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A rapid, nondestructive method for vascular network visualization.

ABSTRACT: The quantitative analysis of blood vessel networks is an important component in many animal models of disease. We describe a nondestructive technique for blood vessel imaging that visualizes in situ vasculature in harvested tissues. The method allows for further analysis of the same tissues with histology and other methods that can be performed on fixed tissue. Consequently, it can easily be incorporated upstream to analysis methods to augment these with a three-dimensional reconstruction of the vascular network in the tissues to be analyzed. The method combines iodine-enhanced micro-computed tomography with a deep learning algorithm to segment vasculature within tissues. The procedure is relatively simple and can provide insight into complex changes in the vascular structure in the tissues.

SUBMITTER: Veith A

PROVIDER: S-EPMC7807294 | biostudies-literature | 2020 Dec

REPOSITORIES: biostudies-literature

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A rapid, nondestructive method for vascular network visualization.

Veith Austin A B Baker Aaron A

BioTechniques 20201028 6

The quantitative analysis of blood vessel networks is an important component in many animal models of disease. We describe a nondestructive technique for blood vessel imaging that visualizes <i>in situ</i> vasculature in harvested tissues. The method allows for further analysis of the same tissues with histology and other methods that can be performed on fixed tissue. Consequently, it can easily be incorporated upstream to analysis methods to augment these with a three-dimensional reconstruction ...[more]

PMID: 33108897

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Project description:The agar culture plate has played a crucial role in bacteriology since the origins of the discipline and is a staple bioanalytical method for efforts ranging from research to standard clinical diagnostic tests. However, plating, inoculating, and waiting for microbes to develop colonies that are visible is time-consuming. In this work, we demonstrate white-light interferometry (WLI) as a practical tool for accelerated and improved measurement of bacterial cultures. High resolution WLI surface profile imaging was used for nondestructive characterization and counting of bacterial colonies on agar before they became visible to the naked eye. The three-dimensional (3D) morphology of Gram-negative (Pseudomonas fluorescens) and Gram-positive (Bacillus thuringiensis) bacterial species were monitored with WLI over time by collecting surface profiles of colonies on agar plates with high vertical resolution (3-5 nanometers) and large field of view (3-5 mm). This unique combination of sensitive vertical resolution and large field of view uniquely provided by WLI enables measurement of colony morphologies and nondestructive monitoring of hundreds of microcolonies. Individual bacteria were imaged within the first few hours after plating and colonies were accurately counted with results comparing favorably to counts made by traditional methods that require much longer wait times. Nondestructive imaging was used to track single cells multiplying into small colonies and the volume changes over time in these colonies were used to measure their growth rates. Based on the results herein, bioimaging with WLI was demonstrated as a novel rapid bacterial culture assay with several advantageous capabilities. Fast nondestructive counting of colony-forming units in a culture and simultaneous measurement of bacterial growth rates and colony morphology with this method may be beneficial in research and clinical applications where current methods are either too slow or are destructive.

Dataset Information

A rapid, nondestructive method for vascular network visualization.

Publications

A rapid, nondestructive method for vascular network visualization.

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