qPCR, in situ hybridization and RNAseq.
Optogenetic
Adapted from: https://commons.wikimedia.org/wiki/File:Optogenetic_stimulation_consists_of_several_steps.png
Optogenetic used mice genetically-engineered to express light sensitive protein and allow thecontrolled activation of neurons susbets with an unparalled spatio-temporal resolution.