Dual-comb Microscopy Research Articles

Coherent linking between confocal amplitude image and confocal phase image in dual-comb microscopy

We propose a method for integrating confocal amplitude and phase images obtained through dual-comb microscopy (DCM). DCM combines the benefits of confocal laser microscopy and quantitative phase microscopy, offering high axial resolution and scan-less imaging. By leveraging the coherence between confocal amplitude and phase images within the same DCM system, we accurately determine the number of phase wrapping iterations, thereby eliminating ambiguity in phase wrapping. We demonstrate this approach using samples with micrometer-range optical thickness and nanometer-scale surface roughness. The results demonstrate an expanded axial range, spanning from micrometers to millimeters, while maintaining nanometer-level axial resolution. This integrated DCM imaging technique enables the simultaneous acquisition of confocal amplitude image and absolute phase image, thus enhancing its potential for wide-axial-dynamic-range imaging across various applications.

Scan-less 3D microscopy based on spatiotemporal encoding on a single-cavity dual-comb laser.

Dual-comb microscopy enables high-speed and high-precision optical sampling by simultaneously extracting both amplitude and phase information from the interference signals with frequency division multiplexing. In this Letter, we introduce a spatiotemporal encoding approach for dual-comb microscopy that overcomes previous limitations such as mechanical scanning, low sampling efficiency, and system complexity. By employing free-space angular-chirp-enhanced delay (FACED) and a low-noise single-cavity dual-comb laser, we achieve scan-less 3D imaging with nanometer precision and a 3D distance-imaging rate of 330 Hz, restricted only by the repetition rate difference of the dual-comb laser. Specifically, the FACED unit linearly arranges the laser beam into an array. A grating subsequently disperses this array transversely into lines, facilitating ultrafast spectroscopic applications that are 1-2 orders of magnitude quicker than traditional dual-comb methods. This spatiotemporal encoding also eases the stringent conditions on various dual-comb laser parameters, such as repetition rates, coherence, and stability. Through carefully designed experiments, we demonstrate that our scan-less system can measure 3D profiles of microfabricated structures at a rate of 7 million pixels per second. Our method significantly enhances measurement speed while maintaining high precision, using a compact light source. This advancement has the potential for broad applications, including phase imaging, surface topography, distance ranging, and spectroscopy.

Computationally image-corrected dual-comb microscopy with a free-running single-cavity dual-comb fiber laser.

Dual-comb microscopy (DCM), an interesting imaging modality based on the optical-frequency-comb (OFC) mode and image pixel one-to-one correspondence, benefits from scan-less full-field imaging and simultaneous confocal amplitude and phase imaging. However, the two fully frequency-stabilized OFC sources requirement hampers DCM practicality due to the complexity and costs. Here, a bidirectional single-cavity dual-comb fiber laser (SCDCFL) is adopted as a DCM low-complexity OFC source. Although the residual timing jitter in the SCDCFL blurs the image of a static object acquired by DCM, computational image correction significantly suppresses the image blur. Nanometer-order step surface profilometry with a 14.0 nm uncertainty highlights the computationally image-corrected DCM effectiveness. We further discuss a possibility to expand the computational image correction to a dynamic object and demonstrate its preliminary experiment. The proposed method enhances the DCM generality and practicality due to low-complexity OFC source.

Full-field fluorescence lifetime dual-comb microscopy using spectral mapping and frequency multiplexing of dual-comb optical beats.

Fluorescence lifetime imaging microscopy (FLIM) is a powerful tool for quantitative fluorescence imaging because fluorescence lifetime is independent of concentration of fluorescent molecules or excitation/detection efficiency and is robust to photobleaching. However, since most FLIMs are based on point-to-point measurements, mechanical scanning of a focal spot is needed for forming an image, which hampers rapid imaging. Here, we demonstrate scan-less full-field FLIM based on a one-to-one correspondence between two-dimensional (2D) image pixels and frequency-multiplexed radio frequency (RF) signals. A vast number of dual-comb optical beats between dual optical frequency combs are effectively adopted for 2D spectral mapping and high-density frequency multiplexing in the RF region. Bimodal images of fluorescence amplitude and lifetime are obtained with high quantitativeness from amplitude and phase spectra of fluorescence RF comb modes without the need for mechanical scanning. The parallelized FLIM will be useful for rapid quantitative fluorescence imaging in life science.

Optical image amplification in dual-comb microscopy

Dual-comb microscopy (DCM), based on a combination of dual-comb spectroscopy (DCS) with two-dimensional spectral encoding (2D-SE), is a promising method for scan-less confocal laser microscopy giving an amplitude and phase image contrast with the confocality. However, signal loss in a 2D-SE optical system hampers increase in image acquisition rate due to decreased signal-to-noise ratio. In this article, we demonstrated optical image amplification in DCM with an erbium-doped fiber amplifier (EDFA). Combined use of the image-encoded DCS interferogram and the EDFA benefits from not only the batch amplification of amplitude and phase images but also significant rejection of amplified spontaneous emission (ASE) background. Effectiveness of the optical-image-amplified DCM is highlighted in the single-shot quantitative nanometer-order surface topography and the real-time movie of polystyrene beads dynamics under water convection. The proposed method will be a powerful tool for real-time observation of surface topography and fast dynamic phenomena.

Scan-Less, Kilo-Pixel, Line-Field Confocal Phase Imaging with Spectrally Encoded Dual-Comb Microscopy

Confocal laser microscopy (CLM) is a powerful tool in life science research and industrial inspection, and its image acquisition rate is boosted by scan-less imaging techniques. However, optical-intensity-based image contrast in CLM makes it difficult to visualize transparent non-fluorescent objects or reflective objects with nanometer unevenness. In this paper, we introduce an optical frequency comb (OFC) to scan-less CLM to give optical-phase-based image contrast. One-dimensional (1-D) image pixels of a sample are separately encoded onto OFC modes via 1-D spectral encoding by using OFC as an optical carrier of amplitude and phase with a vast number of discrete frequency channels. Then, line-field confocal information of amplitude and phase are decoded from mode-resolved OFC amplitude and phase spectra obtained by dual-comb spectroscopy. The proposed confocal phase imaging will further expand the applications of CLM.

Scan-less confocal phase imaging based on dual-comb microscopy

Confocal imaging and phase imaging are powerful tools in life science research and industrial inspection. To coherently link the two techniques with different depth resolutions, we introduce an optical frequency comb (OFC) to microscopy. Two-dimensional (2D) image pixels of a sample were encoded onto OFC modes via 2D spectral encoding, in which OFC acted as an optical carrier with a vast number of discrete frequency channels. Then, a scan-less full-field confocal image with a depth resolution of 62.4 um was decoded from a mode-resolved OFC amplitude spectrum obtained by dual-comb spectroscopy. Furthermore, a phase image with a depth resolution of 13.7 nm was decoded from a mode-resolved OFC phase spectrum under the above confocality. The phase wrapping ambiguity can be removed by the match between the confocal depth resolution and the phase wrapping period. The proposed hybrid microscopy approach will be a powerful tool for a variety of applications.