Abstract
Ubiquitous in eukaryotic organisms, the flagellum is a well-studied organelle that is well-known to be responsible for motility in a variety of organisms. Commonly necessitated in their study is the capability to image and subsequently track the movement of one or more flagella using videomicroscopy, requiring digital isolation and location of the flagellum within a sequence of frames. Such a process in general currently requires some researcher input, providing some manual estimate or reliance on an experiment-specific heuristic to correctly identify and track the motion of a flagellum. Here we present a fully-automated method of flagellum identification from videomicroscopy based on the fact that the flagella are of approximately constant width when viewed by microscopy. We demonstrate the effectiveness of the algorithm by application to captured videomicroscopy of Leishmania mexicana, a parasitic monoflagellate of the family Trypanosomatidae. ImageJ Macros for flagellar identification are provided, and high accuracy and remarkable throughput are achieved via this unsupervised method, obtaining results comparable in quality to previous studies of closely-related species but achieved without the need for precursory measurements or the development of a specialised heuristic, enabling in general the automated generation of digitised kinematic descriptions of flagellar beating from videomicroscopy.
Highlights
Well-studied and present across a wide range of organisms, the eukaryotic flagellum is typically a long slender organelle that is known to perform a variety of functional roles throughout nature, perhaps most notably in spermatozoa where the beating of one or more flagella can render a gamete motile[1]
Sample macros may be found in the Supplementary Information, where image preprocessing and flagellum tracing have been implemented in a basic manner that is sufficient for exemplifying the flagellar identification method
In this work we have presented and verified a method of flagellar identification from the videomicroscopy of free-swimming bodies
Summary
Well-studied and present across a wide range of organisms, the eukaryotic flagellum is typically a long slender organelle that is known to perform a variety of functional roles throughout nature, perhaps most notably in spermatozoa where the beating of one or more flagella can render a gamete motile[1]. Image transforms, such as the Hough and Radon transforms, are commonly used for feature extraction and image processing. Owing to the noted characteristic shape of flagella, there is significant potential utility in this shape-encoding image transform in the context of flagellum tracking and identification
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