Method comprising determining a quantitative dispersion image of an object and digital in-line hologram microscope scanner
A method comprising determining a quantitative dispersion image of an object based on a set of quantitative phase images, each quantitative phase image of the set of quantitative phase images having been obtained with a respective different illumination light wavelength.
1 . A method comprising:
determining a quantitative dispersion image of an object based on a set of quantitative phase images, each quantitative phase image of the set of quantitative phase images having been obtained with a respective different illumination light wavelength,
wherein the set of quantitative phase images comprises three quantitative phase images, wherein each of the three phase images having been obtained with one of three different illumination light wavelengths which are ordered as, wherein λ short is the shortest wavelength of the three different wavelengths, λ middle is the middle wavelength of the three different wavelengths and Δ long is the longest wavelength of the three different wavelengths.
2 . The method of claim 1 further comprising, calculating, for each different illumination light wavelength, the respective phase image based on respective one or more phase-shifted holograms of the object.
3 . The method of claim 2 further comprising, determining a quantitative phase image of the object for each three different illumination light wavelengths by applying a Gerchberg-Saxton algorithm to the respective one or more phase-shifted holograms of the object for each three different illumination light wavelengths.
4 . The method of claim 1 further comprising, acquiring, for each different illumination light wavelength, respectively one or more phase-shifted holograms of the object at an image sensor.
5 . The method of claim 4 , wherein the respective one or more phase-shifted holograms of the object are acquired time sequentially for each of the different illumination light wavelengths and wherein the image sensor is a monochrome image sensor.
6 . The method of claim 4 , wherein the acquiring of two or more phase-shifted holograms of the object comprises shifting the distance between the image sensor and the object to realize different phase shifts.
7 . The method of claim 4 , wherein the acquiring of two or more phase-shifted holograms of the object comprises tuning a tunable phase-shifter which is placed between the object and the image sensor to realize different phase shifts.
8 . The method of claim 4 , wherein the acquiring of two or more phase-shifted holograms of the object comprises inserting different swappable elements with different refractive indices between the image sensor and the object to realize different phase shifts or comprises inserting different optical elements with different thickness between the image sensor and the object to realize different phase shifts.
9 . The method of claim 4 , wherein the acquiring of two or more phase-shifted holograms of the object comprises switching a polarizer placed on top of a birefringent optical element which are placed between the object and the image sensor to realize different phase shifts.
10 . The method of claim 1 , wherein the determining the quantitative dispersion image of the object comprises calculating, for each of different illumination light wavelengths and for each pixel of the quantitative dispersion image, an optical path difference based on a phase delay value of a respective pixel of the respective quantitative phase image.
11 . The method of claim 10 , wherein the determining a quantitative dispersion image of the object comprises calculating, for each of different illumination light wavelengths and for each pixel of the quantitative dispersion image, a refractive index based on a predetermined refractive index of a reference medium and the optical path difference of a respective pixel of the respective quantitative phase image.
12 . The method of claim 11 , wherein the determining a quantitative dispersion image of the object comprises calculating, for each pixel of the quantitative dispersion image, a quantitative dispersion value based on the respective refractive indices of the different illumination light wavelengths.
13 . The method of claim 1 , wherein the determining a quantitative dispersion image of the object comprises calculating, for each pixel of the quantitative dispersion image, a quantitative dispersion value, based on three refractive indices corresponding to the three different illumination light wavelengths
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object
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n
middle
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object
-
1
n
short
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object
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n
long
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object
wherein the first refractive index n short of three refractive indices n short , n middle , n long corresponds to shortest illumination light wavelength λshort, the second refractive index n middle of three refractive indices n short , n middle , n long corresponds to middle illumination light wavelength λ middle and the third refractive index n long of three refractive indices n short , n middle , n long corresponds to the longest illumination light wavelength Δ long .
14 . The method of claim 1 , wherein a virtual staining of the object is based on the quantitative dispersion image of the object.
15 . The method of claim 1 , wherein the three different illumination light wavelengths are blue, green and red.
16 . The method of claim 15 further comprising, calculating, for each different illumination light wavelength, a respective amplitude image based on respective one or more phase-shifted holograms of the object, and reconstructing an RGB image of the object based on the amplitude images.
17 . The method of claim 16 , wherein the virtual staining of the object is based on the quantitative dispersion image of the object and/or the RGB image of the object and/or the qualitative phase image for each of the three different illumination light wavelengths.
18 . The method of claim 1 , wherein the object is a tissue specimen.
19 . An electronic device comprising circuitry configured to:
determine a quantitative dispersion image of an object based on a set of quantitative phase images, each quantitative phase image of the set of quantitative phase images having been obtained with at least three different illumination light wavelengths which are ordered as, wherein λ short is the shortest wavelength of the three different wavelengths, λ middle is the middle wavelength of the three different wavelengths and λ long is the longest wavelength of the three different wavelengths.
20 . A digital in-line hologram microscope scanner comprising:
an image sensor configured to acquire, for each illumination light wavelength of a set of different illumination light wavelengths, respective two or more phase-shifted holograms of an object, wherein the distance between the object and the image sensor is fixed; and
circuitry configured to;
determine a set of quantitative phase images from the acquired phase-shifted holograms; and
calculate, for each of different illumination light wavelengths and for each pixel of the quantitative dispersion image, an optical path difference based on a phase delay value of a respective pixel of the respective quantitative phase image.
21 . The digital in-line hologram microscope scanner of claim 20 further comprising, a tunable phase-shifter placed between the image sensor and the object, configured to adjust the phase of emitted light, for acquiring the two or more phase-shifted holograms of the object.
22 . The digital in-line hologram microscope scanner of claim 20 further comprising, different swappable optical elements with different refractive indices placeable between the image sensor and the object, for acquiring two or more phase-shifted holograms of the object or different optical elements with different thickness between the image sensor and the object to realize different phase shifts for acquiring two or more phase-shifted holograms of the object.
23 . The digital in-line hologram microscope scanner of claim 20 further comprising, a switchable polarizer and a birefringent optical element placed between the image sensor and the object, configured to switch the polarizer for acquiring two or more phase-shifted holograms of the object.
24 . The digital in-line hologram microscope scanner of claim 20 ,
wherein the set of quantitative phase images comprises three quantitative phase images, wherein each of the three phase images having been obtained with one of three different illumination light wavelengths which are ordered as, wherein λ short is the shortest wavelength of the three different wavelengths, λ middle is the middle wavelength of the three different wavelengths and λ long is the longest wavelength of the three different wavelengths.