Virtual spatially modulated illumination microscopy predicts nanometer precision of axial distance measurment

A. V. Failla; Christoph G. Cremer

doi:10.1117/12.426779

10 May 2001 Virtual spatially modulated illumination microscopy predicts nanometer precision of axial distance measurment

A. V. Failla, Christoph G. Cremer

Author Affiliations +

Proceedings Volume 4260, Optical Diagnostics of Living Cells IV; (2001) https://doi.org/10.1117/12.426779
Event: BiOS 2001 The International Symposium on Biomedical Optics, 2001, San Jose, CA, United States

Abstract

Far field optical light microscopy with its unique capability for contactless, non destructive imaging inside thick transparent specimen such as cell nuclei has contributed widely to the present knowledge of the three- dimensional (3D-) architecture of the interphase nucleus. A serious drawback, however, is the limited optical resolution. A recently introduced light microscopical approach, Spectral Precision Distance Microscopy (SPDM) allows the measurement of distances between point-like fluorescent objects of different spectral signature far below the optical resolution criterion as defined by the Full Width at Half Maximum (FWHM) of the point spread function (PSF). Here, an aspect of the theoretical limits of this method was studied by virtual microscopy. The precision of the axial distance measurements was studied, taking into account photon statistics and image analysis. The results indicate that even under low fluorescence intensity conditions typical for biological structure research, a precision of distance measurements in the nanometer range can be determined.

Citation Download Citation

A. V. Failla and Christoph G. Cremer "Virtual spatially modulated illumination microscopy predicts nanometer precision of axial distance measurment", Proc. SPIE 4260, Optical Diagnostics of Living Cells IV, (10 May 2001); https://doi.org/10.1117/12.426779

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