Hostname: page-component-cd9895bd7-dzt6s Total loading time: 0 Render date: 2024-12-27T10:09:28.752Z Has data issue: false hasContentIssue false
  • English
  • Français

Resolution in Light Microscopy

Published online by Cambridge University Press:  02 July 2020

H. Tandler ,
L. Schreiber ,
U. Simon  and
H. E. Keller
H. Tandler
Affiliation:
Carl Zeiss GmbH Microscope DivisionD-07740Jena, Germany
L. Schreiber
Affiliation:
Carl Zeiss GmbH Microscope DivisionD-07740Jena, Germany
U. Simon
Affiliation:
Carl Zeiss GmbH Microscope DivisionD-07740Jena, Germany
H. E. Keller
Affiliation:
Carl Zeiss GmbH Microscope DivisionD-07740Jena, Germany
Get access

Extract

The resolution in conventional light microscopy was defined by Abbe as: Wavelength (λ) and Numerical Aperture (NA) have a direct influence while the factor (c) is dependent upon the geometry of the illumination and observation beam paths. Some typical examples are:

High illumination apertures in darkfield or oblique illumination or specific ratios between illumination and observation aperture can substantially improve resolution. Various shapes of the illumination aperture, annular, point, centered or off axis at different azimuths directly influence numerical aperture and coherence and with this resolution, contrast and depth.

In mirror objectives low apertures become vignetted, the resolution is enhanced at the expense of contrast. In studies of Hopkins and Barham the factor “c” can range from 0.5 to 0.8. For the practically attainable condition, where illumination and observation aperture are identical, “c” becomes 0.61 and we have partial coherence.

Exceeding the observation aperture on the illumination side reduces “c” further but diffraction significantly reduces contrast.

Type
The Limits of Image Resolution: Seeing is Believing
Information
Microscopy and Microanalysis , Volume 3 , Issue S2: Proceedings: Microscopy & Microanalysis '97, Microscopy Society of America 55th Annual Meeting, Microbeam Analysis Society 31st Annual Meeting, Histochemical Society 48th Annual Meeting, Cleveland, Ohio, August 10-14, 1997 , August 1997 , pp. 1175 - 1176
Copyright
Copyright © Microscopy Society of America 1997

Access options

Get access to the full version of this content by using one of the access options below. (Log in options will check for institutional or personal access. Content may require purchase if you do not have access.)