Simple high-speed confocal line-scanning microscope

Im, Kang Bin; Han, Sumin; Park, Hwajoon; Kim, Dong–Sun; Kim, Beop Min

doi:10.1364/opex.13.005151

Cited by 71 publications

(52 citation statements)

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“…Further, to significantly enhance the speed of data acquisition (a modification essential for capturing CTCs in rapidly flowing blood vessels), customary two-dimensional scanning was changed to one-dimensional line scanning along a transept orthogonal to the flow of blood in the monitored vessel. This change improved the rate of data acquisition 250-fold (21).…”

Section: Evaluation and Optimization Of The Methods For In Vivo Imagimentioning

confidence: 99%

In vivo quantitation of rare circulating tumor cells by multiphoton intravital flow cytometry

He¹,

Wang

Hartmann

et al. 2007

Proc. Natl. Acad. Sci. U.S.A.

270

276

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Quantitation of circulating tumor cells (CTCs) constitutes an emerging tool for the diagnosis and staging of cancer, assessment of response to therapy, and evaluation of residual disease after surgery. Unfortunately, no existing technology has the sensitivity to measure the low numbers of tumor cells (<1 CTC per ml of whole blood) that characterize minimal levels of disease. We present a method, intravital flow cytometry, that noninvasively counts rare CTCs in vivo as they flow through the peripheral vasculature. The method involves i.v. injection of a tumor-specific fluorescent ligand followed by multiphoton fluorescence imaging of superficial blood vessels to quantitate the flowing CTCs. Studies in mice with metastatic tumors demonstrate that CTCs can be quantitated weeks before metastatic disease is detected by other means. Analysis of whole blood samples from cancer patients further establishes that human CTCs can be selectively labeled and quantitated when present at Ϸ2 CTCs per ml, opening opportunities for earlier assessment of metastatic disease.folate conjugates ͉ in vivo imaging ͉ multiphoton microscopy ͉ metastasis ͉ cancer diagnosis

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Section: Evaluation and Optimization Of The Methods For In Vivo Imagimentioning

confidence: 99%

In vivo quantitation of rare circulating tumor cells by multiphoton intravital flow cytometry

He¹,

Wang

Hartmann

et al. 2007

Proc. Natl. Acad. Sci. U.S.A.

270

276

View full text Add to dashboard Cite

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“…Using a 20 kHz line scan rate, we achieved a frame rate of 20 Hz for 512x512 pixels en-face confocal images, which is fast enough for in-vivo applications appropriate for LCM [12][13][14][15][16]. The system can be engineered to achieve higher speed if necessary.…”

Section: Discussionmentioning

confidence: 99%

“…It is promising to find more applications if the speed can be improved to tens frames per second. Instead of working with PCM using the above-mentioned phase-sensitive techniques, there has been significant interest in line-scanning confocal microscopy (LCM) because it has a simpler optical system and achieves faster data acquisition while maintaining comparable confocal imaging compared to PCM [12][13][14][15][16]. Previous work has demonstrated a quantitative phasecontrast confocal imaging technique by combining DH with a traditional line-scanning confocal microscope [17,18].…”

Section: Introductionmentioning

confidence: 99%

High-speed line-field confocal holographic microscope for quantitative phase imaging

et al. 2016

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Abstract:We present a high-speed and phase-sensitive reflectance linescanning confocal holographic microscope (LCHM). We achieved rapid confocal imaging using a fast line-scan CCD camera and quantitative phase imaging using off-axis digital holography (DH) on a 1D, line-by-line basis in our prototype experiment. Using a 20 kHz line scan rate, we achieved a frame rate of 20 Hz for 512x512 pixels en-face confocal images. We realized coherent holographic detection two different ways. We first present a LCHM using off-axis configuration. By using a microscope objective of a NA 0.65, we achieved axial and lateral resolution of ~3.5 micrometers and ~0.8 micrometers, respectively. We demonstrated surface profile measurement of a phase target at nanometer precision and the digital refocusing of a defocused confocal en-face image. Ultrahigh temporal resolution M mode is demonstrated by measuring the vibration of a PZTactuated mirror driven by a sine wave at 1 kHz. We then report our experimental work on a LCHM using an in-line configuration. In this inline LCHM, the coherent detection is enabled by moving the reference arm at a constant speed, thereby introducing a Doppler frequency shift that leads to spatial interference fringes along the scanning direction. Lastly, we present a unified formulation that treats off-axis and in-line LCHM in a unified joint spatiotemporal modulation framework and provide a connection between LCHM and the traditional off-axis DH. The presented high-speed LCHM may find applications in optical metrology and biomedical imaging.

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“…However, the speed of the image acquisition is limited by the point-scanning configuration. To speed up image acquisition and simplify the optical system, line-scanning confocal systems have been proposed and tested in industrial inspection, imaging of human tissues, and ophthalmology [6][7][8][9]. Equipped with adaptive optics, the line-scanning confocal ophthalmoscope is able to image the human retina at the cellular level [10].…”

Section: Introductionmentioning

confidence: 99%

Quantitative phase-contrast confocal microscope

Liu¹,

Marchesini²,

Kim³

2014

Opt. Express

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We present a quantitative phase-contrast confocal microscope (QPCCM) by combining a line-scanning confocal system with digital holography (DH). This combination can merge the merits of these two different imaging modalities. High-contrast intensity images with low coherent noise, and the optical sectioning capability are made available due to the confocality. Phase profiles of the samples become accessible thanks to DH. QPCCM is able to quantitatively measure the phase variations of optical sections of the opaque samples and has the potential to take highquality intensity and phase images of non-opaque samples such as many biological samples. Because each line scan is recorded by a hologram that may contain the optical aberrations of the system, it opens avenues for a variety of numerical aberration compensation methods and development of full digital adaptive optics confocal system to emulate current hardwarebased adaptive optics system for biomedical imaging, especially ophthalmic imaging. Preliminary experiments with a microscope objective of NA 0.65 and 40 × on opaque samples are presented to demonstrate this idea. The measured lateral and axial resolutions of the intensity images from the current system are ~0.64μm and ~2.70μm respectively. The noise level of the phase profile by QPCCM is ~2.4nm which is better than the result by DH.

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Simple high-speed confocal line-scanning microscope

Cited by 71 publications

References 0 publications

In vivo quantitation of rare circulating tumor cells by multiphoton intravital flow cytometry

In vivo quantitation of rare circulating tumor cells by multiphoton intravital flow cytometry

High-speed line-field confocal holographic microscope for quantitative phase imaging

Quantitative phase-contrast confocal microscope

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