Real time imaging of the detection volume of a confocal microscope

Deconvolution, pixel reassignment or adaptive optics-based strategies utilize information about the detection profile in improving the resolution of optical microscopy. Here, we show a novel method which allows us to obtain the single-photon detection volume of a laser scanning confocal microscope at any desired location of the object. It can create a stationary, virtual ‘guide star’ at the chosen location while the excitation beam is scanning the sample, by using an optical fiber placed in the non-descanned path of the microscope. Our experimental results are verified by diffraction theory-based calculations. The major advantages of our method are that it is alignment free, affordable, sensitive and applicable to many different modes of confocal imaging.

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