Adaptive holographic region of interest illumination with oblique angles for use in single molecule localization microscopy

Alexander Jügler; Jan Becker; Ronny Förster; Rainer Heintzmann

doi:10.1117/12.2525253

22 July 2019 Adaptive holographic region of interest illumination with oblique angles for use in single molecule localization microscopy

Alexander Jügler, Jan Becker, Ronny Förster, Rainer Heintzmann

Author Affiliations +

Proceedings Volume 11076, Advances in Microscopic Imaging II; 1107602 (2019) https://doi.org/10.1117/12.2525253
Event: European Conferences on Biomedical Optics, 2019, Munich, Germany

Abstract

To improve the localization accuracy and to reduce the overall photo-damage in Single Molecule Localization Microscopy (SMLM), we introduce a holography based Region-of-Interest (ROI) illumination with oblique angles. The ROI illumination is realized in two different application modes which offer different advantages. Both modes (A,B) are implemented within the same setup by modifying the illumination light using a phase-only spatial light modulator (SLM) twice. This allows to adaptively modify the size and the (excitation) angle of the required ROI illumination, resulting in reduced out-of focus signal and less overall phototoxicity. Mode A generates a nearly speckle free, circular ROI which can be obtained instantly (no iterative algorithm needed). Mode B allows to realize an arbitrarily shaped ROI but comes at the cost of a higher presence of speckle structures as well as in an increased calculation time of the holograms. In both cases illumination angles up to 60° with high NA objectives are realizable, enabling the effective selection of specific parts of a biological probe to be illuminated.

Citation Download Citation

Alexander Jügler, Jan Becker, Ronny Förster, and Rainer Heintzmann "Adaptive holographic region of interest illumination with oblique angles for use in single molecule localization microscopy", Proc. SPIE 11076, Advances in Microscopic Imaging II, 1107602 (22 July 2019); https://doi.org/10.1117/12.2525253

ACCESS THE FULL ARTICLE

INSTITUTIONAL
Select your institution to access the SPIE Digital Library.

SELECT YOUR INSTITUTION

PERSONAL
Sign in with your SPIE account to access your personal subscriptions or to use specific features such as save to my library, sign up for alerts, save searches, etc.

PERSONAL SIGN IN

No SPIE Account? Create one

PURCHASE THIS CONTENT

SUBSCRIBE TO DIGITAL LIBRARY

50 downloads per 1-year subscription

Members: $195

Non-members: $335 ADD TO CART

25 downloads per 1 - year subscription

Members: $145

Non-members: $250 ADD TO CART

PURCHASE SINGLE ARTICLE

Includes PDF, HTML & Video, when available

Members: $17.00

Non-members: $21.00 ADD TO CART

PROCEEDINGS
3 PAGES

DOWNLOAD PAPER SAVE TO MY LIBRARY

GET CITATION

RIGHTS & PERMISSIONS

Get copyright permission Get copyright permission on Copyright Marketplace

KEYWORDS

Holography

Microscopy

Spatial light modulators

Molecules

Holograms

Geometrical optics

In vivo imaging

Show All Keywords

Keywords/Phrases

Search In:

Publication Years