Studying neuronal processes such as synaptic summation, dendritic physiology and neural network dynamics requires complex spatiotemporal control over neuronal activities. The recent development of neural photosensitization tools, such as channelrhodopsin-2 (ChR2), offers new opportunities for non-invasive, flexible and cell-specific neuronal stimulation. Previously, complex spatiotemporal control of photosensitized neurons has been limited by the lack of appropriate optical devices which can provide 2D stimulation with sufficient irradiance. Here we present a simple and powerful solution that is based on an array of high-power micro light-emitting diodes (micro-LEDs) that can generate arbitrary optical excitation patterns on a neuronal sample with micrometre and millisecond resolution. We first describe the design and fabrication of the system and characterize its capabilities. We then demonstrate its capacity to elicit precise electrophysiological responses in cultured and slice neurons expressing ChR2.
Stimulating neuron cells with light is an exciting new technology that is revolutionizing the neurosciences. To date, due to the optical complexity that is involved, photostimulation has only been achieved at a single site using high power light sources. Here we present a GaN based micro-light emitting diode (LED) array that can open the way to multi-site photostimulation of neuron cells. The device is a two-dimensional array of micrometre size LED emitters. Each emitter has the required wavelength, optical power and modulation bandwidth to trigger almost any photosensitizer and is individually addressable. We demonstrate micrometre resolution photoactivation of a caged fluorophore and photostimulation of sensitized living neuron cells. In addition, a complete system that combines the micro-LED array with multi-site electrophysiological recording based on microelectrode array technology and/or fluorescence imaging is presented.
Lensless on-chip imaging is a promising technique to count and monitor cells and micro-objects in liquid sample. In this paper we apply this technique to the observation of µL sample containing bacteria evaporated onto a microscope slide. Compared with previously reported techniques, a large improvement in signal to noise ratio is obtained due to the presence of a few μm thick wetting film creating a micro-lens on top of each bacteria. In these conditions, standard CMOS sensor are able to detect micro-objects as small as few μm, e.g. E.coli and Bacillus subtilis bacteria and 1 μm polymer beads with a large signal to noise ratio of 45 ± 10. An overall detection efficiency of 85 ± 7% and a co-localization error of σ1D = 1.1μm compared with reference fluorescence microscopy images are achieved. This novel technique will be used as a pre-positioning tool prior to other optical identification methods, e.g. Raman spectroscopy.
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