High-resolution infrared imaging of biological samples with third-order sum-frequency generation microscopy

Hanninen, Adam M.; Prince, Richard C.; Ramos, Raúl; Plikus, Maksim V.; Potma, Eric O.

doi:10.1364/boe.9.004807

Cited by 28 publications

(19 citation statements)

References 33 publications

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“…By revealing the chemical composition and thermodynamic information in addition to quantitative phase imaging, BSTP imaging promises wide applications in biology and materials science. Furthermore, by using mid-infrared excitation at the fingerprint region (800–1800 cm −1 ) where the strongest IR spectroscopy modes are located 48 , richer molecular information and stronger BSTP signals will be expected, endowing phase imaging with quantitative chemical information from vibrational spectroscopy to pave a new way for biology and materials research.…”

Section: Discussionmentioning

confidence: 99%

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: 99%

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

et al. 2019

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“…Lock-in detection arms can be refitted with the appropriate optical filters for transient absorption imaging (72). Sum-frequency generation imaging can also be readily integrated for studying ordered molecular orientation and interfacial phenomena in biological samples (73). The widefield reflectance imaging arm may be useful for integrating laser speckle, diffuse reflectance, optical coherence tomography and microscopy, or spatial frequency domain imaging with the correct illumination optics to map tissue blood flow, oxygenation, and optical properties.…”

Section: Discussionmentioning

confidence: 99%

Multi-modal Nonlinear Optical and Thermal Imaging Platform for Label-Free Characterization of Biological Tissue

Adams

Mehl

Lieser

et al. 2020

Preprint

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14The ability to characterize the combined structural, functional, and thermal properties of 15 biophysically dynamic samples is needed to address critical questions related to tissue structure, 16physiological dynamics, and disease progression. Towards this, we have developed an imaging 17 platform that enables multiple nonlinear imaging modalities to be combined with thermal 18imaging on a common sample. Here we demonstrate label-free multimodal imaging of live cells, 19excised tissues, and live rodent brain models. While potential applications of this technology are 20 wide-ranging, we expect it to be especially useful in addressing biomedical research questions 21 aimed at the biomolecular and biophysical properties of tissue and their physiology. 22

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“…Alternatively, the vibration-sensitive sum frequency generation (SFG) and thirdorder SFG processes have been harnessed to perform IR-sensitive imaging with submicrometer spatial resolution by probing the sample's nonlinear susceptibility  (2) or  (3) (22). However, these techniques have not been widely applied because of noncentrosymmetry sample requirements or the limited cross section of the nonlinear processes (23,24).…”

Section: Introductionmentioning

confidence: 99%

Bond-selective imaging by optically sensing the mid-infrared photothermal effect

2021

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Mid-infrared (IR) spectroscopic imaging using inherent vibrational contrast has been broadly used as a powerful analytical tool for sample identification and characterization. However, the low spatial resolution and large water absorption associated with the long IR wavelengths hinder its applications to study subcellular features in living systems. Recently developed mid-infrared photothermal (MIP) microscopy overcomes these limitations by probing the IR absorption–induced photothermal effect using a visible light. MIP microscopy yields submicrometer spatial resolution with high spectral fidelity and reduced water background. In this review, we categorize different photothermal contrast mechanisms and discuss instrumentations for scanning and widefield MIP microscope configurations. We highlight a broad range of applications from life science to materials. We further provide future perspective and potential venues in MIP microscopy field.

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High-resolution infrared imaging of biological samples with third-order sum-frequency generation microscopy

Cited by 28 publications

References 33 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

Multi-modal Nonlinear Optical and Thermal Imaging Platform for Label-Free Characterization of Biological Tissue

Bond-selective imaging by optically sensing the mid-infrared photothermal effect

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