Label-free biochemical refractive-index tomography of single cells with mid-infrared photothermal effect

2020 ◽  
Author(s):  
Miu Tamamitsu ◽  
Keiichiro Toda ◽  
Hiroyuki Shimada ◽  
Yu Nagashima ◽  
Ryoichi Horisaki ◽  
...  
Keyword(s):  
Refractive Index ◽  
Single Cells ◽  
Photothermal Effect ◽  
Label Free ◽  
Mid Infrared

scholarly journals Depth-resolved mid-infrared photothermal imaging of living cells and organisms with submicrometer spatial resolution

2016 ◽  
Vol 2 (9) ◽  
pp. e1600521 ◽  
Author(s):  
Delong Zhang ◽  
Chen Li ◽  
Chi Zhang ◽  
Mikhail N. Slipchenko ◽  
Gregory Eakins ◽  
...  
Keyword(s):  
Spatial Resolution ◽  
Single Cells ◽  
Chemical Imaging ◽  
Detection Sensitivity ◽  
Living Systems ◽  
Live Cells ◽  
Photothermal Effect ◽  
Label Free ◽  
Infrared Microscopy ◽  
Mid Infrared

Chemical contrast has long been sought for label-free visualization of biomolecules and materials in complex living systems. Although infrared spectroscopic imaging has come a long way in this direction, it is thus far only applicable to dried tissues because of the strong infrared absorption by water. It also suffers from low spatial resolution due to long wavelengths and lacks optical sectioning capabilities. We overcome these limitations through sensing vibrational absorption–induced photothermal effect by a visible laser beam. Our mid-infrared photothermal (MIP) approach reached 10 μM detection sensitivity and submicrometer lateral spatial resolution. This performance has exceeded the diffraction limit of infrared microscopy and allowed label-free three-dimensional chemical imaging of live cells and organisms. Distributions of endogenous lipid and exogenous drug inside single cells were visualized. We further demonstrated in vivo MIP imaging of lipids and proteins inCaenorhabditis elegans. The reported MIP imaging technology promises broad applications from monitoring metabolic activities to high-resolution mapping of drug molecules in living systems, which are beyond the reach of current infrared microscopy.

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

Optica ◽  
2020 ◽  
Vol 7 (4) ◽  
pp. 359 ◽  
Author(s):  
Miu Tamamitsu ◽  
Keiichiro Toda ◽  
Hiroyuki Shimada ◽  
Takaaki Honda ◽  
Masaharu Takarada ◽  
...  
Keyword(s):  
Photothermal Effect ◽  
Phase Imaging ◽  
Label Free ◽  
Quantitative Phase Imaging ◽  
Mid Infrared ◽  
Quantitative Phase

scholarly journals 3D phonon microscopy with sub-micron axial-resolution

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Richard J. Smith ◽  
Fernando Pérez-Cota ◽  
Leonel Marques ◽  
Matt Clark
Keyword(s):  
Signal To Noise Ratio ◽  
Single Cells ◽  
Optical Resolution ◽  
Acquisition Time ◽  
Label Free ◽  
Axial Resolution ◽  
Force Microscopy ◽  
Atomic Force ◽  
Accuracy And Precision ◽  
Good Agreement

AbstractBrillouin light scattering (BLS) is an emerging method for cell imaging and characterisation. It allows elasticity-related contrast, optical resolution and label-free operation. Phonon microscopy detects BLS from laser generated coherent phonon fields to offer an attractive route for imaging since, at GHz frequencies, the phonon wavelength is sub-optical. Using phonon fields to image single cells is challenging as the signal to noise ratio and acquisition time are often poor. However, recent advances in the instrumentation have enabled imaging of fixed and living cells. This work presents the first experimental characterisation of phonon-based axial resolution provided by the response to a sharp edge. The obtained axial resolution is up to 10 times higher than that of the optical system used to take the measurements. Validation of the results are obtained with various polymer objects, which are in good agreement with those obtained using atomic force microscopy. Edge localisation, and hence profilometry, of a phantom boundary is measured with accuracy and precision of approximately 60 nm and 100 nm respectively. Finally, 3D imaging of fixed cells in culture medium is demonstrated.

scholarly journals Critical angle reflection imaging for quantification of molecular interactions on glass surface

2021 ◽  
Vol 12 (1) ◽  
Author(s):  
Guangzhong Ma ◽  
Runli Liang ◽  
Zijian Wan ◽  
Shaopeng Wang
Keyword(s):  
Refractive Index ◽  
Small Molecule ◽  
Molecular Interactions ◽  
Glass Surface ◽  
Critical Angle ◽  
Dynamic Range ◽  
Refractive Index Change ◽  
Label Free ◽  
Index Change ◽  
Sensing Range

