Deep tissue imaging: a review from a preclinical cancer research perspective

Feuchtinger, Annette; Walch, Axel; Dobosz, Michael

doi:10.1007/s00418-016-1495-7

Cited by 56 publications

(59 citation statements)

References 74 publications

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“…In recent years, tissue clearing has re‐emerged as a powerful tool for bypassing tissue sectioning. This has led to the development of a plethora of tissue clearing protocols that reduce optical scattering by extracting the lipid fraction and/or by refractive index matching thereby yielding full‐thickness tissue transparency . These protocols allow detailed 3D visualization of intact organs by confocal, light‐sheet, or two‐photon microscopy, but the appropriateness of each method is organ‐specific and depends on the aspired goal in terms of clearing capability, preservation of fluorescent reporter protein signal, compatibility with immunolabeling and nuclear staining, morphological tissue integrity, complexity, and cost …”

Section: Introductionmentioning

confidence: 99%

Comparative analysis reveals Ce3D as optimal clearing method for in toto imaging of the mouse intestine

Bossolani

Pintelon

Detrez

et al. 2019

Neurogastroenterology Motil

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Background The intestinal wall has a complex topographical architecture. The multi‐layered network of the enteric nervous system and its intercellular interactions are difficult to map using traditional section‐based or whole‐mount histology. With the advent of optical clearing techniques, it has become feasible to visualize intact tissue and organs in 3D. However, as yet, a gap still needs to be filled in that no in‐depth analysis has been performed yet on the potential of different clearing techniques for the small intestine. Aim The goal of this study was to identify an optimal clearing protocol for in toto imaging of mouse intestinal tissue. Methods Five aqueous‐based clearing protocols (SeeDB2, CUBIC, ScaleS, Ce3D, and UbasM) and four organic reagent‐based clearing protocols (3DISCO, iDISCO+, uDISCO, and Visikol®) were assessed in segments of small intestine from CX3CR1GFP/GFP and wild‐type mice. Following clearing, optical transparency, tissue morphology, green fluorescent protein (GFP) fluorescence retention, and compatibility with (immuno‐)labeling were analyzed. Key results All organic reagent‐based clearing protocols—except for Visikol—rendered tissue highly transparent but led to substantial tissue shrinkage and deformation. Of the aqueous‐based protocols, only Ce3D yielded full‐thickness tissue transparency. In addition, Ce3D displayed excellent GFP retention and preservation of tissue morphology. Conclusions Ce3D emerged as a most efficient protocol for enabling rapid full‐thickness 3D mapping of the mouse intestinal wall.

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Section: Introductionmentioning

confidence: 99%

Comparative analysis reveals Ce3D as optimal clearing method for in toto imaging of the mouse intestine

Bossolani

Pintelon

Detrez

et al. 2019

Neurogastroenterology Motil

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“…The clearing methods were originally developed for processing large samples obtained from animals (e.g., bone, brain, embryo, heart, intestine, kidney, lung, muscle, pancreas, spinal cord, spleen, testis, among others), as already reviewed in detail in (Feuchtinger et al, 2016;Genina et al, 2010 Figure 2; Ariel, 2017;Seo et al, 2016). Due to the osmotic pressure, the water content in the sample (that has low RI ≈ 1.33) will be passively replaced by the clearing solution.…”

Section: Classification Of the Optical Clearing Methodsmentioning

confidence: 99%

Optical clearing methods: An overview of the techniques used for the imaging of 3D spheroids

Costa

Silva

Moreira

et al. 2019

Biotech & Bioengineering

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Spheroids have emerged as in vitro models that reproduce in a great extent the architectural microenvironment found in human tissues. However, the imaging of 3D cell cultures is highly challenging due to its high thickness, which results in a lightscattering phenomenon that limits light penetration. Therefore, several optical clearing methods, widely used in the imaging of animal tissues, have been recently explored to render spheroids with enhanced transparency. These methods are aimed to homogenize the microtissue refractive index (RI) and can be grouped into four different categories, namely (a) simple immersion in an aqueous solution with high RI;(b) delipidation and dehydration followed by RI matching; (c) delipidation and hyperhydration followed by RI matching; and (d) hydrogel embedding followed by delipidation and RI matching. In this review, the main optical clearing methods, their mechanism of action, advantages, and disadvantages are described. Furthermore, the practical examples of the optical clearing methods application for the imaging of 3D spheroids are highlighted. K E Y W O R D Simaging, light scattering, optical clearing, optical microscopy, spheroids.

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“…These results suggest that cCLOT will be an effective method in future studies using freshly drawn human blood or fixed samples of human thrombi. Significant progress has been made in recent years towards developing clearing solutions and protocols to facilitate fluorescence microscopy deeper into tissues [22,23,29,30]. An effort to clear whole mouse body tissues resulted in the development of CUBIC-1, which eluted heme from PFA fixed blood suspensions compatible with fluorescent protein imaging.…”

Section: Application Of Cclot Methods To Human Bloodmentioning

confidence: 99%

Whole blood clot optical clearing for nondestructive 3D imaging and quantitative analysis

Höök¹,

Brito-Robinson²,

Ким³

et al. 2017

Biomed. Opt. Express

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A technological revolution in both light and electron microscopy imaging now allows unprecedented views of clotting, especially in animal models of hemostasis and thrombosis. However, our understanding of three-dimensional high-resolution clot structure remains incomplete since most of our recent knowledge has come from studies of relatively small clots or thrombi, due to the optical impenetrability of clots beyond a few cell layers in depth. Here, we developed an optimized optical clearing method termed cCLOT that renders large whole blood clots transparent and allows confocal imaging as deep as one millimeter inside the clot. We have tested this method by investigating the 3D structure of clots made from reconstituted pre-labeled blood components yielding new information about the effects of clot contraction on erythrocytes. Although it has been shown recently that erythrocytes are compressed to form polyhedrocytes during clot contraction, observations of this phenomenon have been impeded by the inability to easily image inside clots. As an efficient and nondestructive method, cCLOT represents a powerful research tool in studying blood clot structure and mechanisms controlling clot morphology. Additionally, cCLOT optical clearing has the potential to facilitate imaging of ex vivo clots and thrombi derived from healthy or pathological conditions. W. Weisel, I. N. Chernysh, C. Nagaswami, P. R. Williams, and K. Hawkins, "Effects of unidirectional flow shear stresses on the formation, fractal microstructure and rigidity of incipient whole blood clots and fibrin gels," Clin. Hemorheol. Microcirc. 60(4), 451-464 (2015).

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Deep tissue imaging: a review from a preclinical cancer research perspective

Cited by 56 publications

References 74 publications

Comparative analysis reveals Ce3D as optimal clearing method for in toto imaging of the mouse intestine

Comparative analysis reveals Ce3D as optimal clearing method for in toto imaging of the mouse intestine

Optical clearing methods: An overview of the techniques used for the imaging of 3D spheroids

Whole blood clot optical clearing for nondestructive 3D imaging and quantitative analysis

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