Optogenetics microscope
First Claim
Patent Images
1. A near-total internal reflection fluorescence microscope comprising:
- a stage comprising an object region configured to support a sample;
an illumination subsystem comprising a prism disposed beneath the object region, the prism comprising an upper surface proximal to the object region, a lower surface distal to the object region, and at least one side surface adjoining the upper surface and the lower surface, and a light source operable to transmit light onto the at least one side surface of the prism without achieving total internal reflection when the sample is on the stage, causing the prism to illuminate the sample from beneath by near-total internal reflection; and
an imaging subsystem comprising an objective lens unit disposed beneath the prism and an image capture device operable to receive an image passed through the objective lens unit from the object region.
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Abstract
The invention provides an open-stage near-TIRF microscope in which all of the optical components are positioned underneath the sample, allowing for physical access to, and control over the environment of, the sample. The microscope can be used to image cells expressing fluorescent voltage indicators. Since the TIRF components do not interfere with the sample, living cells can be studied using a microscope of the invention. Where a sample includes electrically active cells expressing fluorescent voltage indicators, the microscope can be used to view voltage changes in, and thus the electrical activity of, those cells.
63 Citations
20 Claims
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1. A near-total internal reflection fluorescence microscope comprising:
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a stage comprising an object region configured to support a sample; an illumination subsystem comprising a prism disposed beneath the object region, the prism comprising an upper surface proximal to the object region, a lower surface distal to the object region, and at least one side surface adjoining the upper surface and the lower surface, and a light source operable to transmit light onto the at least one side surface of the prism without achieving total internal reflection when the sample is on the stage, causing the prism to illuminate the sample from beneath by near-total internal reflection; and an imaging subsystem comprising an objective lens unit disposed beneath the prism and an image capture device operable to receive an image passed through the objective lens unit from the object region. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20)
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Specification