CLARITY with neuronal tracing and immunofluorescence to study the somatosensory system in rats

Martínez‐Lorenzana, Guadalupe; Gamal-Eltrabily, Mohammed; Tello-García, Irma Alejandra; A, Martínez Torres; Becerra-González, Marymar; González‐Hernández, Abimael; Condés‐Lara, Miguel

doi:10.1016/j.jneumeth.2020.109048

Cited by 4 publications

(3 citation statements)

References 41 publications

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“…Meningeal tissue was treated using CLARITY ( 14 ); optical microscopy and digital image processing were performed using a home-made setup ( 15 ) and Fiji ( 16 ), respectively.…”

Section: Methodsmentioning

confidence: 99%

Optical sectioning of unlabeled samples using bright-field microscopy

Gutiérrez–Medina

2022

Proc. Natl. Acad. Sci. U.S.A.

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The bright-field (BF) optical microscope is a traditional bioimaging tool that has been recently tested for depth discrimination during evaluation of specimen morphology; however, existing approaches require dedicated instrumentation or extensive computer modeling. We report a direct method for three-dimensional (3D) imaging in BF microscopy, applicable to label-free samples, where we use Köhler illumination in the coherent regime and conventional digital image processing filters to achieve optical sectioning. By visualizing fungal, animal tissue, and plant samples and comparing with light-sheet fluorescence microscopy imaging, we demonstrate the accuracy and applicability of the method, showing how the standard microscope is an effective 3D imaging device.

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“…Meningeal tissue was treated using CLARITY ( 14 ); optical microscopy and digital image processing were performed using a home-made setup ( 15 ) and Fiji ( 16 ), respectively.…”

Section: Methodsmentioning

confidence: 99%

Optical sectioning of unlabeled samples using bright-field microscopy

Gutiérrez–Medina

2022

Proc. Natl. Acad. Sci. U.S.A.

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“…To perform block staining, it was possible to use the same staining solution used for sections simply by extending the staining duration in the present study. Many methods of immunostaining tissues have been reported along with the clearing method (Belle et al, 2014;Du et al, 2019;Kubota et al, 2017;Lee et al, 2016;Martinez-Lorenzana et al, 2021;Renier et al, 2014), and some even label RNA within the clearing tissues (Sylwestrak et al, 2016;Wang et al, 2018a;Ye et al, 2016). However, the appropriate conditions must be precisely determined for almost every antibody.…”

Section: Tissue Staining and Prevention Of Fadingmentioning

confidence: 99%

biPACT: A method for three-dimensional visualization of mouse spinal cord circuits of long segments with high resolution

Nakanishi

Shinozaki

Nagoshi

et al. 2022

Journal of Neuroscience Methods

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“…Tissue-clearing techniques emerged in the last decade to allow high-resolution 3D imaging of biological tissues. Numerous tissue clearing methods are currently available such as DISCO (iDISCO, uDISCO and 3DISCO) [37,40,41], CLARITY [34,[42][43][44][45][46][47], and seeDB [39], among others. Most of these protocols reduce the light scattering provoked by the presence of the lipid and homogenize the RI, obtaining more transparent tissues [48].…”

Section: Introductionmentioning

confidence: 99%

An Image-Based Framework for the Analysis of the Murine Microvasculature: From Tissue Clarification to Computational Hemodynamics

et al. 2022

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The blood–brain barrier is a unique physiological structure acting as a filter for every molecule reaching the brain through the blood. For this reason, an effective pharmacologic treatment supplied to a patient by systemic circulation should first be capable of crossing the barrier. Standard cell cultures (or those based on microfluidic devices) and animal models have been used to study the human blood–brain barrier. Unfortunately, these tools have not yet reached a state of maturity because of the complexity of this physiological process aggravated by a high heterogeneity that is not easily recapitulated experimentally. In fact, the extensive research that has been performed and the preclinical trials carried out provided sometimes contradictory results, and the functionality of the barrier function is still not fully understood. In this study, we have combined tissue clarification, advanced microscopy and image analysis to develop a one-dimensional computational model of the microvasculature hemodynamics inside the mouse brain. This model can provide information about the flow regime, the pressure field and the wall shear stress among other fluid dynamics variables inside the barrier. Although it is a simplified model of the cerebral microvasculature, it allows a first insight on into the blood–brain barrier hemodynamics and offers several additional possibilities to systematically study the barrier microcirculatory processes.

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CLARITY with neuronal tracing and immunofluorescence to study the somatosensory system in rats

Cited by 4 publications

References 41 publications

Optical sectioning of unlabeled samples using bright-field microscopy

Optical sectioning of unlabeled samples using bright-field microscopy

biPACT: A method for three-dimensional visualization of mouse spinal cord circuits of long segments with high resolution

An Image-Based Framework for the Analysis of the Murine Microvasculature: From Tissue Clarification to Computational Hemodynamics

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