The United States of America, As Represented by the Secretary, Department of Health and Human Services

The United States of America As Represented by the Department of Health

Jay R Knutson

Robert S Balaban

Christian A Combs

Gary L Griffiths

Joseph M Schmitt

Alexander Knuttel

Light collectors for fluorescence microscopy include slab light guides having a specimen volume such as a specimen well configured to retain a specimen. The specimen volume is bounded by a surface of the slab light guide that permits laterally propagating fluorescence to enter the slab light guide and propagate toward a detection or imaging system. Typically, the slab light guide has an elliptical perimeter and the sample volume is situated at a first focus of the elliptical perimeter. A reflector such as a conical reflector is situated at the second focus so as to direct the collected fluorescence to exit the slab light guide.

Lightguides to simplify total emission detection for multiphoton microscopy

Composite probes for super resolution optical techniques using super resolution via transiently activated quenchers (STAQ) include a donor moiety and an acceptor moiety joined by a linker, wherein the acceptor moiety, when excited by incident radiation, is excited to a state which, for example, absorbs in the donor emission region, such that the acceptor moiety in its excited state quenches at least a portion of the donor moiety emission. Other transiently activated quenching mechanisms and moieties could accomplish the same task by reducing donor population. Also disclosed are methods for irradiating a selected region of a target material including the composite probe, wherein the composite probe enables improved resolution by point spread function modification and/or nanoscale chemical reactions.

Composite probes and use thereof in super resolution methods

Polarizers and polarizing beam splitter include one or more pairs of axicons that are configured to separate an input beam into a radially polarized component and a tangentially (or azimuthally) polarized component. A second axicon pair can be provided to recombine the tangentially polarized component so as to provide a more uniform beam intensity. The radially polarized component can be reflected or otherwise directed so that one or both the radial and tangential components are available for use.

Azicon beam polarization devices

A multi-photon microscope having an illumination source that transmits an illumination light into a housing having an objective lens arrangement for illuminating a sample disposed outside the housing and directing a first portion of emission light emitted from the sample to a detection system is disclosed. A light collection system is disposed proximate the objective lens arrangement for directing a second portion of emission light in a coaxial relationship with the first portion of emission light to the detection system such that substantially all of the emission light on, around and above the illumination region is detected.

Non-contact total emission detection method and system for multi-photon microscopy

A multi-photon microscope has an illumination source, an objective lens unit arranged in an optical path of the illumination source, a first light collection system arranged to collect a first portion of light emitted from a sample when the sample is illuminated by light from the illumination source, and a second light collection system arranged to collect a second portion of light emitted from the sample when the sample is illuminated by light from the illumination source. The first portion of light when collected by the first light collection system and the second portion of light when collected by the second light collection system, together provide a means of collecting as much light from as many angles as possible emanating from an emitting point source. This collection scheme has the potential to approach the total emission collection of light from an emitting point source depending on the optical properties of the sample being imaged.

Wide-area fluorescence detection system for multi-photon microscopy

An apparatus and method for using frequency modulated diffusive photon-density waves propagating through a turbid medium such as tissue for detecting optical properties of objects and for imaging objects within the turbid medium. Frequency modulation enables real time differential measurement of the interaction of different colors with the object when the different colors are chosen such that they react the same with the medium but differently with the object. The method greatly simplifies optical imaging and increases the amount of information which may be obtained about the object.

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Kensington, MD, United States of America

Jay R Knutson