4.6 Article

The rapid-tome, a 3D-printed microtome, and an updated hand-sectioning method for high-quality plant sectioning

期刊

PLANT METHODS
卷 19, 期 1, 页码 -

出版社

BMC
DOI: 10.1186/s13007-023-00986-3

关键词

Plant; Sectioning; Microscopy; Plant anatomy; 3D-printing; Root; Stem; Cross section

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Microscopic analysis of plant anatomy requires high-quality sections for accurate measurements. The authors present a hand-sectioning method suitable for softer tissues and a novel microtome design called Rapid-Tome for sectioning both soft and tougher plant materials. The Rapid-Tome allows rapid sectioning, sample control, and is cost-effective compared to traditional microtomes. High school students were able to produce sections for comparative analyses with minimal training.
BackgroundMicroscopic analysis of plant anatomy is a common procedure in biology to study structure and function that requires high-quality sections for accurate measurements. Hand sectioning of specimens is typically limited to moderately soft tissue while harder samples prohibit sectioning by hand and/or result in inconsistent thicknesses.ResultsHere we present both a clearly described hand-sectioning method and a novel microtome design that together provide the means to section a variety of plant sample types. The described hand-sectioning method for herbaceous stems works well for softer subjects but is less suitable for samples with secondary growth (e.g., wood production). Instead, the Rapid-Tome is a novel tool for sectioning both soft and tougher high-aspect-ratio samples, such as stems and roots, with excellent sample control. The Rapid-Tome can be 3D-printed in approximately 18 h on a mid-quality printer common at university maker spaces. After printing and trimming, Rapid-Tome assembly takes a few minutes with five metal parts common at hardware stores. Users sectioned a variety of plant samples including the hollow internodes of switchgrass (Panicum virgatum), fibrous switchgrass roots containing aerenchyma, and woody branches of eastern red cedar (Juniperus virginiana) and American sycamore (Platanus occidentalis). A comparative analyses with Rapid-Tome-produced sections readily revealed a significant difference in seasonal growth of sycamore xylem vessel area in spring (49%) vs. summer (23%). Additionally, high school students with no prior experience produced sections with the Rapid-Tome adequate for comparative analyses of various plant samples in less than an hour.ConclusionsThe described hand-sectioning method is suitable for softer tissues, including hollow-stemmed grasses and similar samples. In addition, the Rapid-Tome provides capacity to safely produce high-quality sections of tougher plant materials at a fraction of the cost of traditional microtomes combined with excellent sample control. The Rapid-Tome features rapid sectioning, sample advancement, blade changes, and sample changes; it is highly portable and can be used easily with minimal training making production of thin sections accessible for classroom and outreach use, in addition to research.

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