An updated protocol for high throughput plant tissue sectioning
Quantification of the tissue and cellular structure of plant material is essential for the study of a variety of plant sciences applications. Currently, many methods for sectioning plant material are either low throughput or involve free-hand sectioning which requires a significant amount of practic...
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Frontiers Media
2017
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| Online Access: | https://eprints.nottingham.ac.uk/47000/ |
| _version_ | 1848797446355812352 |
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| author | Atkinson, Jonathan A. Wells, Darren M. |
| author_facet | Atkinson, Jonathan A. Wells, Darren M. |
| author_sort | Atkinson, Jonathan A. |
| building | Nottingham Research Data Repository |
| collection | Online Access |
| description | Quantification of the tissue and cellular structure of plant material is essential for the study of a variety of plant sciences applications. Currently, many methods for sectioning plant material are either low throughput or involve free-hand sectioning which requires a significant amount of practice. Here, we present an updated method to provide rapid and high-quality cross sections, primarily of root tissue but which can also be readily applied to other tissues such as leaves or stems. To increase the throughput of traditional agarose embedding and sectioning, custom designed 3D printed molds were utilized to embed 5–15 roots in a block for sectioning in a single cut. A single fluorescent stain in combination with laser scanning confocal microscopy was used to obtain high quality images of thick sections. The provided CAD files allow production of the embedding molds described here from a number of online 3D printing services. Although originally developed for roots, this method provides rapid, high quality cross sections of many plant tissue types, making it suitable for use in forward genetic screens for differences in specific cell structures or developmental changes. To demonstrate the utility of the technique, the two parent lines of the wheat (Triticum aestivum) Chinese Spring × Paragon doubled haploid mapping population were phenotyped for root anatomical differences. Significant differences in adventitious cross section area, stele area, xylem, phloem, metaxylem, and cortical cell file count were found. |
| first_indexed | 2025-11-14T20:04:00Z |
| format | Article |
| id | nottingham-47000 |
| institution | University of Nottingham Malaysia Campus |
| institution_category | Local University |
| last_indexed | 2025-11-14T20:04:00Z |
| publishDate | 2017 |
| publisher | Frontiers Media |
| recordtype | eprints |
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| spelling | nottingham-470002020-05-04T19:10:47Z https://eprints.nottingham.ac.uk/47000/ An updated protocol for high throughput plant tissue sectioning Atkinson, Jonathan A. Wells, Darren M. Quantification of the tissue and cellular structure of plant material is essential for the study of a variety of plant sciences applications. Currently, many methods for sectioning plant material are either low throughput or involve free-hand sectioning which requires a significant amount of practice. Here, we present an updated method to provide rapid and high-quality cross sections, primarily of root tissue but which can also be readily applied to other tissues such as leaves or stems. To increase the throughput of traditional agarose embedding and sectioning, custom designed 3D printed molds were utilized to embed 5–15 roots in a block for sectioning in a single cut. A single fluorescent stain in combination with laser scanning confocal microscopy was used to obtain high quality images of thick sections. The provided CAD files allow production of the embedding molds described here from a number of online 3D printing services. Although originally developed for roots, this method provides rapid, high quality cross sections of many plant tissue types, making it suitable for use in forward genetic screens for differences in specific cell structures or developmental changes. To demonstrate the utility of the technique, the two parent lines of the wheat (Triticum aestivum) Chinese Spring × Paragon doubled haploid mapping population were phenotyped for root anatomical differences. Significant differences in adventitious cross section area, stele area, xylem, phloem, metaxylem, and cortical cell file count were found. Frontiers Media 2017-10-04 Article PeerReviewed Atkinson, Jonathan A. and Wells, Darren M. (2017) An updated protocol for high throughput plant tissue sectioning. Frontiers in Plant Science, 8 . p. 1721. ISSN 1664-462X Tissue sectioning root anatomy cross section confocal microscopy 3D printing https://www.frontiersin.org/articles/10.3389/fpls.2017.01721/full doi:10.3389/fpls.2017.01721 doi:10.3389/fpls.2017.01721 |
| spellingShingle | Tissue sectioning root anatomy cross section confocal microscopy 3D printing Atkinson, Jonathan A. Wells, Darren M. An updated protocol for high throughput plant tissue sectioning |
| title | An updated protocol for high throughput plant tissue sectioning |
| title_full | An updated protocol for high throughput plant tissue sectioning |
| title_fullStr | An updated protocol for high throughput plant tissue sectioning |
| title_full_unstemmed | An updated protocol for high throughput plant tissue sectioning |
| title_short | An updated protocol for high throughput plant tissue sectioning |
| title_sort | updated protocol for high throughput plant tissue sectioning |
| topic | Tissue sectioning root anatomy cross section confocal microscopy 3D printing |
| url | https://eprints.nottingham.ac.uk/47000/ https://eprints.nottingham.ac.uk/47000/ https://eprints.nottingham.ac.uk/47000/ |