A fluorescence imaging system, comprising: a sample platform ; and an illumination and detection system, including: an illumination system that provides illumination light from an illumination source to a targeted area on the sample platform , wherein the targeted area includes a fluorescent material, and wherein the illumination light includes light having an excitation wavelength of the fluorescent material; a sensor adapted to detect light and having an array of sensing locations; and collection optics arranged and configured to project light emanating from the sample platform onto the sensor , wherein fluorescent light from the targeted area includes fluorescence emissions from the fluorescent material and is projected onto a first portion of the sensor comprising a first plurality of the sensing locations and background light from a region proximal to the targeted area on the platform is projected onto a second portion of the sensor comprising a second plurality of the sensing locations, and characterized in that a second signal detected by the second portion of the sensor is subtracted from a first signal detected by the first portion of the sensor to produce a background-reduced signal, where an illumination pattern in the targeted area is a point, a number of points, a line, or number of lines and the sample platform and/or the system are scanned in one or both directions to build up an image of larger area. a sample platform ; and an illumination and detection system, including: an illumination system that provides illumination light from an illumination source to a targeted area on the sample platform , wherein the targeted area includes a fluorescent material, and wherein the illumination light includes light having an excitation wavelength of the fluorescent material; a sensor adapted to detect light and having an array of sensing locations; and collection optics arranged and configured to project light emanating from the sample platform onto the sensor , an illumination system that provides illumination light from an illumination source to a targeted area on the sample platform , wherein the targeted area includes a fluorescent material, and wherein the illumination light includes light having an excitation wavelength of the fluorescent material; a sensor adapted to detect light and having an array of sensing locations; and collection optics arranged and configured to project light emanating from the sample platform onto the sensor , wherein fluorescent light from the targeted area includes fluorescence emissions from the fluorescent material and is projected onto a first portion of the sensor comprising a first plurality of the sensing locations and background light from a region proximal to the targeted area on the platform is projected onto a second portion of the sensor comprising a second plurality of the sensing locations, and characterized in that a second signal detected by the second portion of the sensor is subtracted from a first signal detected by the first portion of the sensor to produce a background-reduced signal, where an illumination pattern in the targeted area is a point, a number of points, a line, or number of lines and the sample platform and/or the system are scanned in one or both directions to build up an image of larger area. <EOS>