AbstractQuantification of molecular interactions on a surface is typically achieved via label-free techniques such as surface plasmon resonance (SPR). The sensitivity of SPR originates from the characteristic that the SPR angle is sensitive to the surface refractive index change. Analogously, in another interfacial optical phenomenon, total internal reflection, the critical angle is also refractive index dependent. Therefore, surface refractive index change can also be quantified by measuring the reflectivity near the critical angle. Based on this concept, we develop a method called critical angle reflection (CAR) imaging to quantify molecular interactions on glass surface. CAR imaging can be performed on SPR imaging setups. Through a side-by-side comparison, we show that CAR is capable of most molecular interaction measurements that SPR performs, including proteins, nucleic acids and cell-based detections. In addition, we show that CAR can detect small molecule bindings and intracellular signals beyond SPR sensing range. CAR exhibits several distinct characteristics, including tunable sensitivity and dynamic range, deeper vertical sensing range, fluorescence compatibility, broader wavelength and polarization of light selection, and glass surface chemistry. We anticipate CAR can expand SPR′s capability in small molecule detection, whole cell-based detection, simultaneous fluorescence imaging, and broader conjugation chemistry.

scholarly journals Label-Free and Quantitative Dry Mass Monitoring for Single Cells during In Situ Culture

Cells ◽  
2021 ◽  
Vol 10 (7) ◽  
pp. 1635
Author(s):  
Ya Su ◽  
Rongxin Fu ◽  
Wenli Du ◽  
Han Yang ◽  
Li Ma ◽  
...  
Keyword(s):  
Cell Growth ◽  
Single Cell ◽  
Hela Cells ◽  
Quantitative Measurement ◽  
Single Cells ◽  
Dry Mass ◽  
Quantitative Detection ◽  
Label Free ◽  
Mass Sensitivity

Quantitative measurement of single cells can provide in-depth information about cell morphology and metabolism. However, current live-cell imaging techniques have a lack of quantitative detection ability. Herein, we proposed a label-free and quantitative multichannel wide-field interferometric imaging (MWII) technique with femtogram dry mass sensitivity to monitor single-cell metabolism long-term in situ culture. We demonstrated that MWII could reveal the intrinsic status of cells despite fluctuating culture conditions with 3.48 nm optical path difference sensitivity, 0.97 fg dry mass sensitivity and 2.4% average maximum relative change (maximum change/average) in dry mass. Utilizing the MWII system, different intrinsic cell growth characteristics of dry mass between HeLa cells and Human Cervical Epithelial Cells (HCerEpiC) were studied. The dry mass of HeLa cells consistently increased before the M phase, whereas that of HCerEpiC increased and then decreased. The maximum growth rate of HeLa cells was 11.7% higher than that of HCerEpiC. Furthermore, HeLa cells were treated with Gemcitabine to reveal the relationship between single-cell heterogeneity and chemotherapeutic efficacy. The results show that cells with higher nuclear dry mass and nuclear density standard deviations were more likely to survive the chemotherapy. In conclusion, MWII was presented as a technique for single-cell dry mass quantitative measurement, which had significant potential applications for cell growth dynamics research, cell subtype analysis, cell health characterization, medication guidance and adjuvant drug development.

scholarly journals Cellular lensing and near infrared fluorescent nanosensor arrays to enable chemical efflux cytometry

2021 ◽  
Vol 12 (1) ◽  
Author(s):  
Soo-Yeon Cho ◽  
Xun Gong ◽  
Volodymyr B. Koman ◽  
Matthias Kuehne ◽  
Sun Jin Moon ◽  
...  
Keyword(s):  
Near Infrared ◽  
Single Cells ◽  
Microfluidic Channel ◽  
Human Monocyte ◽  
Cellular Heterogeneity ◽  
Label Free ◽  
Nanotube Array ◽  
Chemical Cytometry ◽  
Rich Information ◽  
Non Destructive

AbstractNanosensors have proven to be powerful tools to monitor single cells, achieving spatiotemporal precision even at molecular level. However, there has not been way of extending this approach to statistically relevant numbers of living cells. Herein, we design and fabricate nanosensor array in microfluidics that addresses this limitation, creating a Nanosensor Chemical Cytometry (NCC). nIR fluorescent carbon nanotube array is integrated along microfluidic channel through which flowing cells is guided. We can utilize the flowing cell itself as highly informative Gaussian lenses projecting nIR profiles and extract rich information. This unique biophotonic waveguide allows for quantified cross-correlation of biomolecular information with various physical properties and creates label-free chemical cytometer for cellular heterogeneity measurement. As an example, the NCC can profile the immune heterogeneities of human monocyte populations at attomolar sensitivity in completely non-destructive and real-time manner with rate of ~600 cells/hr, highest range demonstrated to date for state-of-the-art chemical cytometry.

scholarly journals Perfect Absorption and Refractive-Index Sensing by Metasurfaces Composed of Cross-Shaped Hole Arrays in Metal Substrate

