Clear heads: simplified tissue preparation for studying plant anatomy
and development in the Asteraceae

Reid Selby

Department of Biological Sciences, Clemson University, Clemson SC, USA
Department of Biological Sciences, Auburn University, Auburn AL, USA

https://orcid.org/0009-0005-3690-8955

Brannan R. Cliver

Department of Biological Sciences, Clemson University, Clemson SC, USA
Department of Biological Sciences, Auburn University, Auburn AL, USA

https://orcid.org/0009-0002-4183-2789

Zach Meharg

Department of Crop, Soil, and Environmental Science, Auburn University, Auburn AL, USA
HudsonAlpha Institute for Biotechnology, Huntsville AL, USA

https://orcid.org/0009-0004-7382-8527

Alex Harkess

HudsonAlpha Institute for Biotechnology, Huntsville AL, USA

https://orcid.org/0000-0002-2035-0871

Daniel S. Jones

Department of Biological Sciences, Clemson University, Clemson SC, USA
Department of Biological Sciences, Auburn University, Auburn AL, USA

https://orcid.org/0000-0002-9241-1813

DOI: http://dx.doi.org/10.53875/capitulum.04.1.02

ABSTRACT:  The use of microscopy is instrumental for our understanding of plant anatomy and development. A major bottleneck is that conventional methods for plant tissue preparation can take weeks to complete and can involve hazardous materials that many labs are not equipped to handle. Furthermore, these methods traditionally involve embedding the fixed tissue in embedding substrates such as paraffin or plastic resins, which is both a laborious and time-intensive process. Here we describe a simplified and accessible method that accomplishes tissue fixation and clearing in just three days, minimizes the use of hazardous materials, and eliminates the need for embedding/sectioning. This method also utilizes structural autofluorescence of plant tissues and thus does not require any counterstaining to resolve structure. We demonstrate the utility of this preparation using both epifluorescence and confocal microscopy on examples from the Asteraceae family, making both techniques more approachable to biologists of all backgrounds and thus enabling a richer exploration of plant anatomy and development.

Keywords:  anatomy, confocal, development, fluorescence microscopy, morphological diversity, tissue clearing.