Fresnel incoherent correlation holographic localization microscopy with enhanced localization accuracy by employing autofocus algorithm

Fresnel incoherent correlation holography as a self-interference holographic technique is employed as three dimensional (3D) localization method of fluorescent microspheres, thus to obtain 3D microscopic imaging. In order to promote the localization accuracy and the reconstructed image quality, autofocus algorithm is introduced to help finding accurate reconstruction distance in the Fresnel Incoherent Correlation Holographic (FINCH) localization microscopic imaging. The performances of autofocus algorithm under different evaluation parameters of reconstruction images are investigated, and the suitable autofocus algorithm are used during hologram reconstruction. The localization algorithm is utilized based on the reconstructed image. The localization accuracy and imaging quality are compared with and without employing the autofocus algorithm. The enhanced localization accuracy is realized in the actual FINCH localization microscopic imaging system.