Miniaturized multiphoton microscope with a 24Hz frame-rate

Liu, Tzu-Ming; Chan, Ming-Che; Chen, I-Hsiu; Chia, Shih‐Hsuan; Sun, Chi‐Kuang

doi:10.1364/oe.16.010501

Cited by 19 publications

(5 citation statements)

References 11 publications

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“…Compared with other techniques, such as μ-OCT or spectrally-encoded microscopy, technical challenges for the implementation of endoscopy include miniaturized high-NA (NA > 0.8) focusing objectives, fibers with low nonlinearity, good THG collection efficiency with the same fiber, miniaturized scanning mechanism, and a compact laser system. In our preliminary studies, we have built a compact Cr:forsterite laser system [24], miniaturized the scanning system by a micro-electro-mechanical system mirror [25,26], used a multimode fiber to reduce pulse broadening [26], and successfully obtained sufficient THG signals through the same fiber [26]. Therefore, we believe THG endoscopy for the examination of leukocytes in the human body can be realized in the near future.…”

Section: Discussionmentioning

confidence: 99%

Imaging granularity of leukocytes with third harmonic generation microscopy

Tsai¹,

Chen²,

Wu³

et al. 2012

Biomed. Opt. Express

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Using third harmonic generation (THG) microscopy, we demonstrate that granularity differences of leukocytes can be revealed without a label. Excited by a 1230 nm femtosecond laser, THG signals were generated at a significantly higher level in neutrophils than other mononuclear cells, whereas signals in agranular lymphocytes were one order of magnitude smaller. Interestingly, the characteristic THG features can also be observed in vivo to track the newly recruited leukocytes following lipopolysaccharide (LPS) challenge. These results suggest that label-free THG imaging may provide timely tracking of leukocyte movement without disturbing the normal cellular or physiological status.

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Section: Discussionmentioning

confidence: 99%

Imaging granularity of leukocytes with third harmonic generation microscopy

Tsai¹,

Chen²,

Wu³

et al. 2012

Biomed. Opt. Express

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“…Lissajous scanning patterns are often employed in laser scanning systems where the more common raster scanning pattern is impractical. This is often the case in miniaturized laser scanning microscopes using MEMS scanning mirrors or piezoelectric fiber scanners for beam scanning [18,20,22,[31][32][33] since, with appropriate scanning frequencies, the desired FOV can be scanned completely by Lissajous patterns. Usage of Lissajous scanning has led to rapid imaging in atomic force microscopy and multiphoton microscopy [29,31].…”

Section: Lissajous Scanningmentioning

confidence: 99%

Multiscanning mode laser scanning confocal microscopy system

Aktürk¹,

Gümüş²,

Sarioğlu³

et al. 2019

Turk J Elec Eng & Comp Sci

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In this paper, a table-top, reflective mode, laser scanning confocal microscopy system that is capable of scanning the target specimen alternately through various scanning devices and methods is proposed. We have developed a laser scanning confocal microscopy system to utilize combinations of various scanning devices and methods and to be able to characterize the optical performance of different scanners and micromirrors that are frequently used in scanning microscopy systems such as multiphoton microscopy, optical coherence tomography, or confocal microscopy. By integrating the scanner to be characterized on the same optical path with a galvanometric scan mirror, which is the conventional benchmarking scanning unit in a typical scanning microscope, we obtain two major advantages: (1) microscopy images are automatically acquired from the same location on the target specimen without having any time- consuming alignment problem and accordingly provide a high-quality optical comparison opportunity, and (2) it totally eliminates the utilization of a second scanning microscopy to benchmark the performance of the scanner-based system and considerably reduces the time spent for imaging, which is a crucial factor for a freshly excised tissue, especially under a fluorescence microscope. The system is composed of a 658 nm laser source, collimation optics, a 2D galvanometer, a 2D polymer micro-scanner, an objective lens with a numerical aperture of 0.40, a 100 μm pinhole, a PMT, a DAQ card and peripheral electronics as well as a Matlab software that simultaneously controls the system through a personal computer. Prototype of the proposed flexible LSCM system is first optically characterized using a USAF resolution target. Subsequently, we provided images of red blood and bacteria cells to demonstrate the systems’ capability for clinical diagnostics. It is reported that maximum FOV and lateral resolution of the proposed LSCM are measured to be 420 μm x 360 μm and 1 μm with galvanometer and, and 117 μm x 117 μm and 3.2 μm with the polymer scanner unit, respectively.

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“…Other applications include light barriers 11 , 3D television and mature display technologies 12 , imaging cellular network dynamics 13 , and lunar global positioning and communication systems 14 . It requires a high frame rate 15 and low spatial parameters such as angular resolution. There is no coupling between parameters in these two requirements, and the design of the Lissajous scanner is relatively simple.…”

Section: Introductionmentioning

confidence: 99%

Design rules for dense and rapid Lissajous scanning

2020

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Lissajous microscanners are very popular in compact laser-scanning applications, such as solid-state light detection and ranging (LIDAR), owing to their high-quality factor and low power consumption. In the Lissajous scanner driven by a two-axis micro-electro-mechanical system scanning mirror (MEMS-SM), the design theory is insufficient to meet the temporal and spatial resolution at the same time. In this paper, the greatest common divisor of the two-axis driving frequency is used as the temporal resolution, the concept of the fill factor (FF) is used to describe the spatial resolution of the scanner, and a general algorithm for calculating the FF is presented. Combined with the characteristics of the Lissajous trajectory, three design rules of the general Lissajous scanner are proposed, and the design theory of the Lissajous scanner enabling MEMS LIDAR is perfected. Experimental results show that the proposed design rules can effectively meet the LIDAR design requirements.

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Miniaturized multiphoton microscope with a 24Hz frame-rate

Cited by 19 publications

References 11 publications

Imaging granularity of leukocytes with third harmonic generation microscopy

Imaging granularity of leukocytes with third harmonic generation microscopy

Multiscanning mode laser scanning confocal microscopy system

Design rules for dense and rapid Lissajous scanning

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