PEA-CLARITY: 3D molecular imaging of whole plant organs.
Here we report the adaptation of the CLARITY technique to plant tissues with addition of enzymatic degradation to improve optical clearing and facilitate antibody probe penetration. Plant-Enzyme-Assisted (PEA)-CLARITY, has allowed deep optical visualisation of stains, expressed fluorescent proteins and IgG-antibodies in Tobacco and Arabidopsis leaves. Enzyme treatment enabled penetration of antibodies into whole tissues without the need for any sectioning of the material, thus facilitating protein localisation of intact tissue in 3D whilst retaining cellular structure.
School of Environmental and Life Sciences, University of Newcastle, Callaghan, NSW, 2308, Australia.
William M
WM
Palmer
School of Environmental and Life Sciences, University of Newcastle, Callaghan, NSW, 2308, Australia.
Antony P
AP
Martin
School of Biomedical Sciences and Pharmacy, University of Newcastle, Callaghan, NSW 2308, Australia.
Jamie R
JR
Flynn
School of Environmental and Life Sciences, University of Newcastle, Callaghan, NSW, 2308, Australia.
Stephanie L
SL
Reed
CSIRO Agriculture, Black Mountain, ACT, 2601, Australia.
Rosemary G
RG
White
ARC Centre of Excellence for Translational Photosynthesis, Australian National University, Acton, ACT, 2601, Australia.
Robert T
RT
Furbank
School of Environmental and Life Sciences, University of Newcastle, Callaghan, NSW, 2308, Australia.
Christopher P L
CPL
Grof
D017360:Arabidopsis / Q000033:anatomy & histology*; D049452:Green Fluorescent Proteins / Q000378:metabolism; D021621:Imaging, Three-Dimensional; D057054:Molecular Imaging; D061848:Optical Imaging; D009928:Organ Specificity; D012984:Software; D014026:Nicotiana / Q000033:anatomy & histology*
D049452:Green Fluorescent Proteins