Fluorescence emission difference microscopy (FED) is a method to improve the resolution implemented by the subtraction of two scanning images modulated by two different beams, and the resolution can be enhanced by a factor of 2 at most. For further improvement of the resolving ability of FED, a novel method is proposed in this paper utilizing the technique of imaging scan to realize the higher-resolution imaging, which we called imaging scanning fluorescence emission difference microscopy (ISFED). With a 32-element detector array, the subtraction between imaging results with imaging scanning microscopy and confocal microscopy can achieve higher resolution, including transverse direction and axial direction, and signal noise ratio (SNR) than that in FED microscopy.

© 2016 Optical Society of America

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