Super-Resolution Optical Fluctuation Imaging Using Speckle Illumination

In conventional far-field microscopy, two objects separated closer than approximately half of an emission wavelength cannot be resolved, because of the fundamental limitation known as Abbe's diffraction limit. During the last decade, several super-resolution methods have been developed to overcome the diffraction limit in optical imaging. Among them, super-resolution optical fluctuation imaging (SOFI) developed by Dertinger et al [1], employs the statistical analysis of temporal fluorescence fluctuations induced by blinking phenomena in fluorophores. SOFI is a simple and versatile method for super-resolution imaging. However, due to the uncontrollable blinking of fluorophores, there are some limitations to using SOFI for several applications, including the limitations of available blinking fluorophores for SOFI, a requirement of using a high-speed camera, and a low signal-to-noise ratio. To solve these limitations, we present a new approach combining SOFI with speckle pattern illumination to create illumination-induced optical fluctuation instead of blinking fluctuation of fluorophore.. This technique effectively overcome the limitations of the conventional SOFI since illumination-induced optical fluctuation is possible to control unlike blinking phenomena of fluorophore. And we present the sub-diffraction resolution image using SOFI with speckle illumination.