Abstract
We introduce a wide field hyperspectral microscope using the Fourier-transform approach. The interferometer is based on the translating-wedge-based identical pulses encoding system, a common-path birefringent interferometer which combines compactness, intrinsic interferometric delay precision, long-term stability, and insensitivity to vibrations. We describe two different implementations of our system, which maximize fringe visibility and phase invariance over the field of view, respectively. We also demonstrate that our system can be installed as an add-on in a commercial microscope. We show high-quality hyperspectral fluorescence microscopy from stained cells and powders of inorganic pigments in the spectral range from 400 to 1100 nm, proving that our device is suited to biology and materials science. We also introduce an acquisition method that synthesizes a tunable spectral filter, providing band-passed images with the measurement of only two maps.
Highlights
In the last few decades, a huge advancement has occurred in optical microscopy, with revolutionary improvements in resolution and capability to reconstruct the three-dimensional features of samples, from nanoscale to millimeter size.[1,2] At the same time, a disruptive transformation has occurred in biological samples that optical microscopy is demanded to analyze
We demonstrate that our system can be installed as an add-on in a commercial microscope
We have demonstrated that the Translating-Wedge-based Identical pulses eNcoding System (TWINS) interferometer is well suited to design a very compact hyperspectral microscope featuring: (i) wide spectral coverage, (ii) user adjustable spectral resolution, (iii) spatial resolution basically unaffected compared to a standard microscope, (iv) compactness, and (v) high spectral sensitivity thanks to the large contrast of the interferometer
Summary
In the last few decades, a huge advancement has occurred in optical microscopy, with revolutionary improvements in resolution and capability to reconstruct the three-dimensional features of samples, from nanoscale to millimeter size.[1,2] At the same time, a disruptive transformation has occurred in biological samples that optical microscopy is demanded to analyze Encoded probes, such as green fluorescent protein, have been generated to target proteins, intracellular ions, metabolites, messengers (H+, Ca2+, Cl−, etc.), in cells’ and organisms’ compartments.[3] molecular biologists have learned how to combine probes emitting at different wavelengths, either genetically expressed or based on immunohistochemistry, in the same specimen in order to differentiate biological structures and reveal functional interactions. Our results show that the TWINS applied to FT microscopy provides a high degree of coherence at each pixel of the image, enabling a wide-field HSM with high spectral accuracy
Sign-in/Register to view highlights & summary Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callDisclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
