Significance: Histopathological analysis of tissues is an essential tool for grading, staging, diagnosing and resecting cancers and other malignancies. Current histopathological 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 novel 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 of the PAARS microscope takes advantage of DNA's unique optical absorption to provide nuclear contrast analogous to hematoxylin staining of cell nucelli. Concurrently, the optical scattering contrast of the PARS detection system is leveraged to provide bulk tissue contrast analogous to 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 extra-nuclear features such as cancerous cells, glands and ducts, adipocytes, and stromal structures such as collagen.
Conclusions. The presented dual contrast PARS microscope enables label-free visualizations of tissue with contrast and quality analogous 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.
Histological images are critical in the diagnosis and treatment of cancers. Unfortunately, the current method for capturing these microscopy images require resource intensive tissue preparation that delays diagnosis for many days to a few weeks. To streamline this process, clinicians are limited to assessing small macroscopically representative subsets of tissues. Here, we present a combined photoacoustic remote sensing (PARS) microscope and swept source optical coherence tomography (SS-OCT) system designed to circumvent these diagnostic limitations. The proposed multimodal microscope provides…
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 reflection-mode imaging modality that reveals cellular-scale morphology by detecting intrinsic endogenous contrast. We accomplish this with…
Histopathological visualizations are a pillar of modern medicine and biological research. Surgical oncology relies exclusively on post-operative histology to determine definitive surgical success and guide adjuvant treatments. The current histology workflow is based on bright-field microscopic assessment of histochemical stained tissues and has some major limitations. For example, the preparation of stained specimens for brightfield assessment requires lengthy sample processing, delaying interventions for days or even weeks. Hence, there is a pressing need for improved histopathology methods. …
Photoacoustic computed tomography (PACT) combines the optical contrast of optical imaging and the penetrability of sonography. In this work, we develop a novel PACT system to provide real-time imaging, which is achieved by a 120-elements ultrasound array only using a single data acquisition (DAQ) channel. To reduce the channel number of DAQ, we superimpose 30 nearby channels' signals together in the analog domain, and shrinking to 4 channels of data (120/30=4). Furthermore, a four-to-one delay-line module is designed to combine these four channels' data into one channel before entering the sin…
Photoacoustic imaging has emerged in the past decades. Compared with the traditional medical imaging mode, it has better imaging performance and has great development potential in the field of biological imaging. In traditional photoacoustic microscopy, a single laser pulse is generally used to irradiate the sample to produce photoacoustic signal. And signal-to-noise ratio (SNR) is a very important indicator for photoacoustic imaging. In order to obtain the image with high SNR, multiple acquisition or increasing laser pulse energy is usually adopted. The former will lead to slower imaging spee…
Photoacoustic Computed Tomography (PACT) is a major configuration of photoacoustic imaging, a hybrid noninvasive modality for both functional and molecular imaging. PACT has rapidly gained importance in the field of biomedical imaging due to superior performance as compared to conventional optical imaging counterparts. However, the overall cost of developing a PACT system is one of the challenges towards clinical translation of this novel technique. The cost of a typical commercial PACT system originates from optical source, ultrasound detector, and data acquisition unit. With growing applicat…
In the past decades, absorption modalities have emerged as powerful tools for label-free functional and structural imaging of cells and tissues. Many biomolecules present unique absorption spectra providing chromophore-specific information on properties such as chemical bonding, and sample composition. As chromophores absorb photons the absorbed energy is emitted as photons (radiative relaxation) or converted to heat and under specific conditions pressure (non-radiative relaxation). Modalities like fluorescence microscopy may capture radiative relaxation to provide contrast, while modalities l…
Photoacoustic Remote Sensing (PARS) microscopy is an emerging label-free optical absorption imaging modality. PARS operates by capturing nanosecond-scale optical perturbations generated by photoacoustic pressures. These time-domain (TD) modulations are usually projected by amplitude to determine absorption magnitude. However, significant information on the target's material properties is contained within the TD signals. This work proposes a novel clustering method to learn TD features which relate to underlying biomolecule characteristics. This technique identifies features related to constitu…
