FALCON: fast and unbiased reconstruction of high-density super-resolution microscopy data.
Super resolution microscopy such as STORM and (F)PALM is now a well known method for biological studies at the nanometer scale. However, conventional imaging schemes based on sparse activation of photo-switchable fluorescent probes have inherently slow temporal resolution which is a serious limitation when investigating live-cell dynamics. Here, we present an algorithm for high-density super-resolution microscopy which combines a sparsity-promoting formulation with a Taylor series approximation of the PSF. Our algorithm is designed to provide unbiased localization on continuous space and high recall rates for high-density imaging, and to have orders-of-magnitude shorter run times compared to previous high-density algorithms. We validated our algorithm on both simulated and experimental data, and demonstrated live-cell imaging with temporal resolution of 2.5 seconds by recovering fast ER dynamics.
Department of Bio and Brain Engineering, KAIST, Daejeon, Republic of Korea.
Junhong
J
Min
1] Institute of Microengineering, EPFL, Switzerland [2].
Cédric
C
Vonesch
1] Institute of Microengineering, EPFL, Switzerland [2].
Hagai
H
Kirshner
1] Institute of the Physics of Biological Systems, EPFL, Switzerland [2].
Lina
L
Carlini
1] Institute of the Physics of Biological Systems, EPFL, Switzerland [2] Department of Physics, King's College London, UK.
Nicolas
N
Olivier
Institute of the Physics of Biological Systems, EPFL, Switzerland.
Seamus
S
Holden
Institute of the Physics of Biological Systems, EPFL, Switzerland.
Suliana
S
Manley
Department of Bio and Brain Engineering, KAIST, Daejeon, Republic of Korea.
Jong Chul
JC
Ye
Institute of Microengineering, EPFL, Switzerland.
Michael
M
Unser
D000465:Algorithms; D000818:Animals; D019556:COS Cells; D002232:Carbocyanines / Q000737:chemistry; D002460:Cell Line; D002522:Chlorocebus aethiops; D004721:Endoplasmic Reticulum / Q000378:metabolism; D005456:Fluorescent Dyes / Q000737:chemistry; D006801:Humans; D008856:Microscopy, Fluorescence; D008870:Microtubules / Q000737:chemistry / Q000378:metabolism; D059629:Signal-To-Noise Ratio
C569686:Alexa Fluor 647; D002232:Carbocyanines; D005456:Fluorescent Dyes