Integrated imaging assembly and method for using
Certain disclosed embodiments concern an integrated imaging system that combined light-sheet microscopy, which enables considerable speed and phototoxicity gains, with quantitative-phase imaging. A method for using such imaging systems also is disclosed. In an exemplary embodiment, an integrated imaging system was used for multivariate investigation of live-cells in microfluidics.
1. An integrated imaging system, comprising an integrated quantitative-phase imaging system and a light-sheet imaging system, wherein the imaging system is configured to combine spatial light interference microscopy for quantitative-phase imaging with an Airy beam light-sheet for 3D fluorescent imaging on to a standard inverted microscope.
2. The integrated imaging system according to claim 1 , comprising:
a first detection objective for quantitative-phase imaging;
a second objective for guiding light-sheet illumination, wherein the first and second objectives are configured so that illumination and detection are arranged orthogonally; and
a device for viewing, generating and/or capturing an image.
3. The integrated imaging system according to claim 2 , wherein the second objective for guiding light-sheet illumination includes a diffraction-free beam, wherein the diffraction-free beam is an Airy beam, a Bessel beam, an optical lattice or a Gaussian beam.
4. The integrated imaging system according to claim 3 wherein the Airy beam, Bessel beam, optical lattice, or Gaussian beam are generated by a spatial light modulator or a diffractive optical element.
5. The integrated imaging system according to claim 1 comprising an Airy beam generated by a spatial light modulator and configured to scan a sample in a selected direction.
6. The integrated imaging system according to claim 1 wherein transmission is encoded by a spatial light modulator to reconstruct an optical-phase image.
7. The integrated imaging system according to claim 1 further comprising a reflecting dichroic filter to direct a fluorescent signal to a first camera and transmitted white light to a spatial light modulator and a second camera.
8. The integrated imaging system according to claim 1 further comprising a microfluidic system for housing a sample, wherein the microfluidic system is manufactured using a material or materials having a refractive index or indices substantially matching the refractive index of a sample medium.
9. The integrated imaging system according to claim 8 wherein the sample medium is aqueous.
10. The integrated imaging system according to claim 8 wherein the sample comprises cells and/or tissue, and the material has a refractive index of about 1.3.
11. The integrated imaging system according to claim 8 wherein the material is a polymer selected from a MY POLYMER material, agarose, polyacrylamide, or combinations thereof.
12. The integrated imaging system according to claim 1 configured to include objectives having different levels of magnification from 1× to 100×.
13. The integrated imaging system according to claim 1 comprising a spatial light modulator to encode transmission to reconstruct an optical-phase image.
14. The integrated imaging system according to claim 2 wherein the device for viewing, generating and/or capturing an image is a camera.
15. The integrated imaging system according to claim 1 , comprising:
a first detection objective for quantitative-phase imaging;
a second objective for guiding an Airy beam generated by a first spatial light modulator to a sample enclosed in a microfluidic system, wherein the first and second objectives are arranged orthogonally;
a condenser to guide white illumination to the sample;
a second spatial light modulator to encode transmission to reconstruct an optical-phase image; and
a reflecting dichroic filter to direct a fluorescent signal to a first camera and transmitted white light to the second spatial light modulator and a second camera.
16. The integrated imaging system according to claim 2 comprising a microscope that provides the detection objective, the microscope coupled to a microscope stage configured to position a sample for scanning in 3 directions.
17. The integrated imaging system according to claim 16 wherein the microscope stage includes a piezo stage integrated with two linear stages.
18. The integrated imaging system according to claim 17 further comprising a MEMS mirror for directing an illumination beam to the sample, wherein the piezo stage, linear stages and MEMS mirror are synchronized using a controller to provide 3-dimensional imaging by scanning vertically using the piezo stage and scanning an illumination beam planarly using the MEMS mirror.
19. An imaging system, comprising:
a first detection objective to provide QPI and having different selectable levels of magnification from 1× to 100×;
a second objective having different levels of magnification from 1× to 100× for guiding an Airy beam, Bessel beam, optical lattice, or gaussian beam generated by a first spatial light modulator (SLM) for illuminating a sample housed in a microfluidic sample receiver with an illumination beam, the sample receiver comprising a material or materials having a refractive index or indices substantially matching the refractive index of a sample or sample medium, wherein the first and second objectives are configured so that illumination and detection are arranged orthogonally;
a condenser to guide white illumination to the sample;
a light conduit arranged parallel to the detection objective;
a second SLM to reconstruct an optical-phase image;
a reflecting dichroic filter to direct a fluorescent signal to a first sCMOS1 and transmitted white light to the second SLM and a second sCMOS2; and
a device for viewing, generating and/or capturing an image or images produced by the system.
20. The imaging system according to claim 19 wherein the detection objective is provided by a microscope coupled to a microscope stage configured to position a sample for scanning in 3 directions, wherein the microscope stage includes a piezo stage integrated with two linear stages, the system further comprising a MEMS mirror for guiding the illumination beam to the sample, wherein the piezo stage, linear stages and MEMS mirror are synchronized using a controller to provide 3-dimensional imaging.
21. A method, comprising:
providing an imaging system according to claim 1 ; and
using the system.
22. The method according to claim 21 , comprising using the system for imaging a biological system.
23. The method according to claim 22 , comprising imaging a cell, imaging an organelle, imaging an organ, or imaging an organism.
24. The method according to claim 23 comprising imaging a live cell sample.
25. The method according to claim 24 wherein QPI provides label-free information concerning location, size and dry-density of cells and organelles and Airy LSI provides information concerning 3D protein dynamics.
26. The method according to claim 23 comprising imaging a single cell.
27. The method according to claim 21 , comprising imaging a chemical reaction medium or constituents thereof.