Improved resolution in "practical" light microscopy by means of a glass fiber 2 pi tilting device

Joachim Bradl; Bernd Rinke; Bernhard Schneider; Michael Hausmann; Christoph G. Cremer

doi:10.1117/12.229984

15 January 1996 Improved resolution in practical light microscopy by means of a glass-fiber 2 π-tilting device

Joachim Bradl, Bernd Rinke, Bernhard Schneider, Michael Hausmann, Christoph G. Cremer

Proceedings Volume 2628, Optical and Imaging Techniques for Biomonitoring; (1996) https://doi.org/10.1117/12.229984
Event: BiOS Europe '95, 1995, Barcelona, Spain

Abstract

The spatial resolution of a conventional light microscope or a confocal laser scanning microscope can be determined by calculating the point spread function for the objective used. Normally, ideal conditions are assumed for these calculations. Such conditions, however, are often not fulfilled in biological applications especially in those cases where biochemical requirements (e.g. buffer conditions) influence the specimen preparation on the microscope slide (i.e. 'practical' light microscopy). It has been shown that the problem of a reduced z- resolution in 3D-microscopy (optical sectioning) can be overcome by a capillary in a 2(pi) - tilting device that allows object rotation into an optimal perspective. The application of the glass capillary instead of a standard slide has an additional influence on the imaging properties of the microscope. Therefore, another 2(pi) -tilting device was developed, using a glass fiber for object fixation and rotation. Such a fiber could be covered by standard cover glasses. To estimate the resolution of this setup, point spread functions were measured under different conditions using fluorescent microspheres of subwavelength dimensions. Results obtained from standard slide setups were compared to the glass fiber setup. These results showed that in practice rotation leads to an overall 3D-resolution improvement.

Citation Download Citation

Joachim Bradl, Bernd Rinke, Bernhard Schneider, Michael Hausmann, and Christoph G. Cremer "Improved resolution in practical light microscopy by means of a glass-fiber 2 π-tilting device", Proc. SPIE 2628, Optical and Imaging Techniques for Biomonitoring, (15 January 1996); https://doi.org/10.1117/12.229984

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