Abstract
Plants contain abundant autofluorescent molecules that can be used for biochemical, physiological, or imaging studies. The two most studied molecules are chlorophyll (orange/red fluorescence) and lignin (blue/green fluorescence). Chlorophyll fluorescence is used to measure the physiological state of plants using handheld devices that can measure photosynthesis, linear electron flux, and CO2 assimilation by directly scanning leaves, or by using reconnaissance imaging from a drone, an aircraft or a satellite. Lignin fluorescence can be used in imaging studies of wood for phenotyping of genetic variants in order to evaluate reaction wood formation, assess chemical modification of wood, and study fundamental cell wall properties using Förster Resonant Energy Transfer (FRET) and other methods. Many other fluorescent molecules have been characterized both within the protoplast and as components of cell walls. Such molecules have fluorescence emissions across the visible spectrum and can potentially be differentiated by spectral imaging or by evaluating their response to change in pH (ferulates) or chemicals such as Naturstoff reagent (flavonoids). Induced autofluorescence using glutaraldehyde fixation has been used to enable imaging of proteins/organelles in the cell protoplast and to allow fluorescence imaging of fungal mycelium.
Highlights
Autofluorescent molecules are common in plant tissues [1]
The two most important autofluorescent molecules found in plants are chlorophyll and lignin but a wide range of other molecules are autofluorescent with UV or visible excitation including components of both cytoplasm and cell walls (Figure 1) [1]
Autofluorescence should be examined on fresh tissue as some autofluorescent compounds can become redistributed or be completely removed from tissue when exposed to solvent-based fixatives such as formalin aceto-alcohol (FAA) or mounting media such as glycerol—instead, aqueous buffers can be used as mounting media [14,15]
Summary
Autofluorescent molecules are common in plant tissues [1]. This can be viewed from two opposing perspectives. Autofluorescence can be used as a label-free method for detecting specific molecules using spectral fluorescence imaging techniques and can be considered advantageous for some experimental investigations [1,5,6,7,8,9,10,11,12]. Autofluorescence should be examined on fresh tissue as some autofluorescent compounds (chlorophyll, flavonoids) can become redistributed or be completely removed from tissue when exposed to solvent-based fixatives such as formalin aceto-alcohol (FAA) or mounting media such as glycerol—instead, aqueous buffers can be used as mounting media [14,15]. 2 of aceto when exposed to solvent-based fixatives such as formalin alcohol (FAA) or mounting media such as glycerol—instead, aqueous buffers can be used as completely removed from tissue when exposed to solvent-based fixatives such as formalin acetomounting media [14,15]. 2.2.Chlorophyll and Chlorophyll autofluorescence usedboth bothininlab-scale lab-scaleexperiments experimentsand andininremote remotesensing sensing
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