The CLEM Workflow
The further we dive into multimodal imaging, the least it becomes possible to define correlative microscopies.
To cut a long story short, we aim to observe one exact same sample sequentially using various imaging technologies, to ultimately blend them into one single virtual dataset.
To achieve that, the sample must be prepared from the very first step with all the various transformations it will undergo in mind.
We focus our attention on Fluorescence Live Cell Imaging, cryo-immobilization by high pressure freezing, electron microscopy imaging and ultimately, image registration. This is a narrow path, but likely the best to cover the broadest range of multimodal cell and tissue imaging. Want to know more? Let's take an example
In this video, we showcase our live CLEM-HPF workflow by capturing a complete experiment, ranging from live cell imaging and cell viability control using a standalone microscope to the integration with our HPM-Live µ for live cell imaging of a sample prior to HPF.
Live Cell Imaging
Life science is about observing changes in a dynamic environment upon stimulation to understand the underlying mechanisms. Live cell imaging is therefore one major technique used to observe these changes as they happen. To make sure we capture the event of interest as we go to the electron microscope (a tool to observe ultrastructure of fixed material), we associate a light microscope to our High Pressure Freezer to achieve a temporal resolution of 1.26second between the last living image and the cryo-immobilization of the sample.
High Pressure Freezing
Multimodal imaging often requires observing the sample through physically distinct microscopes, or after applying a physical transformation of the sample between two imaging steps.
Image registration is therefore an essential step where accurate superimposition of the various images at distinct magnification will allow targeting the structure of interest and identify precisely the phenotype of interest.