Intravital imaging by simultaneous label-free autofluorescence-multiharmonic microscopy

You, Sixian; Tu, Haohua; Chaney, Eric J.; Sun, Yi; Zhao, Youbo; Bower, Andrew J.; Liu, Yuan Zhi; Marjanović, Marina; Sinha, Saurabh; Pu, Yang; Boppart, Stephen A.

doi:10.1038/s41467-018-04470-8

Cited by 222 publications

(223 citation statements)

References 59 publications

Supporting

Mentioning

215

Contrasting

Order By: Relevance

“…(iii‐iv) the deeper z‐sections of the area show a mature blood vessel containing red blood cells (red arrows). Adapted and reprinted with permission from You et al…”

Section: Translational Potential Of Iv‐mpm For Cancermentioning

confidence: 99%

“…Due to limited applications of such high‐price light sources outside of THG in microscopy, THG is not as extensively used as SHG to date (this is likely to change with the recent development of fiber lasers). In IV‐MPM of cancer, THG so far has aided in the imaging of the blood vessel walls and blood cells and provided outlines of adipocytes and extracellular vesicles …”

Section: Translational Potential Of Iv‐mpm For Cancermentioning

confidence: 99%

“…For example, the combination of 2P and 3P excitation requires sequential imaging because of the different wavelengths required, resulting in longer acquisition times and increased photodamage. To overcome such challenges, You et al designed a novel imaging platform called simultaneous label‐free autofluorescence multiharmonic (SLAM) microscopy . This methodology allows simultaneous imaging of NADH and FAD via 2P and 3P excited autofluorescence, respectively, as well as imaging of noncentrosymmetric structures via SHG signal and interfacial features with THG signal using a single excitation source.…”

Section: Translational Potential Of Iv‐mpm For Cancermentioning

confidence: 99%

“…(iii-iv) the deeper z-sections of the area show a mature blood vessel containing red blood cells (red arrows). Adapted and reprinted with permission from You et al 142 to exhibit nonmitotic effects and reduced efficacy compared with effects measured in vitro. 38 Overall, measuring responses to chemotherapeutic agents by IV-MPM may help explain varied and limited responses in clinical trials and improve current cancer treatment strategies.…”

Section: Monitoring Chemotherapy Distribution Effects and Resistamentioning

confidence: 99%

“…In IV-MPM of cancer, THG so far has aided in the imaging of the blood vessel walls and blood cells 161 and provided outlines of adipocytes 162 and extracellular vesicles. 142 Autofluorescence allows imaging of metabolites and structural tissue components. Metabolic cofactors nicotinamide adenine dinucleotide (NADH) and flavin adenine dinucleotide (FAD) are of particular interest, as they emit the strongest autofluorescence signals.…”

Section: Label-free Iv-mpmmentioning

confidence: 99%

See 4 more Smart Citations

Frontiers in intravital multiphoton microscopy of cancer

2019

View full text Add to dashboard Cite

Background Cancer is a highly complex disease, which involves the cooperation of tumor cells with multiple types of host cells and the extracellular matrix. Cancer studies that rely solely on static measurements of individual cell types are insufficient to dissect this complexity. In the last two decades, intravital microscopy has established itself as a powerful technique that can significantly improve our understanding of cancer by revealing the dynamic interactions governing cancer initiation, progression, and treatment effects in living animals. This review focuses on intravital multiphoton microscopy (IV‐MPM) applications in mouse models of cancer. Recent findings IV‐MPM studies have already enabled a deeper understanding of the complex events occurring in cancer at the molecular, cellular, and tissue levels. Multiple cell types present in different tissues influence cancer cell behavior via activation of distinct signaling pathways. As a result, the boundaries in the field of IV‐MPM are continuously being pushed to provide an integrated comprehension of cancer. We propose that optics, informatics, and cancer (cell) biology are coevolving as a new field. We have identified four emerging themes in this new field. First, new microscopy systems and image processing algorithms are enabling the simultaneous identification of multiple interactions between the tumor cells and the components of the tumor microenvironment. Second, techniques from molecular biology are being exploited to visualize subcellular structures and protein activities within individual cells of interest and relate those to phenotypic decisions, opening the door for “in vivo cell biology”. Third, combining IV‐MPM with additional imaging modalities or omics studies holds promise for linking the cell phenotype to its genotype, metabolic state, or tissue location. Finally, the clinical use of IV‐MPM for analyzing efficacy of anticancer treatments is steadily growing, suggesting a future role of IV‐MPM for personalized medicine. Conclusion IV‐MPM has revolutionized visualization of tumor‐microenvironment interactions in real time. Moving forward, incorporation of novel optics, automated image processing, and omics technologies in the study of cancer biology, will not only advance our understanding of the underlying complexities but will also leverage the unique aspects of IV‐MPM for clinical use.

show abstract

“…(iii‐iv) the deeper z‐sections of the area show a mature blood vessel containing red blood cells (red arrows). Adapted and reprinted with permission from You et al…”

