Methods for microscopy with ultraviolet surface excitation (MUSE) imaging
AssigneeCase Western Reserve University
InventorsYehe Liu, Michael Jenkins
Life of this patent
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Abstract
Described herein are methods for imaging a fluorescent bioassay (including a substrate, such as dispersed cellular sample, exposed to one or more exogenous fluorophore and/or fluorescent probe that accumulate in a structure of interest). The bioassay can be excited with Type-C ultraviolet (UVC) light produced by one or more light emitting diode (LED). The UVC can have a center wavelength that causes emission by the fluorescent bioassay. A digital optical device can collect a signal emitted from the fluorescent bioassay in response to the excitation. The methods relate in particular to Microscopy with Ultraviolet Surface Excitation (MUSE) imaging.
Claims20
A method comprising:
- exposing a dispersed cellular sample to one or more exogenous fluorophore and/or fluorescent probe that accumulate in a structure of interest;
- positioning the dispersed cellular sample in contact with a top face of an optical window of an external accessory, wherein the external accessory comprises the optical window and one or more light emitting diodes (LED) configured to provide Type-C ultraviolet (UVC) light;
- positioning the external accessory in focus with a microscope having one or more digital image sensor;
- exciting the one or more exogenous fluorophore and/or fluorescent probe within the sample with the UVC light, produced by the one or more LED, having a center emission wavelength that causes emission by the one or more exogenous fluorophore and/or fluorescent probe, wherein the UVC light transmits through one or more side faces of the optical window and the optical window delivers the UVC light uniformly to a surface of the dispersed cellular sample in contact with the top face of the optical window via frustrated total internal reflection;
- and
- collecting, with the microscope having the one or more digital image sensor, emitted light from excited ones of the one or more exogenous fluorophores and/or fluorescent probe that creates an image comprising the structure of interest in the sample, wherein the emitted light emits out a bottom face of the optical window to the microscope having the one or more digital image sensor.
A method comprising:
- obtaining a fluorescent bioassay;
- positioning the bioassay in contact with a top face of an optical window of an external accessory, wherein the external accessory comprises the optical window and one or more light emitting diodes (LED) configured to provide Type-C ultraviolet (UVC) light;
- positioning the external accessory in focus with a digital optical detecting device;
- exciting the bioassay with the UVC light, produced by the one or more LED, having a center emission wavelength that causes emission by the fluorescent bioassay, wherein the UVC light transmits through one or more side faces of the optical window and the optical window delivers the UVC light uniformly to a surface of the bioassay in contact with the top face of the optical window via frustrated total internal reflection;
- and
- collecting, with the digital optical detecting device, a signal emitted from the fluorescent bioassay through a bottom face of the optical window towards the digital optical detecting device.
Dependencies are inferred from the claim text.