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Synchrotron radiation-based infrared microspectroscopy (SR-IMS) is a nondestructive bioanalytical technique with a high signal-to-noise ratio and high ultraspatial resolution (3-10 μm). It is capable to explore the microstructures of plant tissues in a chemical sense and provide information on the composition, structure, and distribution of chemical compounds/functional groups. The objective of this study was to illustrate how SR-IMS can be used to image the internal microstructures of chickpea seed tissue within a cellular level. Chickpea seeds (CDC Cory) were collected from the Crop Development Center (University of Saskatchewan, Saskatoon, SK). The seeds were frozen at -20 °C on object disks in a cryostatic microtome and then were cut into thin cross sections ( ca . 8 μm thick). The experiment was carried out on the mid-infrared beamline (01B1-1) at the Canadian Light Source (Saskatoon, SK). We obtained the ultraspatial images of the chickpea tissue with pixel-sized increments of imaging steps. The results showed that, with the extremely bright synchrotron light, spectra with high signal-to-noise ratios can be obtained from an area as small as 3.3 μm × 3.3 μm, allowing us to observe the seed tissue within a cellular level. Chemical distribution of chickpea such as lipids, protein, and carbohydrates could be mapped, revealing the chemical information of the chickpea internal microstructure. In conclusion, SR-IMS can rapidly characterize the molecular structure of protein, carbohydrates, and lipids at an ultraspatial resolution.

journal of agricultural and food chemistry

Chemical Imaging of the Microstructure of Chickpea Seed Tissue within a Cellular Dimension Using Synchrotron Infrared Microspectroscopy: A Preliminary Study