Nanomaterials ◽  
2020 ◽  
Vol 11 (1) ◽  
pp. 63
Author(s):  
Zhendong Yan ◽  
Chaojun Tang ◽  
Guohua Wu ◽  
Yumei Tang ◽  
Ping Gu ◽  
...  
Keyword(s):  
Refractive Index ◽  
High Performance ◽  
Narrow Band ◽  
Label Free ◽  
Hybrid Mode ◽  
Perfect Absorption ◽  
Refractive Index Sensing ◽  
Sensing Applications ◽  
Silver Substrate ◽  
Hole Arrays

Achieving perfect electromagnetic wave absorption with a sub-nanometer bandwidth is challenging, which, however, is desired for high-performance refractive-index sensing. In this work, we theoretically study metasurfaces for sensing applications based on an ultra-narrow band perfect absorption in the infrared region, whose full width at half maximum (FWHM) is only 1.74 nm. The studied metasurfaces are composed of a periodic array of cross-shaped holes in a silver substrate. The ultra-narrow band perfect absorption is related to a hybrid mode, whose physical mechanism is revealed by using a coupling model of two oscillators. The hybrid mode results from the strong coupling between the magnetic resonances in individual cross-shaped holes and the surface plasmon polaritons on the top surface of the silver substrate. Two conventional parameters, sensitivity (S) and figure of merit (FOM), are used to estimate the sensing performance, which are 1317 nm/RIU and 756, respectively. Such high-performance parameters suggest great potential for the application of label-free biosensing.

Label-free and noninvasive optical detection of the distribution of nanometer-size mitochondria in single cells

2011 ◽  
Vol 16 (6) ◽  
pp. 067003 ◽  
Author(s):  
Xuantao Su ◽  
Yuanyuan Qiu ◽  
Leah Marquez-Curtis ◽  
Manisha Gupta ◽  
Clarence E. Capjack ◽  
...  
Keyword(s):  
Single Cells ◽  
Optical Detection ◽  
Label Free ◽  
Nanometer Size

scholarly journals Label-Free Digital Holo-tomographic Microscopy Reveals Virus-Induced Cytopathic Effects in Live Cells

mSphere ◽  
2018 ◽  
Vol 3 (6) ◽  
Author(s):  
Artur Yakimovich ◽  
Robert Witte ◽  
Vardan Andriasyan ◽  
Fanny Georgi ◽  
Urs F. Greber
Keyword(s):  
Refractive Index ◽  
Vaccinia Virus ◽  
Virus Type ◽  
Live Cells ◽  
Refractive Index Gradient ◽  
Label Free ◽  
Virus Infections ◽  
Cytopathic Effects ◽  
Infected Cells ◽  
Index Gradient

ABSTRACTCytopathic effects (CPEs) are a hallmark of infections. CPEs are difficult to observe due to phototoxicity from classical light microscopy. We report distinct patterns of virus infections in live cells using digital holo-tomographic microscopy (DHTM). DHTM is label-free and records the phase shift of low-energy light passing through the specimen on a transparent surface with minimal perturbation. DHTM measures the refractive index (RI) and computes the refractive index gradient (RIG), unveiling optical heterogeneity in cells. We find that vaccinia virus (VACV), herpes simplex virus (HSV), and rhinovirus (RV) infections progressively and distinctly increased RIG. VACV infection, but not HSV and RV infections, induced oscillations of cell volume, while all three viruses altered cytoplasmic membrane dynamics and induced apoptotic features akin to those caused by the chemical compound staurosporine. In sum, we introduce DHTM for quantitative label-free microscopy in infection research and uncover virus type-specific changes and CPE in living cells with minimal interference.IMPORTANCEThis study introduces label-free digital holo-tomographic microscopy (DHTM) and refractive index gradient (RIG) measurements of live, virus-infected cells. We use DHTM to describe virus type-specific cytopathic effects, including cyclic volume changes of vaccinia virus infections, and cytoplasmic condensations in herpesvirus and rhinovirus infections, distinct from apoptotic cells. This work shows for the first time that DHTM is suitable to observe virus-infected cells and distinguishes virus type-specific signatures under noninvasive conditions. It provides a basis for future studies, where correlative fluorescence microscopy of cell and virus structures annotate distinct RIG values derived from DHTM.

Rational synthesis of hollow cubic CuS@Spiky Au core–shell nanoparticles for enhanced photothermal and SERS effects

2018 ◽  
Vol 54 (95) ◽  
pp. 13399-13402 ◽  
Author(s):  
Qian Lv ◽  
Meng-Yue Gao ◽  
Zi-He Cheng ◽  
Qiao Chen ◽  
Ai-Guo Shen ◽  
...  
Keyword(s):  
Cell Apoptosis ◽  
Photothermal Effect ◽  
Core Shell ◽  
Label Free ◽  
Shell Nanoparticles ◽  
Intracellular Imaging ◽  
Core Shell Nanoparticles

The unique “tip spots” can simultaneously enhance SERS and the photothermal effect, facilitating label-free SERS intracellular imaging during cell apoptosis.

Sign in / Sign up

Export Citation Format

Share Document