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 (5b, 5c) 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 (5b, 5c) 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.