Abstract
We present a de novo design of an objective for use in multi-photon (MPM) and second harmonic generation (SHG) microscopy. This objective was designed to have a large field of view (FOV), while maintaining a moderate numerical aperture (NA) and relative straight forward construction. A dichroic beam splitter was incorporated within the objective itself allowing for an increase in the front aperture of the objective and corresponding enhancement of the solid angle of collected emission by an order of magnitude over existing designs.
Highlights
Multiphoton microscopy [1, 2] has developed into a standard tool for the life scientist with far reaching applications ranging from basic cell biology to imaging physiology and disease progression in live animals
As the technique has advanced there has been a corresponding development of new objective lenses for use in multiphoton microscopy
By incorporating a dichroic element into the objective itself, the need to collimate this light in order to send it to a distant detector can be relaxed. 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 numerical apertures (NA) alone
Summary
Multiphoton microscopy [1, 2] has developed into a standard tool for the life scientist with far reaching applications ranging from basic cell biology to imaging physiology and disease progression in live animals. We set a design threshold of no more than 25 mm for the diameter of this stop, which we can accommodate in our scanning systems using a scan lens/tube lens combination that produces a ~12x increase in the laser beam diameter to provide a nearly overfilled back aperture This reduces our scan angle, this lens design achieves a large FOV at relatively small field angles. By incorporating a dichroic element into the objective itself, the need to collimate this light in order to send it to a distant detector can be relaxed 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. With the current trend of pre-compensation systems being incorporated into commercial Ti:Sapphire lasers (e.g. Spectra Physics DeepSee) it should be possible to deliver transform limited pulses through it
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