Abstract
The imaging performances of multiphoton excitation and confocal laser scanning microscopy are herby considered: in typical experimental imaging conditions, a small finite amount of photon reaches the detector giving shot-noise fluctuations which affects the signal acquired. A significant detriment in the high frequencies transmission capability is obtained. In order to partially recover the high frequencies information lost, the insertion of a pupil plane filter in the microscope illumination light pathway on the objective lens is proposed. We demonstrate high-frequency and resolution enhancement in the case of linear and non linear fluorescence microscope approach under shot-noise condition.
Highlights
Shot-noise represents a pernicious limit in scanning fluorescence microscopy, significantly affecting and deteriorating the imaging capabilities[1,2]
This study reveals a remarkable shot-noise influence in the high-frequency range with a relevant detriment of the high-frequencies transmission capabilities and a substantial reduction of the optical transfer bandwidth
In order to partially retrieve the high frequencies information loss, we propose to insert an annular filter on the microscope objective lens in the illumination light pathway[3,4]
Summary
Shot-noise represents a pernicious limit in scanning fluorescence microscopy, significantly affecting and deteriorating the imaging capabilities[1,2]. Adopting computational simulation methods, a detailed analysis of its influence in typical two-photon excitation (2PE) and confocal laser scanning microscope (CLSM) imaging conditions is here presented, both in the spatial and in the frequency domain.
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