In vivo combined virtual histology and vascular imaging with dual-wavelength photoacoustic remote sensing microscopy

Restall, Brendon S.; Haven, Nathaniel J. M.; Kedarisetti, Pradyumna; Zemp, Roger J.

doi:10.1364/osac.398269

Cited by 17 publications

(17 citation statements)

References 28 publications

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“… 15 – 21 These techniques have subsequently provided hematoxylin-like visualizations of nuclear structures in tissue blocks, slides, and fresh tissues. 15 – 21 However, these techniques do not provide full H&E emulation, as they lack the extranuclear tissue structures captured by eosin staining. Here, we propose approaching this problem by leveraging, for the first time, PARS unique dual scattering and absorption contrast.…”

Section: Resultsmentioning

confidence: 99%

“… 14 Moreover, PARS may provide chromophore-specific contrast by selecting excitation wavelengths to target unique biomolecule absorption spectra. 15 – 18 Applied to histological imaging, PARS has successfully used UV excitation to capture nuclear structures. 15 – 21 This technique is efficacious in providing hematoxylin-like contrast in frozen sections and paraffin-embedded blocks and sections.…”

Section: Introductionmentioning

confidence: 99%

“… 15 – 18 Applied to histological imaging, PARS has successfully used UV excitation to capture nuclear structures. 15 – 21 This technique is efficacious in providing hematoxylin-like contrast in frozen sections and paraffin-embedded blocks and sections. 15 – 21 However, these UV-PARS visualizations lack some essential diagnostic details.…”

Section: Introductionmentioning

confidence: 99%

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Single acquisition label-free histology-like imaging with dual-contrast photoacoustic remote sensing microscopy

2021

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Significance: Histopathological analysis of tissues is an essential tool for grading, staging, diagnosing, and resecting cancers and other malignancies. Current histopathological imaging techniques require substantial sample processing, prior to staining with hematoxylin and eosin (H&E) dyes, to highlight nuclear and cellular morphology. Sample preparation and staining is resource intensive and introduces potential for variability during sample preparation.Aim: We present a method for direct label-free histopathological assessment of formalin-fixed paraffin-embedded tissue blocks and thin tissue sections using a dual-contrast photoacoustic remote sensing (PARS) microscopy system.Approach: To emulate the nuclear and cellular contrast of H&E staining, we leverage unique properties of the PARS system. Here, the ultraviolet excitation PARS microscope takes advantage of DNA's unique optical absorption to provide nuclear contrast analogous to hematoxylin staining of cell nuclei. Concurrently, the optical scattering contrast of the PARS detection system is leveraged to provide bulk tissue contrast reminiscent of eosin staining of cell membranes. Results:We demonstrate the efficacy of this technique by imaging human breast tissue and human skin tissues in formalin-fixed paraffin-embedded tissue blocks and frozen sections, respectively. Salient nuclear and extranuclear features such as cancerous cells, glands and ducts, adipocytes, and stromal structures such as collagen are captured. Conclusions:The presented dual-contrast PARS microscope enables label-free visualization of tissues with contrast and quality comparable to the current gold standard for histopathological analysis. Thus, the proposed system is well positioned to augment existing histopathological workflows, providing histological imaging directly on unstained tissue blocks and sections.

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Section: Resultsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Single acquisition label-free histology-like imaging with dual-contrast photoacoustic remote sensing microscopy

2021

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“…This demonstrates that there is a small photobleaching effect as the nuclei are not as prominent post-UV-PARS exposure, but also that there is no excessive damage to the sample itself as the power exposures of 5 and 45 nJ for the 266- and 532-nm light, respectively, are below the American National Standards Institute limits. 23 …”

Section: Resultsmentioning

confidence: 99%

“… 20 – 22 Multicontrast imaging has recently been demonstrated, where multiple pulsed excitation wavelengths are interlaced to generate absorption contrast from several chromophores. 23 – 26 In this paper, we demonstrate a combined contrast imaging system for phantom imaging experiments and further apply the system to cell and tissue imaging applications and to validate the label-free 3D virtual histology capabilities of ultraviolet-PARS (UV-PARS) using fluorescent labeling and confocal microscopy. This is the first report combining optical absorption and confocal fluorescence contrast in a high-resolution, non-contact, optical imaging system.…”

Section: Introductionmentioning

confidence: 99%

Multimodal 3D photoacoustic remote sensing and confocal fluorescence microscopy imaging

et al. 2021

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. Significance: Complementary absorption and fluorescence contrast could prove useful for a wide range of biomedical applications. However, current absorption-based photoacoustic microscopy systems require the ultrasound transducers to physically touch the samples, thereby increasing contamination and limiting strong optical focusing in reflection mode. Aim : We sought to develop an all-optical system for imaging cells and tissues using the three combined imaging modalities: photoacoustic remote sensing (PARS), epifluorescence, and confocal laser scanning microscopy (CLSM). Approach: A PARS subsystem with ultraviolet excitation was used to obtain label-free absorption-contrast images of nucleic acids in ex vivo tissue samples. Co-integrated epifluorescence and CLSM subsystems were used to verify the 2D and 3D nuclei distribution. Results : Complementary absorption and fluorescence contrast were demonstrated in phantom imaging experiments and subsequent cell and tissue imaging experiments. Lateral and axial resolution of ultraviolet-PARS (UV-PARS) is shown to be 0.39 and , respectively, with 266-nm light. CLSM lateral and axial resolution was measured as 0.97 and , respectively. This resolution is sufficient to image individual cell layers with fine optical sectioning. UV-PARS images of cell nuclei are validated in thick tissue using CLSM. Conclusions : Multimodal absorption and fluorescence contrast are obtained with a non-contact all-optical microscopy system for the first time and utilized to obtain images of cells and tissues with subcellular resolution.

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Imaging approaches for monitoring three‐dimensional cell and tissue culture systems

Hilzenrat

Gill

McArthur

2022

Journal of Biophotonics

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The past decade has seen an increasing demand for more complex, reproducible and physiologically relevant tissue cultures that can mimic the structural and biological features of living tissues. Monitoring the viability, development and responses of such tissues in real‐time are challenging due to the complexities of cell culture physical characteristics and the environments in which these cultures need to be maintained in. Significant developments in optics, such as optical manipulation, improved detection and data analysis, have made optical imaging a preferred choice for many three‐dimensional (3D) cell culture monitoring applications. The aim of this review is to discuss the challenges associated with imaging and monitoring 3D tissues and cell culture, and highlight topical label‐free imaging tools that enable bioengineers and biophysicists to non‐invasively characterise engineered living tissues.

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In vivo combined virtual histology and vascular imaging with dual-wavelength photoacoustic remote sensing microscopy

Cited by 17 publications

References 28 publications

Single acquisition label-free histology-like imaging with dual-contrast photoacoustic remote sensing microscopy

Single acquisition label-free histology-like imaging with dual-contrast photoacoustic remote sensing microscopy

Multimodal 3D photoacoustic remote sensing and confocal fluorescence microscopy imaging

Imaging approaches for monitoring three‐dimensional cell and tissue culture systems

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