As an emerging medical diagnostic technology, photoacoustic imaging has been implemented for both preclinical and clinical applications. For clinical convenience, a handheld free scan photoacoustic tomography (PAT) system providing 3D imaging capability is essentially needed, which has potential for surgical navigation and disease diagnosis. In this paper, we proposed a free scan 3D PAT (fsPAT) system based on a handheld linear array ultrasound probe. A global positioning system (GPS) is applied for ultrasound probes coordinate acquisition. The proposed fsPAT can simultaneously realize real ti…
Photoacoustic imaging can achieve high-resolution three-dimensional visualization of optical absorbers at penetration depths ~ 1 cm in biological tissues by detecting optically-induced high ultrasound frequencies. Tomographic acquisition with ultrasound linear arrays offers an easy implementation of single-side access, parallelized and high-frequency detection, but usually comes with an image quality impaired by the directionality of the detectors. Indeed, a simple translation of the array perpendicularly to its median imaging plane is often used, but results both in a poor resolution in the t…
German Cancer Research Center（德国癌症研究中心）；Heidelberg University（海德堡大学）；University of Bern（伯尔尼大学）
Multispectral photoacoustic imaging (PAI) is an emerging imaging modality which enables the recovery of functional tissue parameters such as blood oxygenation. However, the underlying inverse problems are potentially ill-posed, meaning that radically different tissue properties may - in theory - yield comparable measurements. In this work, we present a new approach for handling this specific type of uncertainty by leveraging the concept of conditional invertible neural networks (cINNs). Specifically, we propose going beyond commonly used point estimates for tissue oxygenation and converting si…
German Cancer Research Center (DKFZ)（德国癌症研究中心（DKFZ））；Heidelberg University（海德堡大学）；Heidelberg University Hospital（海德堡大学医院）
Purpose: Photoacoustic tomography (PAT) is a novel imaging technique that can spatially resolve both morphological and functional tissue properties, such as the vessel topology and tissue oxygenation. While this capacity makes PAT a promising modality for the diagnosis, treatment and follow-up of various diseases, a current drawback is the limited field-of-view (FoV) provided by the conventionally applied 2D probes. Methods: In this paper, we present a novel approach to 3D reconstruction of PAT data (Tattoo tomography) that does not require an external tracking system and can smoothly be integ…
Photoacoustic imaging of human brain function through intact adult skulls
California Institute of Technology（加州理工学院）；Case Western Reserve University（凯斯西储大学）；University of Southern California（南加州大学）；Midjourney, Inc.（Midjourney公司）；Changsha University of Science and Technology（长沙理工大学）；Peking University（北京大学）；European Molecular Biology Laboratory（欧洲分子生物学实验室）；Rice University（莱斯大学）；Tsinghua University（清华大学）
University of California, Los Angeles（加州大学洛杉矶分校）；California NanoSystems Institute (CNSI)（加州纳米系统研究所）；David Geffen School of Medicine（戴维·格芬医学院）
Pathological diagnosis relies on the visual inspection of histologically stained thin tissue specimens, where different types of stains are applied to bring contrast to and highlight various desired histological features. However, the destructive histochemical staining procedures are usually irreversible, making it very difficult to obtain multiple stains on the same tissue section. Here, we demonstrate a virtual stain transfer framework via a cascaded deep neural network (C-DNN) to digitally transform hematoxylin and eosin (H&E) stained tissue images into other types of histological stains. U…
Spectral micro-CT for quantitative analysis of calcification in fibrocartilage
INFN - Division of Ferrara（意大利国家核物理研究所费拉拉分部）；Dept. of Physics and Earth Science, University of Ferrara（费拉拉大学地球科学与物理学系）；Dept. of Physics, University of Trieste（的里雅斯特大学物理系）；INFN - Division of Trieste（意大利国家核物理研究所的里雅斯特分部）；RAMSES Laboratory, RIT Department, IRCCS Istituto Ortopedico Rizzoli（IRCCS里佐利骨科医院RIT部门RAMSES实验室）；nd Orthopaedic and Traumatologic Clinic, IRCCS Istituto Ortopedico Rizzoli（IRCCS里佐利骨科医院第二骨科与创伤科诊所）
Fourier ptychographic microscopy (FPM) is a computational approach geared towards creating high-resolution and large field-of-view images without mechanical scanning. To acquire color images of histology slides, it often requires sequential acquisitions with red, green, and blue illuminations. The color reconstructions often suffer from coherent artifacts that are not presented in regular incoherent microscopy images. As a result, it remains a challenge to employ FPM for digital pathology applications, where resolution and color accuracy are of critical importance. Here we report a deep learni…
