Optical clearing in cardiac imaging: A comparative study

Olianti, Camilla; Giardini, Francesco; Lazzeri, Erica; Costantini, Irene; Silvestri, Ludovico; Coppini, Raffaele; Cerbai, Elisabetta; Pavone, Francesco S.; Sacconi, Leonardo

doi:10.1016/j.pbiomolbio.2021.07.012

Cited by 14 publications

(14 citation statements)

References 30 publications

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“…Interestingly, we observed that the fluorescence of tdTomato was preserved immediately after the completion of the s-DISCO and ECi protocols by decreasing the dehydration time when clearing 100 μm thin lung sections. This finding might explain the variability in the results between users of the same protocols for different applications and samples (40). Moreover, it indicates that optimizing the duration of the dehydration steps might be necessary when implementing a solvent-based optical tissue clearing method where fluorescent proteins are involved.…”

Section: Discussionmentioning

confidence: 99%

Comparison between optical tissue clearing methods for detecting administered mesenchymal stromal cells in mouse lungs

Pichardo

Amadeo

Wilm

et al. 2022

Preprint

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Optical tissue clearing of lung tissue enables the intact lung to be imaged using fluorescence microscopy. Several clearing protocols have been developed in recent years, including the Clear, Unobstructed Brain/Body Imaging Cocktails and Computational analysis (CUBIC), stabilised 3D imaging of solvent-cleared organs (s-DISCO) and Ethyl cinnamate (ECi) methods. Here we compared these protocols with the aim of determining the biodistribution of mesenchymal stroma cells (MSCs) and understanding how they interact with host cells in the mouse lung. First, we evaluated how each method affected the size, morphology, and transparency of the lungs. Then, we compared the preservation of the fluorescence of the protein tdTomato expressed by the MSCs, and of the organic dye Evans Blue which labels the vasculature. In addition, we tested the compatibility of the methods with immunofluorescence staining. We found that CUBIC clearing is the only method that enables direct imaging of fluorescently labelled MSCs in the lungs thereby allowing the study of the MSC interaction with endothelial and immune cells when combined with immunofluorescence staining. Overall, 3D imaging of CUBIC cleared lungs confirmed that injected MSCs are initially retained in the pulmonary microvasculature, and that most cells are eliminated from the lungs within the first 24 h.

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Section: Discussionmentioning

confidence: 99%

Comparison between optical tissue clearing methods for detecting administered mesenchymal stromal cells in mouse lungs

Pichardo

Amadeo

Wilm

et al. 2022

Preprint

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“…Most of them, however, present several limitations such as tissue shrinkage, structural alteration, fluorescence quenching and incompatibility with immunostaining (Silvestri et al, 2016). The challenge of producing large, transparent and fluorescently-labeled volumes has been overcome recently by applying a tissue transformation approach, CLARITY, and it's related derivatives (Chung et al, 2013;Pianca et al, 2019;Olianti et al, 2020Olianti et al, , 2021. These methods allow high transparency, immunolabeling and structural and molecular preservation.…”

Section: Light-sheet Microscopy For Mesoscale Structural Imagingmentioning

confidence: 99%

Novel Optics-Based Approaches for Cardiac Electrophysiology: A Review

et al. 2021

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Optical techniques for recording and manipulating cellular electrophysiology have advanced rapidly in just a few decades. These developments allow for the analysis of cardiac cellular dynamics at multiple scales while largely overcoming the drawbacks associated with the use of electrodes. The recent advent of optogenetics opens up new possibilities for regional and tissue-level electrophysiological control and hold promise for future novel clinical applications. This article, which emerged from the international NOTICE workshop in 20181, reviews the state-of-the-art optical techniques used for cardiac electrophysiological research and the underlying biophysics. The design and performance of optical reporters and optogenetic actuators are reviewed along with limitations of current probes. The physics of light interaction with cardiac tissue is detailed and associated challenges with the use of optical sensors and actuators are presented. Case studies include the use of fluorescence recovery after photobleaching and super-resolution microscopy to explore the micro-structure of cardiac cells and a review of two photon and light sheet technologies applied to cardiac tissue. The emergence of cardiac optogenetics is reviewed and the current work exploring the potential clinical use of optogenetics is also described. Approaches which combine optogenetic manipulation and optical voltage measurement are discussed, in terms of platforms that allow real-time manipulation of whole heart electrophysiology in open and closed-loop systems to study optimal ways to terminate spiral arrhythmias. The design and operation of optics-based approaches that allow high-throughput cardiac electrophysiological assays is presented. Finally, emerging techniques of photo-acoustic imaging and stress sensors are described along with strategies for future development and establishment of these techniques in mainstream electrophysiological research.

