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Lloyd Cantley (PI)
Kidney function, both under normal conditions and in the setting of disease, is predominantly controlled by the amount, localization and activation state of proteins in specific cells at specific sites and under specific spatial influences that in turn regulate cell differentiation, division, metabolism, morphology, membrane polarization, secretion and transport. Imaging mass cytometry (IMC) takes advantage of laser ionization and time of flight mass spectrometry to simultaneously identify up to 42 metal ion-conjugated antibodies from tissue sections at high spatial resolution with essentially no background signal. In the current proposal, human kidney samples will be used to develop protocols for high-fidelity staining with individual antibodies to identify cell types and protein activation states of interest in the kidney (SA 1) followed by staining of normal and diseased human kidney sections with pooled, metal ion-conjugated antibodies for multiplex IMC image reconstruction to simultaneously define the activation/expression states of multiple cells in the human kidney (SA 2).