Evoking The Full Potential of Confocal Microscopy
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Confocal laser scanning microscopes (CLMs) are renowned for their sectioning capability. This feature is enabled by the use of pinhole, which rejects out-of-focus light. Less appreciated, on the other hand, is the gain in lateral resolution by this type of microscopes for one obvious reason.
As the pinhole is closed to improve the resolution, less light can reach the detector leading to images with poor signal-to-noise rations (SNR). Therefore CLSMs are operated with a pinhole diameter of around 1 Airy unit (AU), sacrificing resolution for SNR. But with a clever detector design one can solve this conundrum.
By using a sub-Airy detector element array instead of a single point detector, it is possible to collect light more efficiently boosting the strenghts of confocal. The extra photon budget can be diverted towards increasing the sensitivity, the scanning speed or the resolution of the microscope.
In this article we will focus our discussion on how resolution is improved.