3D imaging of living cells can be challenging for a number of reasons, such as low signal-to-noise ratio, reduced cell viability and photobleaching of fluorophores.
The crucial role of choosing the right immersion oil is often overlooked.
Conventional immersion oil matches the refractive index of glass – but not of the sample. As a result, conventional oil works well when imaging near the surface, but when moving deeper into the sample it can introduce spherical aberration, which creates spatial distortions leading to elongated 3D reconstructions.
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