Section: Translational Potential Of Iv‐mpm For Cancermentioning

confidence: 99%

Section: Translational Potential Of Iv‐mpm For Cancermentioning

confidence: 99%

Section: Translational Potential Of Iv‐mpm For Cancermentioning

confidence: 99%

Section: Monitoring Chemotherapy Distribution Effects and Resistamentioning

confidence: 99%

Section: Label-free Iv-mpmmentioning

confidence: 99%

See 3 more Smart Citations

Frontiers in intravital multiphoton microscopy of cancer

2019

View full text Add to dashboard Cite

show abstract

Nonlinear Spectral‐Imaging Study of Second‐ and Third‐Harmonic Enhancements by Surface‐Lattice Resonances

Shen

Liu

et al. 2020

Advanced Optical Materials

View full text Add to dashboard Cite

the collective charge density scattered by a single electron. Hence, the dephasing time significantly affects the field-enhanced intensity of surface plasmon resonance, as well as the intensity magnitude of the nonlinear effect, such as surfaceenhanced Raman scattering, and secondand third-harmonic generation (SHG and THG). [5][6][7][8][9][10][11] To maintain coherent persistence, LSPR can be promoted to surface lattice resonance (SLR) by constructing a repetitive array with a periodicity approaching the LSPR wavelength of nanoparticles, which produces leptokurtic spectral features (referred to as Wood or Rayleigh anomalies). [12][13][14][15] Therefore, in addition to the shape and size of the nanoparticles, the shape and periodic parameters of the lattice inevitably influence the resonance mass and intensity. Highly narrow resonances can be achieved by tuning the lattice constant within manufacturing technology, which has been studied extensively. [15][16][17] However, the surface nonlinearity of two-photon excited fluorescence (2PEF), three-photon excited fluorescence (3PEF), SHG, and THG, which is affected by the lattice resonance, and has been revealed by reflected spectral and inverted microscopic methods, particularly with high sensitivity and high resolution, remains insufficiently understood. The vibration modes of SLR, driven by near infrared (NIR) I (750-1000 nm) or NIR II (1000-1700 nm) lasers on different nanoparticle arrays and significantly enhancing the local fields at the particles, require further exploration. Moreover, employing diverse excitation wavelengths will result in chromatic aberration and resolution reduction, which can be improved by technological means and computational solutions, such as the design of higher NA objectives, localization and tracking of individual fluorescent particles and molecules, [18,19] and modeling of a reliable point spread function. [20] Although saturated structured-illumination microscopy, [21] photoactivated localization microscopy, [22,23] stimulated emission depletion microscopy, [24,25] and multiphoton harmonic spatial frequency modulation imaging [26][27][28] have made tremendous inroads toward approaching or breaking the diffraction limit for multiphoton microscopy, a lack of the determination of optical resolutions for the nonlinear optical imaging of metasurfaces remains. The resolution that can be achieved by a system determines whether it can reveal the lattice resonance on the nanoscale.An ultrafast spectral imaging optical system featuring adaptive precompensation for group delay dispersion, automated tuning and collimation, rapid power stabilization, and rapid spectral imaging switching is developed. In this system, real-time visualization ensures that the excited Gaussian beam remains focused on the nanoarray of the electron-beam lithographed metasurface, as well as maximum gain of the nonlinear effect information. Based on the system, two surface-lattice resonance metasurfaces under broadly tunable excitation (800-1300 nm) are investigated. Th...

show abstract

Quantitative Third Harmonic Generation Microscopy for Assessment of Glioma in Human Brain Tissue

et al. 2019

View full text Add to dashboard Cite

Distinguishing tumors from normal brain cells is important but challenging in glioma surgery due to the lack of clear interfaces between the two. The ability of label‐free third harmonic generation (THG) microscopy in combination with automated image analysis to quantitatively detect glioma infiltration in fresh, unprocessed tissue in real time is assessed. The THG images reveal increased cellularity in grades II–IV glioma samples from 23 patients, as confirmed by subsequent hematoxylin and eosin histology. An automated image quantification workflow is presented for quantitative assessment of the imaged cellularity as a reflection of the degree of glioma invasion. The cellularity is validated in three ways: 1) Quantitative comparison of THG imaging with fluorescence microscopy of nucleus‐stained samples demonstrates that THG reflects the true tissue cellularity. 2) Thresholding of THG cellularity differentiates normal brain from glioma infiltration, with 96.6% sensitivity and 95.5% specificity, in nearly perfect (93%) agreement with pathologists. 3) In one patient, a good correlation between THG cellularity and preoperative magnetic resonance and positron emission tomography imaging is demonstrated. In conclusion, quantitative real‐time THG microscopy accurately assesses glioma infiltration in ex vivo human brain samples, and therefore holds strong potential for improving the accuracy of surgical resection.

show abstract

Intravital imaging by simultaneous label-free autofluorescence-multiharmonic microscopy

Cited by 222 publications

References 59 publications

Frontiers in intravital multiphoton microscopy of cancer

Frontiers in intravital multiphoton microscopy of cancer

Nonlinear Spectral‐Imaging Study of Second‐ and Third‐Harmonic Enhancements by Surface‐Lattice Resonances

Quantitative Third Harmonic Generation Microscopy for Assessment of Glioma in Human Brain Tissue

Contact Info

Product

Resources

About