Whole-body tissue stabilization and selective extractions via tissue-hydrogel hybrids for high-resolution intact circuit mapping and phenotyping

Treweek, Jennifer B.; Chan, Ken Y.; Flytzanis, Nicholas C.; Yang, Baohua; Deverman, Benjamin E.; Greenbaum, Alon; Lignell, Antti; Xia, Cheng; Cai, Long; Ladinsky, Mark S.; Björkman, Pamela J.; Fowlkes, Charless C.; Gradinaru, Viviana

doi:10.1038/nprot.2015.122

Cited by 234 publications

(328 citation statements)

References 204 publications

(225 reference statements)

Supporting

Mentioning

321

Contrasting

Order By: Relevance

“…The relative strengths and weaknesses of optical clearing techniques have been discussed previously in the literature 13,17,22, , but are most clearly laid out in 23 . Passive CLARITY is well suited for experiments where large numbers of samples are being cleared simultaneously and reproducibility is of great importance.…”

Section: Discussionmentioning

confidence: 99%

Optical Clearing of the Mouse Central Nervous System Using Passive CLARITY

Roberts

Johnsonbaugh

Spence

et al. 2016

JoVE

View full text Add to dashboard Cite

Traditionally, tissue visualization has required that the tissue of interest be serially sectioned and imaged, subjecting each tissue section to unique non-linear deformations, dramatically hampering one's ability to evaluate cellular morphology, distribution and connectivity in the central nervous system (CNS). However, optical clearing techniques are changing the way tissues are visualized. These approaches permit one to probe deeply into intact organ preparations, providing tremendous insight into the structural organization of tissues in health and disease. Techniques such as Clear Lipid-exchanged Acrylamide-hybridized Rigid Imaging-compatible Tissue-hYdrogel (CLARITY) achieve this goal by providing a matrix that binds important biomolecules while permitting light-scattering lipids to freely diffuse out. Lipid removal, followed by refractive index matching, renders the tissue transparent and readily imaged in 3 dimensions (3D). Nevertheless, the electrophoretic tissue clearing (ETC) used in the original CLARITY protocol can be challenging to implement successfully and the use of a proprietary refraction index matching solution makes it expensive to use the technique routinely. This report demonstrates the implementation of a simple and inexpensive optical clearing protocol that combines passive CLARITY for improved tissue integrity and 2,2′-thiodiethanol (TDE), a previously described refractive index matching solution. Video LinkThe video component of this article can be found at

show abstract

Section: Discussionmentioning

confidence: 99%

Optical Clearing of the Mouse Central Nervous System Using Passive CLARITY

Roberts

Johnsonbaugh

Spence

et al. 2016

JoVE

View full text Add to dashboard Cite

show abstract

“…Most markers, including mTurquoise2, mNeonGreen, and mRuby2, can also be detected after mounting labeled tissues in optical clearing reagents like RIMS 24 ( Fig. 3b), ScaleSQ 25 (Figs.…”

Section: Evaluation Of Transgene Expressionmentioning

confidence: 99%

“…Commonly used reporters like GFP, eYFP, and tdTomato show strong fluorescent labeling in PACT-and PARS-cleared tissues, enabling whole-organ and thick-tissue imaging of transgene expression 3,22,24 . Most markers, including mTurquoise2, mNeonGreen, and mRuby2, can also be detected after mounting labeled tissues in optical clearing reagents like RIMS 24 ( Fig.…”

Section: Evaluation Of Transgene Expressionmentioning

confidence: 99%

“…To visualize these reporters, we suggest using optical clearing methods like RIMS (refractive index matching solution) 24 or ScaleSQ 25 (Figs. 3a and c, and 4b-c), or commercially available mounting media like Prolong Diamond Antifade (Thermo Fisher Scientific, cat.…”

Section: (Figs 2-4)mentioning

confidence: 99%

“…Inset shows zoomed-in view of ganglia containing TH + cell bodies; tdTomato-f labels both thick nerve bundles and individual fibers. Whole-mount tissues were optically cleared using either ScaleSQ 25 (a (right), c, and d) or RIMS 24 (b) and imaged using confocal microscopy, unless stated otherwise; all confocal images are presented as maximum intensity projections. Refer to peer-reviewed) is the author/funder.…”

Section: Supplementarymentioning

confidence: 99%

See 2 more Smart Citations

Widespread and targeted gene expression by systemic AAV vectors: Production, purification, and administration

Challis

Kumar

Chan

et al. 2018

Preprint

Self Cite

View full text Add to dashboard Cite

We recently developed novel AAV capsids for efficient and noninvasive gene transfer across the central and peripheral nervous systems. In this protocol, we describe how to produce and systemically administer AAV-PHP viruses to label and/or genetically manipulate cells in the mouse nervous system and organs including the heart. The procedure comprises three separate stages: AAV production, intravenous delivery, and evaluation of transgene expression.The protocol spans eight days, excluding the time required to assess gene expression, and can be readily adopted by laboratories with standard molecular and cell culture capabilities. We provide guidelines for experimental design and choosing the capsid, cargo, and viral dose appropriate for the experimental aims. The procedures outlined here are adaptable to diverse biomedical applications, from anatomical and functional mapping to gene expression, silencing, and editing.

show abstract

iRGD Modified Chemo‐immunotherapeutic Nanoparticles for Enhanced Immunotherapy against Glioblastoma

Kuang

Song

Yin

et al. 2018

Adv Funct Materials

106

View full text Add to dashboard Cite

Glioblastoma is the most common primary brain tumor in adults and still remains incurable, due to the limited accumulation of drugs in the tumor area. Herein, iRGD‐modified nanoparticles, DOX@MSN‐SS‐iRGD&1MT, are developed for simultaneous delivery of chemotherapeutic agents (doxorubicin, DOX) and immune checkpoint inhibitor (1‐methyltryptophan, 1MT) into orthotopic glioma. The nanoparticles are comprised of mesoporous silica nanoparticles loaded with DOX, combined with Asp‐Glu‐Val‐Asp (DEVD) connected 1MT, and finally modified by iRGD. These nanoparticles show the capability of penetrating through blood brain barrier into the tumor area, and significantly improve accumulation of drugs in orthotopic brain tumors with minimal side effects. The nanoparticles also activate cytotoxic CD8+ T lymphocytes and inhibit CD4+ T cells in both GL261 cells cocultured with splenocytes in vitro and GL261‐luc orthotopic tumors in vivo. Moreover, the expression of antitumor cytokines IFNα/β, IFN‐γ, TNF, IL‐17, STING, and GrzB is upregulated while protumor proteins p‐STAT3 and IL‐10 are downregulated in the brain tumor area. This study demonstrates the advantages of chemo‐immunotherapeutic nanoparticles accumulated in the brain tumor area and their effectively inhibiting tumor proliferation, which establishes a delivery platform to promote antitumor immunity against glioblastoma.

show abstract

Whole-body tissue stabilization and selective extractions via tissue-hydrogel hybrids for high-resolution intact circuit mapping and phenotyping

Cited by 234 publications

References 204 publications

Optical Clearing of the Mouse Central Nervous System Using Passive CLARITY

Optical Clearing of the Mouse Central Nervous System Using Passive CLARITY

Widespread and targeted gene expression by systemic AAV vectors: Production, purification, and administration

iRGD Modified Chemo‐immunotherapeutic Nanoparticles for Enhanced Immunotherapy against Glioblastoma

Contact Info

Product

Resources

About