Spectral unmixing

• Main Authors: Endsley, Eric William, Bierdz, Paul Joseph, Holtz, Nevine
• Format: Patent
• Language: English
• Published: 28.07.2020
• Subjects: CALCULATING; COMPUTING; COUNTING; IMAGE DATA PROCESSING OR GENERATION, IN GENERAL; INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIRCHEMICAL OR PHYSICAL PROPERTIES; MEASURING; PHYSICS; TESTING

Systems and methods are provided for microscopically and fluorescently imaging cell-bearing biological samples or other samples of interest. A microscope objective or other optical elements that exhibits chromatic aberration can be used to obtain images of fluorophores or other contrast agents at different wavelengths. The obtained images are then used to correct each other, e.g., to remove artifacts in an image of a shorter-wavelength fluorophore that are caused by cross-talk from a longer-wavelength fluorophore. A longer-wavelength image, taken at a focal distance corresponding to the shorter-wavelength fluorophore, is taken and used to subtract the activity of the longer-wavelength fluorophore in the shorter-wavelength image. The longer-wavelength image may be taken using a microscope set to the shorter-wavelength focal distance. Alternatively, the longer-wavelength image may be simulated by applying a blurring filter or other methods to a longer-wavelength image taken when the microscope is set to the longer-wavelength focal distance.