Practical limits of resolution in confocal and non-linear microscopy.
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.
Cox
Guy
G
Instituto Gulbenkian de Ciencia, Oeiras, Portugal. guy@emu.usyd.edu.au
Sheppard
Colin J R
CJ
D000818:Animals; D003094:Collagen / Q000648:ultrastructure; D020405:Dermis; D007614:Macropodidae; D051379:Mice; D018613:Microscopy, Confocal / Q000379:methods*; D036641:Microscopy, Fluorescence, Multiphoton / Q000379:methods*; D017711:Nonlinear Dynamics; D012680:Sensitivity and Specificity; D013623:Tail; D013710:Tendons / Q000648:ultrastructure
D003094:Collagen