Using temporal correlations of fluorescence intensity to improve single molecule localization microscopy

  Shachar Schidorsky  ,  Shachar Schidorsky [2]  ,  Xiyu Yi [1]  ,  Yair Razvag [2]  ,  Julia Sajman [2]  ,  Yonatan Golan [2]  ,  Shimon Weiss [1]  ,  Eilon Sherman [2]  
[1] Department of Chemistry & Biochemistry, UCLA, CA
[2] Racah Institute of Physics, The Hebrew University, Jerusalem

Single molecule localization microscopy (SMLM) enable the localization of individual molecules within intact cells with a resolution of ~20nm. However, SMLM requires longer acquisition time compared to diffraction limited microscopy and is often challenging under low signal to background (SBR) conditions. Here, we utilized temporal intensity fluctuations of fluorophores to efficiently reject background in acquired SMLM movies of various tagged proteins in fixed and live T cells. Our approach could be used to automatically enhance SMLM imaging under detrimental imaging conditions.