Proceeding beyond Diffraction Limit Stochastic Optical Reconstruction Microscopy (STORM)-3D -Abhik Bose, 154033002- CHS 818 For slide and abstract Please.

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Presentation transcript:

Proceeding beyond Diffraction Limit Stochastic Optical Reconstruction Microscopy (STORM)-3D -Abhik Bose, CHS 818 For slide and abstract Please visit

Way-out: Super resolution Microscopy or Nanoscopy: Two approaches 1.Improved optics to achieve sub-wavelength resolution. Techniques include NSOM, SIM, STED etc. 2.Intelligent methodology to achieve super resolution with wide-field setup e.g. Localization Microscopy Pros: High resolution super resolved image Cones: Costly optics, Experimentally challenging. Superresolved image with conventional wide-field setup

Localization Microscopy 1 : FROM:

Stochastic Optical Reconstruction Microscopy 2 Bates, M.; Blosser, T.R.; Zhuang, X. Phys. Rev. Lett. 2005, 94, Rust, M. J.; Bates, M.; Zhuang, X. Nat. Methods 2006, 3, 793

Localisation accuracy with STORM 2

2D STORM Imaging with Cy5-Cy3 Switch 2

Photo switching Mechanism 3,4 Dempsey,. T.; Bates, M.; Kowtoniuk, W. E.; Liu, D. R.; Tsien, R. Y.; Chem. Soc. 2009, 131, 18192–18193.

3D STORM Zhuang Science 2008

Astigmatism (monochromatic) Source: Internet Propagation Source: Optics by Hecht 4 th edition.

Astigmatism and 3D STORM Methodology 6

3D STORM Image 6 Clathrin-coted pits in cell

Re constructed image with 3D STORM 6

Drawbacks Upon invention STORM is a very successful method, but is dependent on storm specific dyes and the dye bleaches permanentely after a few hundreds of cycles. Invention of new dyes will remove this drawbacks. References: 1. Accessed On: :10 pm IST 2.Rust, M. J.; Bates, M.; Zhuang, X. Nat. Methods 2006, 3, Dempsey, G. T.; Bates, M.; Kowtoniuk, W. E.; Liu, D. R.; Tsien, R. Y.; Chem. Soc. 2009, 131, 18192– Bates, M.; Blosser, T.R.; Zhuang, X. Phys. Rev. Lett. 2005, 94, Dempsey, G. T.; Vaughan, J. C.; Chen, K. H.; Bates, M.; Zhuang, X. Nat. Methods 2011, 8, 1027– Huang, B.; Jones, S. A.; Brandenburg, B.; Zhuang, X. Nat. Methods 2008, 5, 1047– 1052.