Rapid confocal microscopy to support surgical procedures

A method for presenting a multimodal image from a confocal microscope on a display device comprising:

• contacting a surface of a biological sample with a solution of a first fluorescent stain that distinguishes a nucleus of a cell, wherein the step of contacting comprises contacting the surface of the biological sample with a solution of acridine orange for a duration selected in a range from about 0.6 milliseconds to about 20 seconds;
• illuminating a spot on the surface of the biological sample with a light beam using the confocal microscope;
• detecting a first emission intensity from the spot in a first range of optical properties, wherein the first emission intensity is intensity from the fluorescent stain;
• detecting a second emission intensity from the spot in a second range of optical properties, wherein the second emission intensity is intensity of unstained reflectance;
• coloring a pixel that corresponds to the spot in an image using a linear combination of the first emission intensity detected from the spot and the second emission intensity detected from the spot, wherein the linear combination includes the first emission intensity times non-zero first factors in a red channel, green channel and blue channel, respectively, to imitate a purple stain of a hematoxylin dye and the second emission intensity times non-zero second factors in the red channel, green channel and blue channel, respectively, to imitate a pink stain of an eosin dye;
• and
• presenting the colored pixels on the display device to approximate the color produced by a conventional histology section for the biological sample at the spot.