A versatile vessel casting method for fine mapping of vascular networks using a hydrogel-based lipophilic dye solution

Zhu, Jingtan; Liu, Xiaomei; Xu, Jianyi; Deng, Yating; Wang, Pingfu; Liu, Zhang; Yang, Qing; Li, Dongyu; Yu, Tingting; Zhu, Dan

doi:10.1016/j.crmeth.2023.100407

Cited by 8 publications

(5 citation statements)

References 57 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…In our previous publications, 1 we have demonstrated that VALID successfully restrained elution against organic solvents and were finely retained without apparent loss after clearing, therefore it is suitable for different tissue clearing protocols. Combining VALID with tissue clearing and light sheet microscopy allows 3D imaging and analysis of different vascular networks under physiological and pathological state.…”

Section: Expected Outcomesmentioning

confidence: 94%

Protocol for fine casting, imaging, and analysis of murine vascular networks with VALID

Zhu

Liu

et al. 2023

STAR Protocols

Self Cite

View full text Add to dashboard Cite

Section: Expected Outcomesmentioning

confidence: 94%

Protocol for fine casting, imaging, and analysis of murine vascular networks with VALID

Zhu

Liu

et al. 2023

STAR Protocols

Self Cite

View full text Add to dashboard Cite

“…Three days later, mice underwent deep sedation via intraperitoneal administration of sodium pentobarbital at a dosage of 50 mg/kg. Following this, vascular labeling was executed utilizing the VALID protocol [14]. Initially, mice were infused with 0.01 M phosphate-buffered saline (PBS) for the removal of blood from the vascular system.…”

Section: Vascular Labeling and Tissue Clearingmentioning

confidence: 99%

A novel method (RIM-Deep) for enhancing imaging depth and resolution stability of deep cleared tissue in inverted confocal microscopy

Liu,

Wang,

Zou

et al. 2024

Preprint

View full text Add to dashboard Cite

The increasing use of tissue clearing techniques underscores the urgent need for cost-effective and simplified deep imaging methods. While traditional inverted confocal microscopes excel in high-resolution imaging of tissue sections and cultured cells, they face limitations in deep imaging of cleared tissues due to refractive index mismatches between the immersion media of objectives and sample container. To overcome these challenges, the RIM-Deep was developed to significantly improve deep imaging capabilities without compromising the normal function of the confocal microscope. This system facilitates deep immunofluorescence imaging of the prefrontal cortex in cleared macaque tissue, extending imaging depth from 2 mm to 5 mm. Applied to an intact and cleared Thy1-EGFP mouse brain, the system allowed for clear axonal visualization at high imaging depth. Moreover, this advancement enables large-scale, deep 3D imaging of intact tissues. In principle, this concept can be extended to any imaging modality, including existing inverted wide-field, confocal, and two-photon microscopy. This would significantly upgrade traditional laboratory configurations and facilitate the study of connectomics in the brain and other tissues.

show abstract

“…Perturbations in the vascular network’s architecture are intricately linked to the onset and progression of diverse ailments such as cancer metastasis, stroke, diabetes, Alzheimer’s disease, etc. Hence, comprehensive visualization of the whole-body vascular network at the capillary scale is imperative for elucidating the underlying mechanisms of various vascular-related disorders and devising precise therapeutic interventions. − …”

Section: Introductionmentioning

confidence: 99%

Light-Sheet Microscopic Imaging of Whole-Mouse Vascular Network with Fluorescent Microsphere Perfusion

Cao,

Li,

et al. 2024

ACS Biomater. Sci. Eng.

View full text Add to dashboard Cite

Visualizing the whole vascular network system is crucial for understanding the pathogenesis of specific diseases and devising targeted therapeutic interventions. Although the combination of light sheet microscopy and tissue-clearing methods has emerged as a promising approach for investigating the blood vascular network, leveraging the spatial resolution down to the capillary level and the ability to image centimeter-scale samples remains difficult. Especially, as the resolution improves, the issue of photobleaching outside the field of view poses a challenge to image the whole vascular network of adult mice at capillary resolution. Here, we devise a fluorescent microsphere vascular perfusion method to enable labeling of the whole vascular network in adult mice, which overcomes the photobleaching limit during the imaging of large samples. Moreover, by combining the utilization of a large-scale light-sheet microscope and tissue clearing protocols for whole-mouse samples, we achieve the capillary-level imaging resolution (3.2 × 3.2 × 6.5 μm) of the whole vascular network with dimensions of 45 × 15 × 82 mm in adult mice. This method thus holds great potential to deliver mesoscopic resolution images of various tissue organs for whole-animal imaging.

show abstract

A versatile vessel casting method for fine mapping of vascular networks using a hydrogel-based lipophilic dye solution

Cited by 8 publications

References 57 publications

Protocol for fine casting, imaging, and analysis of murine vascular networks with VALID

Protocol for fine casting, imaging, and analysis of murine vascular networks with VALID

A novel method (RIM-Deep) for enhancing imaging depth and resolution stability of deep cleared tissue in inverted confocal microscopy

Light-Sheet Microscopic Imaging of Whole-Mouse Vascular Network with Fluorescent Microsphere Perfusion

Contact Info

Product

Resources

About