Development of single-beam wide-field infrared imaging to study sub-cellular neuron biochemistry

Multi-beam wide-field imaging using synchrotron mid-infrared light sources coupled with focal plane array detectors has provided a major breakthrough to the field of bio-spectroscopic imaging. The ability to collect sub-cellular molecular images in minutes has opened the door to a new era of biochemical studies. Although a multi-beam approach is the superior method to this form of imaging, it requires a specialized set of beamline optics, which may not be compatible with existing mid-infrared microscopy beamlines, or research programs/applications currently in place (some of which do not require an imaging component). In this investigation we demonstrate that a single-beam approach can be utilized in a similar manner to multi-beam imaging, to collect sub-cellular biochemical images of brain neurons in a rapid time frame, without extensive modification of an existing beamline configuration. This study uses an applied example, imaging the same neuron in situ within a brain tissue section, with both synchrotron and thermal sources. The results highlight the advantage of improved spatial resolution/image quality and spectral quality (signal to noise ratio) that is obtained when a high magnification and high numerical aperture objective (52×, 0.65) is coupled to a synchrotron mid-infrared lightsource with a focal plane array detector. The approach we report may prove to be particularly appealing to numerous existing mid-infrared beamlines, allowing straightforward integration of sub-cellular biochemical imaging with existing non-imaging research applications.