Ultraviolet photoacoustic remote sensing microscopy

Haven, Nathaniel J. M.; Bell, Kevan; Kedarisetti, Pradyumna; Lewis, John D.; Zemp, Roger J.

doi:10.1364/ol.44.003586

Cited by 40 publications

(31 citation statements)

References 22 publications

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“…These capabilities, as highlighted earlier, lead to a device suitable for intraoperative and clinical environments. Previously, PARS has demonstrated the ability to visualize nuclear contrast in chicken embryos and animal tissue 32,33 . However, these reports were limited in their field of view and did not reveal significant structural information.…”

Section: Introductionmentioning

confidence: 99%

Non-contact reflection-mode optical absorption spectroscopy using photoacoustic remote sensing

Bell

Reza

2020

Opt. Lett.

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Histological visualizations are critical to clinical disease management and are fundamental to biological understanding. However, current approaches that rely on bright-field microscopy require extensive tissue preparation prior to imaging. These processes are labor intensive and contribute to delays in clinical feedback that can extend to two to three weeks for standard paraffin-embedded tissue preparation and interpretation. Here, we present a label-free reflectionmode imaging modality that reveals cellular-scale morphology by detecting intrinsic endogenous contrast. We accomplish this with the novel photoacoustic remote sensing (PARS) detection system that permits non-contact optical absorption contrast to be extracted from thick and opaque biological targets with optical resolution. PARS was examined both as a rapid assessment tool that is capable of managing large samples (>1 cm 2) in under 10 minutes, and as a high contrast imaging modality capable of extracting specific biological contrast to simulate conventional histological stains such as hematoxylin and eosin (H&E). The capabilities of the proposed method are demonstrated in a variety of human tissue preparations including formalin-fixed paraffinembedded tissue blocks and unstained slides sectioned from these blocks, including normal and neoplastic human brain, and breast epithelium involved with breast cancer. Similarly, PARS images of human skin prepared by frozen section clearly demonstrated basal cell carcinoma and normal human skin tissue. Finally, we imaged unprocessed murine kidney and achieved histologically relevant subcellular morphology in fresh tissue. This represents a vital step towards an effective real-time clinical microscope that overcomes the limitations of standard histopathologic tissue preparations and enables real-time pathology assessment.

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Section: Introductionmentioning

confidence: 99%

Non-contact reflection-mode optical absorption spectroscopy using photoacoustic remote sensing

Bell

Reza

2020

Opt. Lett.

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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%

“… 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%

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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“…Compared to the interferometric method, the non-interferometric approach extracts PA signals by measuring the intensity change of the back-reflected probing light source induced by the initial pressure ( Fig. 7 c) [ [137] , [138] , [139] , [140] ]. In this method, the probing and the excitation light sources are co-axially focused on generating the strongest pressure.…”

Section: Fast Photoacoustic Microscopy As a Function Of Light Source Speedmentioning

confidence: 99%

High-speed photoacoustic microscopy: A review dedicated on light sources

Cho

Park

et al. 2021

Photoacoustics

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In recent years, many methods have been investigated to improve imaging speed in photoacoustic microscopy (PAM). These methods mainly focused upon three critical factors contributing to fast PAM: laser pulse repetition rate, scanning speed, and computing power of the microprocessors. A high laser repetition rate is fundamentally the most crucial factor to increase the PAM speed. In this paper, we review methods adopted for fast PAM systems in detail, specifically with respect to light sources. To the best of our knowledge, ours is the first review article analyzing the fundamental requirements for developing high-speed PAM and their limitations from the perspective of light sources.

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Ultraviolet photoacoustic remote sensing microscopy

Cited by 40 publications

References 22 publications

Non-contact reflection-mode optical absorption spectroscopy using photoacoustic remote sensing

Non-contact reflection-mode optical absorption spectroscopy using photoacoustic remote sensing

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

High-speed photoacoustic microscopy: A review dedicated on light sources

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