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An Objective Lens with an Incorporated Beam Splitter for Multiphoton Microscopy

Others

*Abstract

Compound lens incorporating dichroic elements that effectively decouples excitation and emission paths within the objective lens, allowing increased field of view, more efficient light collection, and a longer working distance.

Commercial innovation in objective lenses, especially in the field of laser scanning microscopy, has focused on increasing numerical aperture. As resolution of objective lenses increases, field of view generally decreases, for multiphoton imaging, FOVs less than one millimeter are not uncommon.

Increasingly however, confocal and 2 photon microscopes are being used to image tissue, with an eye toward clinical applications. In these applications one is willing to sacrifice resolution for an increase in FOV, more efficient light collection, and a longer working distance.

Warren Zipfel's lab at Cornell has designed new lenses for laser scanning microscopy of tissue. The lenses have been optimized for:

Larger field of view

4 mm

Longer working distance

2 mm

Sufficient NA for optical sectioning and single cell resolution

~1

Maximize light collection

Collects ~10x more fluorescence than the standard 4X macro lens.

Chromatic correction over bandwidth of typical fs pulse

Corrected over 10- 15 nm

Axial FWHM of the focal volume

10 microns

Inexpensive design to fabricate

- only two glass types

- no aspheric surfaces

A key innovation, is the incorporation of dichroic element into the lens itself, which relaxes the need to collimate collected light. This allows for an increase in the size of the front aperture of the objective as well as the application of a reflective coating to the interior of the objective casing, both of which boost the collected solid angle far in excess of what one would predict based on NA alone. The design effectively decouples the excitation and emission paths within the objective lens.

 

Keywords: Physical Sciences, Optics & Photonics, Microscopy, New, Two-Photon Microscopy, 2-Photon Microscopy, Photonics


*Licensing
Bruce Toman, Technology Commercialization and Liaison Officertoman@cornell.edu212-746-6187
Country/Region
USA

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