Quantitative orientation-independent differential interference contrast (DIC) microscopy

Michael Shribak; James LaFountain; David Biggs; Shinya Inoué

doi:10.1117/12.709901

19 February 2007 Quantitative orientation-independent differential interference contrast (DIC) microscopy

Michael Shribak, James LaFountain, David Biggs, Shinya Inoué

Proceedings Volume 6441, Imaging, Manipulation, and Analysis of Biomolecules, Cells, and Tissues V; 64411L (2007) https://doi.org/10.1117/12.709901
Event: SPIE BiOS, 2007, San Jose, California, United States

Abstract

We describe a new DIC technique, which records phase gradients within microscopic specimens independently of their orientation. The proposed system allows the generation of images representing the distribution of dry mass (optical path difference) in the specimen. Unlike in other forms of interference microscopes, this approach does not require a narrow illuminating cone. The orientation-independent differential interference contrast (OI-DIC) system can also be combined with orientation-independent polarization (OI-Pol) measurements to yield two complementary images: one showing dry mass distribution (which is proportional to refractive index) and the other showing distribution of birefringence (due to structural or internal anisotropy). With a model specimen used for this work -- living spermatocytes from the crane fly, Nephrotoma suturalis --- the OI-DIC image clearly reveals the detailed shape of the chromosomes while the polarization image quantitatively depicts the distribution of the birefringent microtubules in the spindle, both without any need for staining or other modifications of the cell. We present examples of a pseudo-color combined image incorporating both orientation-independent DIC and polarization images of a spermatocyte at diakinesis and metaphase of meiosis I. Those images provide clear evidence that the proposed technique can reveal fine architecture and molecular organization in live cells without perturbation associated with staining or fluorescent labeling. The phase image was obtained using optics having a numerical aperture 1.4, thus achieving a level of resolution never before achieved with any interference microscope.

Citation Download Citation

Michael Shribak, James LaFountain, David Biggs, and Shinya Inoué "Quantitative orientation-independent differential interference contrast (DIC) microscopy", Proc. SPIE 6441, Imaging, Manipulation, and Analysis of Biomolecules, Cells, and Tissues V, 64411L (19 February 2007); https://doi.org/10.1117/12.709901

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