Molecular contrast on phase-contrast microscope

Toda, Keiichiro; Tamamitsu, Miu; Nagashima, Yu; Horisaki, Ryoichi; Ideguchi, Takuro

doi:10.1038/s41598-019-46383-6

Cited by 39 publications

(54 citation statements)

References 31 publications

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“…We note that two independent studies on the same topic were recently published 33,34 during the peer review of the present work. Our BSTP work was deposited on arXiv on Nov. 27, 2018 (arXiv:1811.11140), and the aforementioned publications were deposited on Dec. 11, 2018 (arXiv:1812.04576, later published in Scientific Reports), and Mar.…”

Section: Discussionmentioning

confidence: 92%

Bond-selective transient phase imaging via sensing of the infrared photothermal effect

et al. 2019

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Phase-contrast microscopy converts the phase shift of light passing through a transparent specimen, e.g., a biological cell, into brightness variations in an image. This ability to observe structures without destructive fixation or staining has been widely utilized for applications in materials and life sciences. Despite these advantages, phase-contrast microscopy lacks the ability to reveal molecular information. To address this gap, we developed a bond-selective transient phase (BSTP) imaging technique that excites molecular vibrations by infrared light, resulting in a transient change in phase shift that can be detected by a diffraction phase microscope. By developing a time-gated pump–probe camera system, we demonstrate BSTP imaging of live cells at a 50 Hz frame rate with high spectral fidelity, sub-microsecond temporal resolution, and sub-micron spatial resolution. Our approach paves a new way for spectroscopic imaging investigation in biology and materials science.

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Section: Discussionmentioning

confidence: 92%

Bond-selective transient phase imaging via sensing of the infrared photothermal effect

et al. 2019

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“…In this work, we additionally harness the capability of QPI to computationally synthesize the 3D spatial-frequency aperture of the imaging system 2,22 . This enables us to further achieve (3) the higher lateral spatial resolution by the expanded bandwidth of the synthetic aperture that surpasses the numerical aperture (NA) of a single objective lens which poses the diffraction limit in other farfield MVI techniques [6][7][8][9][10][11][12][13][14][15][16][17][18]20 and (4) decoupling the undesired MIR absorption effect of the surrounding aqueous medium, which is the well-known problem of MIR microscopy in general, by the depth-resolving power offered by the axial bandwidth of the 3D synthetic aperture.…”

Section: Mv-qpi the Concept Of Mv-qpi Is Illustrated Inmentioning

confidence: 99%

Label-free biochemical quantitative phase imaging with mid-infrared photothermal effect

Tamamitsu¹,

Toda²,

Shimada³

et al. 2020

Optica

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Label-free optical imaging is valuable in biology and medicine with its non-destructive property and reduced optical and chemical damages. Quantitative phase (QPI) and molecular vibrational imaging (MVI) are the two most successful label-free methods, providing morphology and biochemistry, respectively, that have pioneered numerous applications along their independent technological maturity over the past few decades. However, the distinct label-free contrasts are inherently complementary and difficult to integrate due to the use of different light-matter interactions. Here, we present a unified imaging scheme that realizes simultaneous and in-situ acquisition of MV-fingerprint contrasts of single cells in the framework of QPI utilizing the mid-infrared photothermal effect. The fully label-free and robust integration of subcellular morphology and biochemistry would have important implications, especially for studying complex and fragile biological phenomena such as drug delivery, cellular diseases and stem cell development, where long-time observation of unperturbed cells are needed under low phototoxicity.

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“…To illustrate the advantage of ADRIFT-QPI, we apply this technique to MIR photothermal QPI [10][11][12][13] . MIR photothermal QPI is a recently developed molecular vibrational imaging method, where the refractive index change of the sample caused by the absorption of MIR pump light is detected through the OPD change of visible Figure 4a shows the pump-OFF-state OPD images obtained by conventional DH and ADRIFT-DH.…”

Section: Dynamic-range-expanded Mir Photothermal Qpi Of Microbeadsmentioning

confidence: 99%

“…Phase imaging [1][2][3][4][5][6][7][8][9][10][11][12][13][14][15][16][17][18][19] provides morphological phasecontrast of transparent samples and is widely used in various fields, especially in biological science, because morphological features of micrometre-scale specimens provide valuable information on complex biological phenomena. Quantitative phase imaging (QPI) [1][2][3][4][5][6] is the most powerful method for studying cellular morphology among various phase imaging methods, such as phase-contrast 7 and differential-interference-contrast 8 imaging, because it is able to accurately measure the optical phase delay (OPD) caused by a sample.…”

Section: Introductionmentioning

confidence: 99%

“…Since the minimum detectable OPD of conventional QPI (~10 mrad) is already small enough to clearly observe the surface roughness of a glass slide that limits the detectable sample-induced OPD, sensitivity improvement of QPI, especially of the temporal OPD sensitivity, has not been intensively explored. However, recent developments in pump-probe-type perturbative QPI techniques, such as mid-infrared (MIR) photothermal QPI [10][11][12][13] , have shown that temporal differential analysis of consecutively measured images can cancel out the substrate background and reveal small OPD changes. Thus, sensitivity improvement of QPI is highly demanded in this context.…”

Section: Introductionmentioning

confidence: 99%

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Adaptive dynamic range shift (ADRIFT) quantitative phase imaging

2021

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Quantitative phase imaging (QPI) with its high-contrast images of optical phase delay (OPD) maps is often used for label-free single-cell analysis. Contrary to other imaging methods, sensitivity improvement has not been intensively explored because conventional QPI is sensitive enough to observe the surface roughness of a substrate that restricts the minimum measurable OPD. However, emerging QPI techniques that utilize, for example, differential image analysis of consecutive temporal frames, such as mid-infrared photothermal QPI, mitigate the minimum OPD limit by decoupling the static OPD contribution and allow measurement of much smaller OPDs. Here, we propose and demonstrate supersensitive QPI with an expanded dynamic range. It is enabled by adaptive dynamic range shift through a combination of wavefront shaping and dark-field QPI techniques. As a proof-of-concept demonstration, we show dynamic range expansion (sensitivity improvement) of QPI by a factor of 6.6 and its utility in improving the sensitivity of mid-infrared photothermal QPI. This technique can also be applied for wide-field scattering imaging of dynamically changing nanoscale objects inside and outside a biological cell without losing global cellular morphological image information.

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Molecular contrast on phase-contrast microscope

Cited by 39 publications

References 31 publications

Bond-selective transient phase imaging via sensing of the infrared photothermal effect

Bond-selective transient phase imaging via sensing of the infrared photothermal effect

Label-free biochemical quantitative phase imaging with mid-infrared photothermal effect

Adaptive dynamic range shift (ADRIFT) quantitative phase imaging

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