Improve resolution and go 3D
The quality of the Point Spread Function (PSF) is crucial in single molecule localization super resolution methods (SMLM). By correcting both setup and sample induced aberrations, adaptive optics optimizes the shape of the PSF and this way improves the number of detected photons. This directly enhances the localization precision in these methods. In addition to that, by applying the controllable amount of perfect astigmatism, adaptive optics enable the best 3D imaging capability. This pure astigmatism superimposed onto corrected PSF delivers unprecedented accuracy of single molecule localization in all three dimensions. For an easy implementation of adaptive optics into microscopy setup, Imagine Optic developed a plug and play solution MicAO 3DSR.
Combined two-color 3D PALM and dSTORM image of mEos2-Centrin1 (orange) and Alexa 647 labeled Cep164 (blue). The data is obtained using MicAO 3DSR. Image is taken from Clouvel et al (2013) Proc SPIE, 8590, 85 900Z+.
PALM/STORM and SPT
M. Marbouty, A. Le Gall, D. I. Cattoni, A. Cournac, A. Koh, J-B. Fiche, J. Mozziconacci, H. Murray, R. Koszul, M. Nollmann (2015) “Condensin- and Replication-Mediated Bacterial Chromosome Folding and Origin Condensation Revealed by Hi-C and Super-resolution Imaging.” Mol. Cell, 59, 588-602.
K. F. Tehrani, J. Xu, Y. Zhang, P. Shen, P. Kner (2015) “Adaptive optics stochastic optical reconstruction microscopy (AO-STORM) using a genetic algorithm.” Opt. Express, 23, 13677-13692.
M. Petkovic, A. Jemaiel, F. Daste, C. G. Specht, I. Izeddin, D. Vorkel, J-M. Verbavatz, X. Darzacq, A. Triller, K. H. Pfenninger, D. Tareste, C. L. Jackson, T. Galli (2014) “The SNARE Sec22b has a non-fusogenic function in plasma membrane expansion.” Nat. Cell Biol., 16, 434-444.
B. Hajj, M. El Beheiry, I. Izeddin, X. Darzacq, M. Dahan (2014) “Accessing the third dimension in localization-based super-resolution microscopy.” Phys. Chem. Chem. Phys., 16, 16340.
V. Récamier, I. Izeddin, L. Bosanac, M. Dahan, F. Proux, X. Darzacq (2014) “Single cell correlation fractal dimension of chromatin: A framework to interpret 3D single molecule super-resolution.” Nucleus, 5.
C. G. Specht, I. Izeddin, P. C. Rodriguez, M. El Beheiry, P. Rostaing, X. Darzacq, M. Dahan, A. Triller. (2013) “Quantitative nanoscopy of inhibitory synapses: Counting gephyrin molecules and receptor binding sites.” Neuron, 79, 308–321.
G. Clouvel, A. Jasaitis, J. Sillibourne, I. Izeddin, M. El Beheiry, X. Levecq, M. Dahan, M. Bornens, X. Darzacq. (2013) “Dual-color 3D PALM/dSTORM imaging of centrosomal proteins using MicAO 3DSR.” Proc. SPIE, 8590, 85 900Z+.
I. Izeddin, M. El Beheiry, J. Andilla, D. Ciepielewski, X. Darzacq, M. Dahan. (2012) “PSF shaping using adaptive optics for three-dimensional single-molecule super-resolution imaging and tracking.” Opt. Express, 20, 4957–4967.