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“…Hence, to analyze larger volumes, serial sectioning of the tissue is mandatory. One possibility to reduce the self absorption and scattering of the emitted photons within the sample is tissue clearing [80][81][82]. During this process the refractive index of a chemically fixed tissue is homogenized so that it turns transparent for visible light.…”

Section: Introductionmentioning

confidence: 99%

“…The preparation usually includes decalcification, decolorization, delipidation, dehydration and clearing of the sample. In combination with fluorescent staining and light sheet microscopy, larger volumes in the range of the size of murine brain, a zebra fish or even an entire mouse can be analyzed [81][82][83][84]. However, this technique is also limited by diffraction and has an anisotropic resolution due to the properties of the excitation laser.…”

Section: Introductionmentioning

confidence: 99%

Multiscale X-ray Structural Analysis of Cardiac Cells and Tissues

Reichardt¹

2022

Göttingen Series in X-Ray Physics

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List of publications Danksagung Curriculum Vitaeday. Considering an average volume of 70 ml to 80 ml per beat, more than 7 000 liters of blood are pumped through the body each day. The synchronized contraction of the excitable cardiac tissue cells is controlled by the conduction system, which initiates an electric signal in the sinoatrial node and coordinates the propagation of the action potential through the heart. The continuous function of the heart during its entire lifetime is essential for the proper maintenance of all vital organs. Small dysfunctions or abnormalities can cause drastic changes and effects in the human organism. Therefore, it is very important to understand the cardiac structure from the molecular level up to the scale of the entire organ, as well as its function within the body.The primary aim of this thesis is to develop imaging protocols to probe the entire cy- Cardiac structure and (dys)functionCardiovascular diseases (CVDs) are the main cause of death worldwide with a rising incidence. While CVDs were responsible for less than 10% of the deaths worldwide by the beginning of the 20th century, the number of CVD-related deaths increased to over 30% by 2016 [6-8]. This corresponds to 17.9 million global deaths due to CVDs in one year. The major share of these death (85%) is attributed to heart attacks and from the ancient Egyptians [17]. (B) Drawing of the cardiac anatomy by Aristotle [15]. (C) Sketch of the heart by Leonardo da Vinci [15]. (D) Cardiac model by Richard Lower [18]. (E) Text book figure of Henry Gray's anatomy [16]. (F) Concept of a "Triebwerk" by Ludolf Krehl [18]. (G) Jane Sands Robb's model of the heart [18]. (H) Cardiac fiber model of Alfred Benninghoff and Kurt Goerttler [18]. (I) Model of the helical ventricular band by Francisco Torrent-Guasp [18]. Anatomie des Menschen"), Xavier Bichat ("Anatomie Generale") and Henry Gray and Henry Vandyke Carter ("Grays Anatomy"). Nevertheless, due to ongoing research and new discoveries the textbooks had to be updated constantly. In October 2020, the 42th edition of "Grays Anatomy" was released. Concerning the cardiac anatomy, most of the open questions were related to the cardiac function and structural changes caused by CVDs. For example, in 1891 Ludolf Krehl introduced the idea of a circumferential Historically, the observation of the heart structure started by documenting visual impressions gained from evisceration of animals or autopsies. With progress in the manufacture of optics and the invention of the microscope in the 17th century, it was possible to investigate structures in the micrometer range [48]. During this time, Marcello Malpighi was the first who identified small capillaries in the lungs of bats and established the process of oxygen uptake [49]. This procedure, later called histology (from Greek: histos, tissue and logos, study), revealed its full potential during the 19th Recently, this invention was further developed by a combination of STED and single molecule localization microscopy such as stochastic optical...

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Optical clearing in cardiac imaging: A comparative study

Cited by 14 publications

References 30 publications

Comparison between optical tissue clearing methods for detecting administered mesenchymal stromal cells in mouse lungs

Comparison between optical tissue clearing methods for detecting administered mesenchymal stromal cells in mouse lungs

Novel Optics-Based Approaches for Cardiac Electrophysiology: A Review

Multiscale X-ray Structural Analysis of Cardiac Cells and Tissues

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