Abstract
Abstract
Background
Microscopic 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.
Results
Here 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.
Conclusions
The 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.
Funder
U.S. Department of Energy
National Institute of Food and Agriculture
Publisher
Springer Science and Business Media LLC
Subject
Plant Science,Genetics,Biotechnology
Reference25 articles.
1. Huang C, Wang Z, Quinn D, Suresh S, Hsia KJ. Differential growth and shape formation in plant organs. Proc Natl Acad Sci. 2018;115(49):12359–64.
2. Teague BP, Guye P, Weiss R. Synthetic morphogenesis. Cold Spring Harb Perspect Biol. 2016;8(9): a023929.
3. Schneeberger K. Using next-generation sequencing to isolate mutant genes from forward genetic screens. Nat Rev Genet. 2014;15(10):662–76.
4. Turner SR, Somerville CR. Collapsed xylem phenotype of Arabidopsis identifies mutants deficient in cellulose deposition in the secondary cell wall. Plant Cell. 1997;9(5):689–701.
5. Sakai K, Citerne S, Antelme S, Le Bris P, Daniel S, Bouder A, D’Orlando A, Cartwright A, Tellier F, Pateyron S, et al. BdERECTA controls vasculature patterning and phloem-xylem organization in Brachypodium distachyon. BMC Plant Biol. 2021;21(1):196.