Abstract

We are proposing to develop the polarized light field microscope for analyzing the architectural dynamics inside living cells, tissues and cell-free model systems. The instrument combines comprehensive polarization analysis and 3D imaging of microscopic specimens in a single snapshot using the new approach of light field imaging. The project includes the development of the hardware, software and algorithms required for capturing polarized light field images and their tomographic reconstruction into 3-dimensional maps of birefringence, diattenuation and polarized fluorescence. The instrument development is guided by several application projects including the remodeling of the extracellular collagen matrix by fibroblast cells, the spatial coordination in the assembly of higher order septin structures inside eukaryotic cells and the origins of chromosome malorientations and their repair in the meiotic spindle of spermatocytes. Information about the technology, including software components and instrument design that are developed under this grant, will be made available for users and developers on an established web portal called OpenPolScope.org.

Public Health Relevance

The microscopy instrumentation developed under this grant will have applications in medical research and clinical settings. Furthermore, the cell biological application projects that will be forthcoming under this grant will have broad implications for our understanding of cell division and cellular remodeling of tissue, which are fundamental processes in health and disease.

  Funding Agency

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
3R01GM114274-03S1
Application #
9527370
Study Section
Program Officer
Sammak, Paul J
Project Start
2015-05-01
Project End
2019-03-31
Budget Start
2017-04-01
Budget End
2018-03-31
Support Year
3
Fiscal Year
2017
Total Cost
Indirect Cost

  Institution

Name
Marine Biological Laboratory
Department
Type
DUNS #
001933779
City
Woods Hole
State
MA
Country
United States
Zip Code
02543

  Publications

Yoshizawa, Takuya; Ali, Rustam; Jiou, Jenny et al. (2018) Nuclear Import Receptor Inhibits Phase Separation of FUS through Binding to Multiple Sites. Cell 173:693-705.e22
Spira, Felix; Cuylen-Haering, Sara; Mehta, Shalin et al. (2017) Cytokinesis in vertebrate cells initiates by contraction of an equatorial actomyosin network composed of randomly oriented filaments. Elife 6:
Gibaud, Thomas; Kaplan, C Nadir; Sharma, Prerna et al. (2017) Achiral symmetry breaking and positive Gaussian modulus lead to scalloped colloidal membranes. Proc Natl Acad Sci U S A 114:E3376-E3384
McQuilken, Molly; Jentzsch, Maximilian S; Verma, Amitabh et al. (2017) Analysis of Septin Reorganization at Cytokinesis Using Polarized Fluorescence Microscopy. Front Cell Dev Biol 5:42
Swaminathan, Vinay; Kalappurakkal, Joseph Mathew; Mehta, Shalin B et al. (2017) Actin retrograde flow actively aligns and orients ligand-engaged integrins in focal adhesions. Proc Natl Acad Sci U S A 114:10648-10653
Nordenfelt, Pontus; Moore, Travis I; Mehta, Shalin B et al. (2017) Direction of actin flow dictates integrin LFA-1 orientation during leukocyte migration. Nat Commun 8:2047
Keikhosravi, Adib; Liu, Yuming; Drifka, Cole et al. (2017) Quantification of collagen organization in histopathology samples using liquid crystal based polarization microscopy. Biomed Opt Express 8:4243-4256
Chandler, Talon; Mehta, Shalin; Shroff, Hari et al. (2017) Single-fluorophore orientation determination with multiview polarized illumination: modeling and microscope design. Opt Express 25:31309-31325
Tani, Tomomi; Shribak, Michael; Oldenbourg, Rudolf (2016) Living Cells and Dynamic Molecules Observed with the Polarized Light Microscope: the Legacy of Shinya Inoué. Biol Bull 231:85-95
Mehta, Shalin B; McQuilken, Molly; La Riviere, Patrick J et al. (2016) Dissection of molecular assembly dynamics by tracking orientation and position of single molecules in live cells. Proc Natl Acad Sci U S A 113:E6352-E6361

Showing the most recent 10 out of 11 publications