Micropatterning on EM grids: A strategy for improving cell cryo-ET workflow
Tuesday, October 15, 2019
9AM PDT | 12pm EDT | 5pm BST | 6pm CEST
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Micropatterning of extracellular matrix proteins is an established in vitro cell culture technique for controlling the structure of the cell microenvironment. In recent mechanobiology studies, micropatterning has been used in conjunction with traction force microscopy to demonstrate correlation between cell morphology and the regulation of force transmission. A current challenge, however, is understanding how cells sustain increased strain energy states and localized stresses at the supramolecular level. Cryogenic electron tomography is the highest resolution tool available for structural analysis of macromolecular organization inside cells. In whole cell cryogenic electron tomography, cells are cultured on specialized substrates, called electron microscopy grids, before being vitrified and loaded into an electron microscope.
The speaker presents a method in which the PRIMO maskless protein photopatterning system is used to micropattern extracellular matrix on electron microscopy grids. These micropatterned cell culture substrates can confine cells to specific shapes and locations on electron microscopy grids. By enabling cryo electron tomography of cells under morphological modulation, this technology may open up new possibilities for correlating changes in nanometer-scale organization at cell-cell and cell-ECM contacts to strain energy states and traction stress distribution in the cell.
- How to structure in vitro cell microenvironments on electron microscopy grids.
- How to reproducibly position cells cultured on EM grids.
- The advantages of maskless photopatterning with PRIMO for micropatterning EM grids