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

Adams, Wilson R.; Mehl, Brian P.; Lieser, Eric; Wang, Manqing; Patton, Shane; Throckmorton, Graham A.; Jenkins, J. Logan; Ford, Jeremy B.; Gautam, Rekha; Brooker, Jeff Z.; Jansen, E.; Mahadevan‐Jansen, Anita

doi:10.1101/2020.04.06.023820

Cited by 6 publications

(8 citation statements)

References 80 publications

(124 reference statements)

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“…Following confirmation of our instrument’s ability to obtain hsSRS image stacks from live NG108 cells ( Figure 1D and E ), initial experiments with IR stimulation during nonlinear microscopy (i.e. any coherent Raman modality, multiphoton fluorescence, or higher harmonic generation) resulted in a substantial loss in measured signal during IR exposure ( Figure 2B and C ) (22). This was apparent in both short periods of heating from a millisecond pulse of IR light (unpublished data), or pulse trains of multiple microsecond pulses of light.…”

Section: Resultsmentioning

confidence: 75%

“…Furthermore, coherent Raman imaging can be readily performed simultaneously with other nonlinear microscopy modalities (22). Multiplexing modalities might enable studies into how lipid membrane biophysics can influence biological dynamics with conventionally accepted molecular reporters.…”

Section: Discussionmentioning

confidence: 99%

“…The physical layout and capability of the custom-built multimodal imaging platform utilized in this study ( Figure 1A ) was described previously (22). Briefly, a dual output femtosecond near-infrared laser source (Insight DS+, Spectra Physics, Santa Clara, CA) was used to excite nonlinear contrast.…”

Section: Methodsmentioning

confidence: 99%

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Visualizing the Role of Lipid Dynamics during Infrared Neural Stimulation with Hyperspectral Stimulated Raman Scattering Microscopy

Adams

Gautam

Locke

et al. 2021

Preprint

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Infrared neural stimulation, or INS, is a method of using pulsed infrared light to yield label-free neuronal stimulation with broad experimental and translational utility. Despite its robust demonstration, the mechanistic and biophysical underpinnings of INS have been the subject of debate for more than a decade. The role of lipid membrane thermodynamics appears to play an important role in how fast IR-mediated heating nonspecifically drives action potential generation. Direct observation of lipid membrane dynamics during INS remains to be shown in a live neural model system. To directly test the involvement of lipid dynamics in INS, we used hyperspectral stimulated Raman scattering (hsSRS) microscopy to study biochemical signatures of high speed vibrational dynamics underlying INS in a live neural cell culture model. Findings suggest that lipid bilayer structural changes are occurring during INS in vitro in NG108-15 neuroglioma cells. Lipid-specific signatures of cell SRS spectra were found to vary with stimulation energy and radiant exposure. Spectroscopic observations were verified against high-speed ratiometric fluorescence imaging of a conventional lipophilic membrane structure reporter, di-4-ANNEPS. Overall, the presented data supports the hypothesis that INS causes changes in the lipid membrane of neural cells by changing lipid membrane packing order - which coincides with likelihood of cell stimulation. Furthermore, this work highlights the potential of hsSRS as a method to study biophysical and biochemical dynamics safely in live cells.

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Section: Resultsmentioning

confidence: 75%

Section: Discussionmentioning

confidence: 99%

See 1 more Smart Citation

Visualizing the Role of Lipid Dynamics during Infrared Neural Stimulation with Hyperspectral Stimulated Raman Scattering Microscopy

Adams

Gautam

Locke

et al. 2021

Preprint

Self Cite

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show abstract

“…From the above discussion, no single imaging technology can present a global understanding to the complex biological structure and process. Therefore, multi-technique combination is a trend for imaging and information analysis to complicated samples [124][125][126]. New imaging schemes will make these label-free laser scanning microscopies to be more powerful biological imaging tool when combine with spectroscopy, laser and detection technology.…”

Section: Discussionmentioning

confidence: 99%

Advanced Label-Free Laser Scanning Microscopy and Its Biological Imaging Application

Wang

Huang

2021

Applied Sciences

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By eliminating the photodamage and photobleaching induced by high intensity laser and fluorescent molecular, the label-free laser scanning microscopy shows powerful capability for imaging and dynamic tracing to biological tissues and cells. In this review, three types of label-free laser scanning microscopies: laser scanning coherent Raman scattering microscopy, second harmonic generation microscopy and scanning localized surface plasmon microscopy are discussed with their fundamentals, features and recent progress. The applications of label-free biological imaging of these laser scanning microscopies are also introduced. Finally, the performance of the microscopies is compared and the limitation and perspectives are summarized.

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“…The aforementioned PAM and nonlinear microscopy play an indispensable role in non-invasive labelfree characterization of various biological structures, through either absorption-induced thermoelastic expansion (PAM), intrinsic autofluorescence (MPM), molecular vibration (SRS), or non-centrosymmetric orientation (SHG). The simultaneous multiplexing 21,22 of these methods enables cell phenotyping and classification based on the targeted biomolecules. However, the low-throughput nature of these scanningbased imaging modalities ultimately hinders their clinical translations.…”

Section: Introductionmentioning

confidence: 99%

High-throughput, label-free and slide-free histological imaging by computational microscopy and unsupervised learning

Zhang

Kang

et al. 2021

Preprint

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Rapid and high-resolution histological imaging with minimal tissue preparation has long been a challenging and yet captivating medical pursue. Here, we propose a promising and transformative histological imaging method, termed computational high-throughput autofluorescence microscopy by pattern illumination (CHAMP). With the assistance of computational microscopy, CHAMP enables high-throughput and label-free imaging of thick and unprocessed tissues with large surface irregularity at an acquisition speed of 10 mm2/10 seconds with 1.1-um lateral resolution. Moreover, the CHAMP image can be transformed into a virtually stained histological image (Deep-CHAMP) through unsupervised learning within 15 seconds, where significant cellular features are quantitatively extracted with high accuracy. The versatility of CHAMP is experimentally demonstrated using mouse brain/kidney tissues prepared with various clinical protocols, which enables a rapid and accurate intraoperative/postoperative pathological examination without tissue processing or staining, demonstrating its great potential as an assistive imaging platform for surgeons and pathologists to provide optimal adjuvant treatment.

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

Cited by 6 publications

References 80 publications

Visualizing the Role of Lipid Dynamics during Infrared Neural Stimulation with Hyperspectral Stimulated Raman Scattering Microscopy

Visualizing the Role of Lipid Dynamics during Infrared Neural Stimulation with Hyperspectral Stimulated Raman Scattering Microscopy

Advanced Label-Free Laser Scanning Microscopy and Its Biological Imaging Application

High-throughput, label-free and slide-free histological imaging by computational microscopy and unsupervised learning

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