# Orientation Independent DIC and Polarization Microscopy

> **NIH NIH R01** · MARINE BIOLOGICAL LABORATORY · 2020 · $472,800

## Abstract

Project Summary
This project aims to develop and apply new quantitative differential interference contrast and polarized light
microscope techniques, both of whose contrast is independent of specimen orientation. During the previous
grant period we built the orientation-independent differential interference contrast (OI-DIC) microscope, which
rapidly changes shear directions without any mechanically moved components. The OI-DIC technique, which
we have developed, provides the highest quality of optical path length (dry mass) map of thin optical section of
unstained and stained specimens with lateral resolution ~250-300nm, axial discrimination depth ~100nm, and
optical path length sensitivity ~0.5nm at wavelength 546nm. We believe that images with such high level of
resolution cannot be produced by any other currently available interference and phase microscopy techniques.
We also built the polychromatic polarized light microscope (polscope), which employs new principle of
generating interference color and produces a color image of the birefringent structures with retardances of
several nm, which was not possible before. The hue of the structure indicates its slow axis orientation, and the
brightness of the structure is proportional to its retardance. We will engineer the improved high-resolution OI-
DIC with and exploit two principally new OI-DIC approaches. The new OI-DIC will be combined with the
orientation-independent polarization (LC-polscope) and confocal fluorescence techniques. The improved OI-
DIC and the combined setup will be used to study the architectural dynamics of live biological specimens, with
emphasis on events associated with mitosis and meiosis. We will build an instantaneous OI-DIC, which will
simultaneously capture 4 raw DIC images with the orthogonal shear directions. The instantaneous OI-DIC
techniques will provide the best temporal resolution and allow the elimination of artifacts caused by movements
of the cytosol and organelles. We will develop OI-DIC technique to restore the 3D distribution of both dry mass
(phase-related information) and refractive index. With this technique, one will be able to determine the dry
mass of chromosomes before division and observe its change during the division process. The new high-
sensitive polychromatic polscope will visualize birefringent structures with retardances less than 1nm. The
polychromatic polscope will be combined with phase contrast and dark field techniques, so that image
brightness displays dry mass distribution and the color depicts molecular orientation. These combined
techniques will be used to study metaphase of meiosis I, and analyze the correspondence between structural
signatures in different live organisms, as well as their genetic background. We will create a quantitative
polychromatic polscope, which will provide a two-dimensional distribution both the specimen’s retardance and
slow axis orientation. The quantitative polychromatic polscope will be employed to ...

## Key facts

- **NIH application ID:** 9985864
- **Project number:** 5R01GM101701-11
- **Recipient organization:** MARINE BIOLOGICAL LABORATORY
- **Principal Investigator:** MICHAEL SHRIBAK
- **Activity code:** R01 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2020
- **Award amount:** $472,800
- **Award type:** 5
- **Project period:** 2005-12-01 → 2024-07-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/9985864

## Citation

> US National Institutes of Health, RePORTER application 9985864, Orientation Independent DIC and Polarization Microscopy (5R01GM101701-11). Retrieved via AI Analytics 2026-05-23 from https://api.ai-analytics.org/grant/nih/9985864. Licensed CC0.

---

*[NIH grants dataset](/datasets/nih-grants) · CC0 1.0*