Systems and methods of dual-plane digital holographic microscopy

1. An in-line digital holographic microscopy system for recording phase objects, comprising:

a light source configured to emit coherent optical waves;

a first beam splitter configured to split the coherent optical waves into object waves and reference waves;

an object area configured to accommodate the phase object, wherein the object area is configured to be illuminated by the object waves;

a second beam splitter configured to combine the object waves and the reference waves along a common optical pathway to generate interfered optical waves, wherein there is no angle between the object waves and the reference waves downstream of the second beam splitter; and

first and second imaging devices configured to record the interfered optical waves at first and second recording planes, the first recording plane positioned at a distance z from an image plane and the second recording plane positioned at a distance z+Δz from the image plane, to generate a first interferogram and a second interferogram.

2. The system of claim 1 , further comprising a computing device configured to recover phase structure of the phase object using the first interferogram and the second interferogram.

3. The system of claim 2 , wherein the computing device recovers the phase structure of the phase object by computing a first zero-mean interferogram from the first interferogram and a second zero-mean interferogram from the second interferogram and by using an operator that relates the first zero-mean interferogram and the second zero-mean interferogram.

4. The system of claim 1 , wherein the first and second imaging devices include a charge coupled device (CCD) sensor, and further wherein the light source includes a laser.

5. The system of claim 1 , wherein the second beam splitter is further configured to provide the interfered optical waves in a first set of interfered optical waves and a second set of interfered optical waves, wherein the first set of interfered optical waves is recorded using one of the first and second imaging devices at the first recording plane and the second set of interfered optical waves is recorded using one of the first and second imaging devices at the second recording plane.

6. The system of claim 1 , wherein the first and second imaging devices are configured to record the first interferogram and the second interferogram sequentially by recording the first interferogram at a first time instance at the first recording plane and subsequently recording the second interferogram at a second time instance at the second recording plane.

7. An in-line digital holographic microscopy system, comprising:

a light source configured to emit coherent optical waves;

an object area configured to accommodate a specimen, wherein the object area is configured to be illuminated by the coherent optical waves;

a first optical Fourier element configured to Fourier transform the coherent optical waves received from the object area, wherein the coherent optical waves received from the object area include directly transmitted waves and diffracted waves;

a liquid crystal phase modulator configured to introduce a phase delay between the directly transmitted waves and the diffracted waves along a common optical pathway;

a second optical Fourier element configured to receive the directly transmitted waves and the diffracted waves from the liquid crystal phase modulator and to inversely Fourier transform the directly transmitted waves and the diffracted waves, thereby providing interfered optical waves; and

first and second imaging devices configured to record the interfered optical waves at first and second recording planes, the first recording plane positioned at a distance z from an image plane and the second recording plane positioned at a distance z+Δz from the image plane, to simultaneously generate a first interferogram and a second interferogram, respectively.

8. The system of claim 7 , further comprising a computing device configured to compute specimen information using the first interferogram and the second interferogram.

9. The system of claim 8 , wherein the computing device computes the specimen information by computing a first zero-mean interferogram from the first interferogram and a second zero-mean interferogram from the second interferogram and by using an operator that relates the first zero-mean interferogram and the second zero-mean interferogram.

10. The system of claim 7 , wherein the liquid crystal phase modulator introduces the phase delay to the diffracted waves, the phase delay being substantially close to

π

2

.

11. The system of claim 7 , wherein the liquid crystal phase modulator includes a nematic liquid crystal cell that has a glass plate and a transparent electrode, wherein a small circular area at a center of the transparent electrode is removed to introduce the phase delay only to the diffracted waves.

12. The system of claim 7 , wherein the liquid crystal phase modulator introduces the phase delay to the directly transmitted waves.

13. The system of claim 7 , further comprising a beam splitter configured to split the interfered optical waves into a first set of interfered optical waves and a second set of interfered optical waves.

14. The system of claim 7 , wherein the first optical Fourier element and the second optical Fourier element include lenses, wherein the light source includes a laser, and wherein the first and second imaging devices include a charge coupled device (CCD) sensor.

15. The system of claim 7 , further comprising a dichroic mirror configured to reflect the directly transmitted waves and the diffracted waves.

16. The system of claim 15 , wherein the first and second imaging devices are further configured to record the optical waves transmitted from the dichroic mirror to generate a fluorescence image.

17. A method of analyzing phase information of a specimen, the method comprising:

generating coherent optical waves;

irradiating an object area with the coherent optical waves;

Fourier transforming the coherent optical waves received from the object area, wherein the coherent optical waves received from the object area include directly transmitted waves and diffracted waves;

introducing a phase delay between the directly transmitted waves and the diffracted waves;

upon introducing the phase delay, inversely Fourier transforming the directly transmitted waves and the diffracted waves; and

simultaneously recording the inversely Fourier transformed waves at two recording planes, the first recording plane positioned at a distance z from an image plane and the second recording plane positioned at a distance z+Δz from the image plane, to generate a first interferogram and a second interferogram.

18. The method of claim 17 , wherein the step of Fourier transforming the coherent optical waves received from the object area includes receiving the coherent optical waves from the object area using a lens.

19. The method of claim 17 , further comprising computing a phase information of the specimen using the first interferogram and the second interferogram.

20. The method of claim 19 , wherein the step of computing the phase information of the specimen using the first interferogram and the second interferogram includes computing a first zero-mean interferogram from the first interferogram and a second zero-mean interferogram from the second interferogram and using an operator that relates the first zero-mean interferogram and the second zero-mean interferogram.

21. The method of claim 17 , wherein the step of introducing the phase delay between the directly transmitted waves and the diffracted waves includes passing the directly transmitted waves through a cell with liquid crystal molecules.

22. An in-line multi-modal digital holographic microscopy system, comprising:

a light source configured to emit coherent optical waves;

an object area configured to accommodate a specimen, wherein the object area is configured to be illuminated by the coherent optical waves;

a first optical Fourier element configured to Fourier transform the coherent optical waves received from the object area, wherein the coherent optical waves received from the object area include directly transmitted waves, diffracted waves, and fluorescence waves;

a liquid crystal phase modulator configured to introduce a phase delay between the directly transmitted waves and the diffracted waves;

a second optical Fourier element configured to receive the directly transmitted waves, the diffracted waves, and the fluorescence waves, and to inversely Fourier transform the directly transmitted waves, the diffracted waves, and the fluorescence waves;

a dichroic mirror configured to reflect the directly transmitted waves and the diffracted waves, and further configured to transmit the fluorescence waves with insubstantial attenuation; and

a plurality of imaging devices configured to simultaneously record a plurality of interferograms of the directly transmitted waves and the diffracted waves at first and second recording planes, the first recording plane positioned at a distance z from an image plane and the second recording plane positioned at a distance z+Δz from the image plane, and to record the fluorescence waves at a third recording plane.

23. The system of claim 1 , wherein Δz is about 0.1 mm.