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Abstract Alterations in vascular networks, including angiogenesis and capillary regression, play key roles in disease, wound healing, and development. The spatial structures of blood vessels can be captured through imaging, but effective characterization of network architecture requires both metrics for quantification and software to carry out the analysis in a high‐throughput and unbiased fashion. We present Rapid Editable Analysis of Vessel Elements Routine (REAVER), an open‐source tool that researchers can use to analyze high‐resolution 2D fluorescent images of blood vessel networks, and assess its performance compared to alternative image analysis programs. Using a dataset of manually analyzed images from a variety of murine tissues as a ground‐truth, REAVER exhibited high accuracy and precision for all vessel architecture metrics quantified, including vessel length density, vessel area fraction, mean vessel diameter, and branchpoint count, along with the highest pixel‐by‐pixel accuracy for the segmentation of the blood vessel network. In instances where REAVER's automated segmentation is inaccurate, we show that combining manual curation with automated analysis improves the accuracy of vessel architecture metrics. REAVER can be used to quantify differences in blood vessel architectures, making it useful in experiments designed to evaluate the effects of different external perturbations (eg, drugs or disease states).
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This content will become publicly available on January 1, 2026
Quantification of variegated Drosophila ommatidia with high-resolution image analysis and machine learning
A longstanding challenge in biology is accurately analyzing images acquired using microscopy. Recently, machine learning (ML) approaches have facilitated detailed quantification of images that were refractile to traditional computation methods. Here, we detail a method for measuring pigments in the complex-mosaic adult Drosophila eye using high-resolution photographs and the pixel classifier ilastik [1]. We compare our results to analyses focused on pigment biochemistry and subjective interpretation, demonstrating general overlap, while highlighting the inverse relationship between accuracy and high-throughput capability of each approach. Notably, no coding experience is necessary for image analysis and pigment quantification. When considering time, resolution, and accuracy, our view is that ML-based image analysis is the preferred method.
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- Award ID(s):
- 2145195
- PAR ID:
- 10583196
- Publisher / Repository:
- Oxford Academic
- Date Published:
- Journal Name:
- Biology Methods and Protocols
- Volume:
- 10
- Issue:
- 1
- ISSN:
- 2396-8923
- Format(s):
- Medium: X
- Sponsoring Org:
- National Science Foundation
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