Abstract: This disclosure includes an imaging system that is configured to image in parallel multiple focal planes in a sample uniquely onto its corresponding detector while simultaneously reducing blur on adjacent image planes. For example, the focal planes can be staggered such that fluorescence detected by a detector for one of the focal planes is not detected, or is detected with significantly reduced intensity, by a detector for another focal plane. This enables the imaging system to increase the volumetric image acquisition rate without requiring a stronger fluorescence signal. Additionally or alternatively, the imaging system may be operated at a slower volumetric image acquisition rate (e.g., that of a conventional microscope) while providing longer exposure times with lower excitation power. This may reduce or delay photo-bleaching (e.g., a photochemical alteration of the dye that causes it to no longer be able to fluoresce), thereby extending the useful life of the sample.

Abstract: An imaging system configured to transform a lateral scan motion into axial refocusing, thus enabling aberration free remote focusing. In one embodiment, the imaging system provides aberration free remote focusing by causing a beam of light to be scanned over a stationary mirror having a variable height in the scan direction, thereby causing the beam of light to be defocused in an axial direction in the return direction (e.g., the direction of propagation for the reflected beam of light). By configuring the back-reflected light resulting from the lateral scan component as described herein, a pure axial scan motion may be obtained at the rate of the employed (lateral) scanning technology (e.g., a GSM). This capability allows the imaging system of the present inventive concept to leverage rapid lateral scan technologies to produce rapid axial displacement of the axial focus of the beam of light.

Abstract: This disclosure includes an imaging system that is configured to image in parallel multiple focal planes in a sample uniquely onto its corresponding detector while simultaneously reducing blur on adjacent image planes. For example, the focal planes can be staggered such that fluorescence detected by a detector for one of the focal planes is not detected, or is detected with significantly reduced intensity, by a detector for another focal plane. This enables the imaging system to increase the volumetric image acquisition rate without requiring a stronger fluorescence signal. Additionally or alternatively, the imaging system may be operated at a slower volumetric image acquisition rate (e.g., that of a conventional microscope) while providing longer exposure times with lower excitation power. This may reduce or delay photo-bleaching (e.g., a photochemical alteration of the dye that causes it to no longer be able to fluoresce), thereby extending the useful life of the sample.

Abstract: Light-sheet fluorescence microscopy (LSFM) affords highly parallelized 3D imaging with optical sectioning capability and minimal light exposure. However, using Gaussian beams for light-sheet generation results in a trade-off between beam waist thickness and the area over which the beam can approximate a light-sheet. Novel techniques for LSFM are disclosed that uses extended focusing and/or laser line focuses to produce divergence free light-sheets with near diffraction-limited resolution and uniform intensity distribution.

Abstract: Systems, methods, and computer-readable storage media are disclosed for providing a structured total internal reflection fluorescence (sTIRF) imaging system providing improved out of focus blur rejection and improved image contrast. The sTIRF imaging system may be configured to illuminate a sample using two beams of light (e.g., a primary beam of light and an interfering beam of light). The interfering beam of light may be configured to create interference with respect to the primary beam of light. The sTIRF imaging system may be configured to capture a plurality of intermediate images of the sample during the illuminating, and to generate a final image of the sample based on the plurality of intermediate images. The interference caused by the two beams of light may enable the sTIRF imaging system to reject out of focus blur based on detection of whether fluorescence emissions from the sample being imaged fluctuate or remain static.

Abstract: This disclosure includes an imaging system that is configured to image in parallel multiple focal planes in a sample uniquely onto its corresponding detector while simultaneously reducing blur on adjacent image planes. For example, the focal planes can be staggered such that fluorescence detected by a detector for one of the focal planes is not detected, or is detected with significantly reduced intensity, by a detector for another focal plane. This enables the imaging system to increase the volumetric image acquisition rate without requiring a stronger fluorescence signal. Additionally or alternatively, the imaging system may be operated at a slower volumetric image acquisition rate (e.g., that of a conventional microscope) while providing longer exposure times with lower excitation power. This may reduce or delay photo-bleaching (e.g., a photochemical alteration of the dye that causes it to no longer be able to fluoresce), thereby extending the useful life of the sample.

Abstract: This disclosure includes an imaging system that is configured to image in parallel multiple focal planes in a sample uniquely onto its corresponding detector while simultaneously reducing blur on adjacent image planes. For example, the focal planes can be staggered such that fluorescence detected by a detector for one of the focal planes is not detected, or is detected with significantly reduced intensity, by a detector for another focal plane. This enables the imaging system to increase the volumetric image acquisition rate without requiring a stronger fluorescence signal. Additionally or alternatively, the imaging system may be operated at a slower volumetric image acquisition rate (e.g., that of a conventional microscope) while providing longer exposure times with lower excitation power. This may reduce or delay photo-bleaching (e.g., a photochemical alteration of the dye that causes it to no longer be able to fluoresce), thereby extending the useful life of the sample.

Abstract: Light-sheet fluorescence microscopy (LSFM) affords highly parallelized 3D imaging with optical sectioning capability and minimal light exposure. However, using Gaussian beams for light-sheet generation results in a trade-off between beam waist thickness and the area over which the beam can approximate a light-sheet. Novel techniques for LSFM are disclosed that uses extended focusing and/or laser line focuses to produce divergence free light-sheets with near diffraction-limited resolution and uniform intensity distribution.

Abstract: Systems, methods, and computer-readable storage media are disclosed for providing a structured total internal reflection fluorescence (sTIRF) imaging system providing improved out of focus blur rejection and improved image contrast. The sTIRF imaging system may be configured to illuminate a sample using two beams of light (e.g., a primary beam of light and an interfering beam of light). The interfering beam of light may be configured to create interference with respect to the primary beam of light. The sTIRF imaging system may be configured to capture a plurality of intermediate images of the sample during the illuminating, and to generate a final image of the sample based on the plurality of intermediate images. The interference caused by the two beams of light may enable the sTIRF imaging system to reject out of focus blur based on detection of whether fluorescence emissions from the sample being imaged fluctuate or remain static.