TY - GEN
T1 - A PSF-based approach to Biplane calibration in 3D super-resolution microscopy
AU - Kirshner, Hagai
AU - Pengo, Thomas
AU - Olivier, Nicolas
AU - Sage, Daniel
AU - Manley, Suliana
AU - Unser, Michael
PY - 2012
Y1 - 2012
N2 - Super-resolution localization microscopy methods such as PALM and STORM have been shown to provide imaging with resolutions up to a few tens of nanometers while using relatively simple setups. Biplane PALM has extended the PALM technique to three-dimensions, by simultaneously using two imaging planes, with different focal depths. A key aspect in achieving good axial localization results is the alignment of the two planes. Currently available approaches assume that misaligned planes only result in scaling and rotation of the PSF pattern. We show in this work that this does not necessarily hold true, especially in the presence of refractive index mismatch between the different optical layers. Instead, we suggest a calibration algorithm that relies on a realistic PSF model and finds the affine transform that relates the two planes with respect to a point source in the object domain. Our calibration algorithm also determines the defocus distance between the planes.
AB - Super-resolution localization microscopy methods such as PALM and STORM have been shown to provide imaging with resolutions up to a few tens of nanometers while using relatively simple setups. Biplane PALM has extended the PALM technique to three-dimensions, by simultaneously using two imaging planes, with different focal depths. A key aspect in achieving good axial localization results is the alignment of the two planes. Currently available approaches assume that misaligned planes only result in scaling and rotation of the PSF pattern. We show in this work that this does not necessarily hold true, especially in the presence of refractive index mismatch between the different optical layers. Instead, we suggest a calibration algorithm that relies on a realistic PSF model and finds the affine transform that relates the two planes with respect to a point source in the object domain. Our calibration algorithm also determines the defocus distance between the planes.
KW - 3D particle localization
KW - point spread function modeling
KW - super-resolution fluorescence microscopy
UR - http://www.scopus.com/inward/record.url?scp=84864862355&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=84864862355&partnerID=8YFLogxK
U2 - 10.1109/ISBI.2012.6235784
DO - 10.1109/ISBI.2012.6235784
M3 - Conference contribution
AN - SCOPUS:84864862355
SN - 9781457718588
T3 - Proceedings - International Symposium on Biomedical Imaging
SP - 1232
EP - 1235
BT - 2012 9th IEEE International Symposium on Biomedical Imaging
T2 - 2012 9th IEEE International Symposium on Biomedical Imaging: From Nano to Macro, ISBI 2012
Y2 - 2 May 2012 through 5 May 2012
ER -