Spatial light interference tomography
View Patent ↗Methods and a computer readable medium for deriving a quantitative phase contrast tomographic image of a specimen. The specimen is illuminated and a focus of the illuminating light is scanned to a plurality of depths within the specimen. Light transmitted through the specimen is spatially Fourier transformed at each of the plurality of depths to form a spatially transformed image, and at least one of the phase and amplitude of a plurality of spatial frequency components of the spatially transformed image is spatially modulated. An intensity image of the specimen plane as modulated with respect to spatial frequency components is detected and deconvolved at a plurality of spatially modulated instances to obtain a three-dimensional phase representation of the specimen at each of the plurality of distances relative to a fiducial plane.
1. A method for deriving a quantitative phase contrast tomographic image of a specimen, the method comprising steps of:
a. illuminating the specimen with illuminating light characterized by a coherence length substantially shorter than a dimension of a feature of the specimen to be resolved;
b. scanning a focus of the illuminating light to a plurality of depths within the specimen;
c. spatially Fourier transforming light transmitted through the specimen at each of the plurality of depths to form a spatially Fourier transformed image;
d. spatially modulating at least one of the phase and amplitude of a plurality of spatial frequency components of the spatially Fourier transformed image;
e. detecting an intensity image of the specimen plane as modulated with respect to spatial frequency components and as interfered with an unscattered component of the illuminating light;
f. deconvolving the intensity image at a plurality of spatially modulated instances to obtain a three-dimensionally registered phase representation of light scattered by the specimen at each of the plurality of distances relative to a fiducial plane; and
g. solving for a quantitative phase contrast tomographic image of the specimen by an inverse scattering solution, by iterative search for a point spread function entirely in a spectral domain and subject to a regularization parameter, based on a linear forward model of scattering through an imaging system.
2. A method in accordance with claim 1 , further comprising deconvolving a modeled point spread function from the three-dimensional phase representation iteratively in the spectral domain.
3. A method in accordance with claim 2 , further comprising a preliminary step of imaging point scatterers to obtain the modeled point spread function.
4. A non-transitory computer readable medium for use on a computer system for deriving a quantitative phase contrast tomographic image of a specimen, the non-transitory computer readable medium having computer readable program code thereon, the computer readable program code comprising:
a. a computer code module for scanning a focus of illuminating light to a plurality of depths within the specimen;
b. a computer code module for spatially Fourier transforming light transmitted through the specimen at each of the plurality of depths to form a spatially transformed image;
c. a computer code module for spatially modulating at least one of the phase and amplitude of a plurality of spatial frequency components of the spatially transformed image;
d. a computer code module for deconvolving a detected intensity image at a plurality of spatially modulated instances to obtain a three-dimensional phase representation of the specimen at each of the plurality of distances relative to a fiducial plane; and
e. a computer code module for solving for a quantitative phase contrast tomographic image of the specimen by an inverse scattering solution, by iterative search for a point spread function entirely in a spectral domain and subject to a regularization parameter, based on a linear forward model of scattering through an imaging system.