Practical limits of resolution in confocal and non‐linear microscopy

Abstract
Calculated and measured resolution figures are presented for confocal microscopes with different pinhole sizes and for nonlinear (2‐photon and second harmonic) microscopes. A modest degree of super‐resolution is predicted for a confocal microscope but in practice this is not achievable and confocal fluorescence gives little resolution improvement over widefield. However, practical non‐linear microscopes do approach their theoretical resolution and therefore show no resolution disadvantage relative to confocal microscopes in spite of the longer excitation wavelength. Microsc. Res. Tech. 63:18–22, 2004.

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