Obtain better cell samples for your cryo-electron tomography experiments !
Cryo-EM is rapidly expanding for imaging intracellular molecules conformations. However, imaging cell samples with cryo-ET can be a challenge. Indeed, before even considering going to the microscope, the cell sample must meet some basic requirements: matching specific locations on the EM grid, containing the target component, being thin enough for transmission electron microscopy.
By controlling the cell adhesion, spreading and shape, PRIMO maskless and contactless micropatterning overcomes the issues faced at the very first step of the workflow of cell sample preparation for cryo-ET.
It provides unique advantages for your whole-cell TEM and cryo-ET experiments:
- Precise cell positioning: automatic alignment within the mesh of EM grids!
- Optimized cell spreading: reduced thickness for more efficient CLEM and cryo-ET workflows
- Standardized cell models: reproducible micropatterning for mechanobiology studies
- Undamaged surface: contactless micropatterning preserving the EM grid surface integrity
Cell positioning within EM grid mesh
Leonardo software detects the mesh of EM grids and automatically aligns your patterns within it, for PRIMO to perform aligned micropatterning. The PRIMO system thus ensures cells will be well positioned within the mesh of your EM grids !
And thus make sure you get cells optimally positioned for FIB milling and cryo-ET !
Optimized cell spreading on EM grids thanks to micropatterning
By controlling the available adhesive surface and cell adhesion on EM grids, PRIMO micropatterning can modulate the cell spreading. And the cell spreading in turn can be a key asset for improved throughput for whole-cell cryo-electron tomography. Indeed, the spreading facilitates the identification (CLEM), preparation (FIB milling, sometimes not even needed if the sample is thin enough) and imaging (cryo-ET) of the intra-cellular targeted structures.
Micropatterning for standardized cell models
Micropatterning is known as an effective way to control cell adhesion in vitro. This technique is therefore largely used for standardizing cell shape as well as cell internal organization (directly linked to external adhesive cues).
With its unique capacity to automatically align micropatterns within the mesh of EM grids, PRIMO micropatterning appears as the perfect tool for conducting homogeneous and standardized cryo-EM experiments.
To go further, using PRIMO micropatterning for cryo-ET experiments on cells could help precisely target specific internal elements when correlative imaging approaches prove to be too challenging, as emphasised by M.
EM grid undamaged surface
As a maskless and contactless photopatterning system, PRIMO can project your micropatterns in UV light on the surface of EM grids without compromising their integrity!
Ressources: Using the PRIMO micropatterning system for cryo-ET
Find below several support tools to help you further investigate the use of the PRIMO micropatterning system to facilitate cryo-ET experiments.
