Putting Dynamic Live Cell Data into the Ultrastructural Context

Now there is no need to guess about the underlying mechanisms when analyzing a wide range of biological events, like intracellular trafficking, nuclear envelope formation, or mitotic spindle assembly. The workflow Coral Life is a proven solution, combining live-cell correlative light and electron microscopy (CLEM) with time-specific fixation by high-pressure freezing, which enables the investigation of dynamic events with nanometer resolution. A massive surplus of context data can be obtained by the combination of dynamic data, specifically targeted entities by fluorescent live-cell imaging, and precisely timed EM ultrastructural analysis.

Many electron microscope workflows start with the fixation of cells, followed by sample preparation and EM imaging. However, cells displaying interesting behavior tend to be rare. Making things even more complicated is the fact that the cellular behavior or processes of interest might be very transient. Finding the “right cell” is often very time-consuming and tedious work which seems like “looking for a needle in a haystack”.

The Live-cell CLEM workflow allows you to capture dynamic information of a relevant biological process when it happens and enables you to put these observations into their ultrastructural context. With Coral Life, you can rest assured that you obtain the results you are looking for. Find the right cell in the right state at the right time with the light microscope, observe it, and retrieve it rapidly and reliably once the sample is loaded in the electron microscope. Thanks to the dedicated SampLink chamber, you enjoy peace of mind while looking for the cell of interest with the light microscope and quickly “snap-freeze” it precisely at the correct moment.

A transfer time of just 5 seconds opens a window to a wide range of dynamic processes which occur in living cells. With some experience, you can even refine the exact point in time of the processes which you want to capture in the prepared specimens. Whether you are interested in vesicle transport and sorting, filopodial dynamics, endocytosis, cell migration, cytokinesis, or a plethora of other biological processes, Coral Life gives you complementary dynamic and ultrastructural information.

The generous time window before the transfer ensures that you to have enough room for sample identification and imaging. Without the pressure to make a fast decision, typical of other live-cell CLEM systems where often there are only a few minutes to choose a region, you can make the selection more calmly, the success rate increases, and the results are of better quality and more reproducible.