A system to segment an image, comprising: a high content imaging system for obtaining a microscopy image and/or a series of microscopy images of a biological sample, wherein the high content imaging system includes an image capture device; an image acquisition module that is adapted to receive an image captured by the image capture device; a coarse object detection module that is adapted to develop a coarse segmentation image, wherein each pixel of the coarse segmentation image is associated with a corresponding pixel in the captured image and is identified as one of an object pixel and a background pixel; a marker identification module that is adapted to select at least one marker pixel from the pixels of the coarse segmentation image, wherein each marker pixel is one of a contiguous group of object pixels in the coarse segmentation image that is furthest from a background pixel relative to neighboring pixels of the group, an object splitting module adapted to associate a pixel of a segmented image corresponding to an object pixel with a marker pixel, and a human interface module; wherein the object splitting module comprises a plurality of processors adapted to operate in parallel and associates the pixel of the segmented image with the marker pixel by setting the color value of the object pixel to the color value associated with the marker pixel if a distance based metric between the object pixel and the marker pixel is less than the distance based metric between the object pixel and any other marker pixel in the coarse segmentation image, and wherein the human interface module is adapted to generate a masked image from the captured image by: generating a masked image with same dimensions as the captured image; initializing the color value of each pixel of the masked image to a predetermined background color; selecting a pixel of the masked image and a corresponding pixel of the segmented image; determining if the color value of the corresponding pixel in the segmented image is identical to the color value of a marker pixel, wherein a particular cell size is associated with the color value associated with the marker pixel, and setting the color value of the pixel of the masked image identical to a color value of a corresponding pixel of the captured image, if the color value of the corresponding pixel in the segmented image is identical to the color value of the marker pixel. a high content imaging system for obtaining a microscopy image and/or a series of microscopy images of a biological sample, wherein the high content imaging system includes an image capture device; an image acquisition module that is adapted to receive an image captured by the image capture device; a coarse object detection module that is adapted to develop a coarse segmentation image, wherein each pixel of the coarse segmentation image is associated with a corresponding pixel in the captured image and is identified as one of an object pixel and a background pixel; a marker identification module that is adapted to select at least one marker pixel from the pixels of the coarse segmentation image, wherein each marker pixel is one of a contiguous group of object pixels in the coarse segmentation image that is furthest from a background pixel relative to neighboring pixels of the group, an object splitting module adapted to associate a pixel of a segmented image corresponding to an object pixel with a marker pixel, and a human interface module; wherein the object splitting module comprises a plurality of processors adapted to operate in parallel and associates the pixel of the segmented image with the marker pixel by setting the color value of the object pixel to the color value associated with the marker pixel if a distance based metric between the object pixel and the marker pixel is less than the distance based metric between the object pixel and any other marker pixel in the coarse segmentation image, and wherein the human interface module is adapted to generate a masked image from the captured image by: generating a masked image with same dimensions as the captured image; initializing the color value of each pixel of the masked image to a predetermined background color; selecting a pixel of the masked image and a corresponding pixel of the segmented image; determining if the color value of the corresponding pixel in the segmented image is identical to the color value of a marker pixel, wherein a particular cell size is associated with the color value associated with the marker pixel, and setting the color value of the pixel of the masked image identical to a color value of a corresponding pixel of the captured image, if the color value of the corresponding pixel in the segmented image is identical to the color value of the marker pixel. generating a masked image with same dimensions as the captured image; initializing the color value of each pixel of the masked image to a predetermined background color; selecting a pixel of the masked image and a corresponding pixel of the segmented image; determining if the color value of the corresponding pixel in the segmented image is identical to the color value of a marker pixel, wherein a particular cell size is associated with the color value associated with the marker pixel, and setting the color value of the pixel of the masked image identical to a color value of a corresponding pixel of the captured image, if the color value of the corresponding pixel in the segmented image is identical to the color value of the marker pixel.