Background

In 4Pi STED scanning fluorescence light microscopy, it is known to use devices for forming three-dimensional light intensity distribution comprising a spatially limited area of minimum light intensity that is enclosed by areas of higher light intensity of the fluorescence inhibiting light (a).

This technology provides an apparatus for 4Pi STED which allows the generation of a three-dimensional light intensity distribution comprising a very steep light intensity gradient between the areas of minimum and maximum light intensity.

Technology Overview

The apparatus comprises two objectives which are facing each other on a common optical axis, which focus light coming out of opposite directions into a common focal area, and which each have a pupil, and at least one light source. In case of three or more pairs of coherent light beams, the area of minimum light intensity is reduced to a small volume within the focal area (Fig. 1c).

Stage of Development

According to numerical calculations in a realistic scenario even DVD diodes as STED lasers will result in highest resolutions in the 20nm range and below in all three dimensions.

Benefits

• Technology can be used for 4Pi STED, RESOLFT, and 4Pi GSD
• Precise three-dimensional light intensity distribution
• Steep light intensity gradient

Applications

The presented technology allows for the generation of an even steeper light intensity gradient between the area of minimum light intensity to the enclosing areas of higher light intensity than in isoSTED and can be used for 4Pi STED, RESOLFT or another variant of 4Pi GSD scanning fluorescence light microscopy.