Optical dissection of stimulus-evoked retinal activation

Department of Biomedical Engineering, University of Alabama at Birmingham, Birmingham, AL 35294, USA.
Optics Express (Impact Factor: 3.49). 09/2008; 16(17):12446-59. DOI: 10.1364/OE.16.012446
Source: PubMed


Better understanding of stimulus-evoked intrinsic optical signals (IOSs) in the retina promises new methodology for study and diagnosis of retinal function. Using a flood-illumination near infrared (NIR) light microscope equipped with high-speed CCD (80 Hz) and CMOS (1000 Hz) cameras, we validated depth-resolved enface imaging of fast IOSs in isolated retina of leopard frog. Both positive (increasing) and negative (decreasing) IOSs were observed at the photoreceptor and inner layers of the retina. The distribution of IOSs with opposite polarities showed a center-surround pattern. At the photoreceptor layer, negative IOSs dominated the center area illuminated by the stimulus light spot, while positive signals dominated the surrounding area. In contrast, at inner retinal layers, positive IOSs dominated the center area covered by the stimulus light spot, and negative IOSs were mainly observed in the surrounding area. Fast CMOS imaging disclosed rapid IOSs within 5 ms after the stimulus onset, and both ON and OFF optical responses were observed associated with a step light stimulus.

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    • "The experimental procedures were approved by the Institutional Animal Care and Use Committee of University of Alabama at Birmingham. Details of the preparation of flat-mounted retinas [34] and retinal slices [31] have been previously reported. Briefly, retinal dissection was conducted in a dark room with dim red illumination. "
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    ABSTRACT: The purpose of this study was to investigate cellular sources of autofluorescence signals in freshly isolated frog (Rana pipiens) retinas. Equipped with an ultrafast laser, a laser scanning two-photon excitation fluorescence microscope was employed for sub-cellular resolution examination of both sliced and flat-mounted retinas. Two-photon imaging of retinal slices revealed autofluorescence signals over multiple functional layers, including the photoreceptor layer (PRL), outer nuclear layer (ONL), outer plexiform layer (OPL), inner nuclear layer (INL), inner plexiform layer (IPL), and ganglion cell layer (GCL). Using flat-mounted retinas, depth-resolved imaging of individual retinal layers further confirmed multiple sources of autofluorescence signals. Cellular structures were clearly observed at the PRL, ONL, INL, and GCL. At the PRL, the autofluorescence was dominantly recorded from the intracellular compartment of the photoreceptors; while mixed intracellular and extracellular autofluorescence signals were observed at the ONL, INL, and GCL. High resolution autofluorescence imaging clearly revealed mosaic organization of rod and cone photoreceptors; and sub-cellular bright autofluorescence spots, which might relate to connecting cilium, was observed in the cone photoreceptors only. Moreover, single-cone and double-cone outer segments could be directly differentiated.
    Biomedical Optics Express 06/2011; 2(6):1494-503. DOI:10.1364/BOE.2.001494 · 3.65 Impact Factor
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    • "IOS imaging has been established for high resolution detection of stimulus-evoked physiological responses in the retina [13, 14] and other neural tissues [15–17]. Fast IOSs have time courses that are comparable to stimulus-evoked electrophysiological kinetics [18, 19], and allows simultaneous monitoring of coherent interactions of multiple neurons working together [20]. Given the fact that neurons and islet β-cells share many biochemical and molecular mechanisms, including their electrophysiological oscillation activities, we hypothesized that IOS imaging could provide a new method for high resolution evaluation of glucose-evoked responses in the islet β-cells. "
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    ABSTRACT: Simultaneous monitoring of many functioning β-cells is essential for understanding β-cell dysfunction as an early event in the progression to diabetes. Intrinsic optical signal (IOS) imaging has been shown to allow high resolution detection of stimulus-evoked physiological responses in the retina and other neural tissues. In this paper, we demonstrate the feasibility of using IOS imaging for functional examination of insulin secreting INS-1 cells, a popular model for investigating diabetes associated β-cell dysfunction. Our experiments indicate that IOS imaging permits simultaneous monitoring of glucose-stimulated physiological responses in multiple cells with high spatial (sub-cellular) and temporal (sub-second) resolution. Rapid IOS image sequences revealed transient optical responses that had time courses tightly correlated with the glucose stimulation.
    Optics Express 01/2011; 19(1):99-106. DOI:10.1364/OE.19.000099 · 3.49 Impact Factor
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    • "are stimulus-evoked optical responses represented in the unit of ΔI/I, where ΔI is the dynamic optical changes and I is the background light intensity. The IOS images were constructed using the following procedure [27]: 1) The images from the 0.5-s pre-stimulus baseline recording phase were averaged, pixel by pixel, and the averaged intensity of each pixel was taken as the background intensity I of each pixel; 2) The background intensity I was subtracted from each subsequent recorded frame, pixel by pixel, to get the ΔI of each pixel. 3) The ΔI/I image sequence was constructed to show the dynamic IOS patterns of the retina. "
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    ABSTRACT: High resolution monitoring of stimulus-evoked retinal neural activities is important for understanding retinal neural mechanisms, and can be a powerful tool for retinal disease diagnosis and treatment outcome evaluation. Fast intrinsic optical signals (IOSs), which have the time courses comparable to that of electrophysiological activities in the retina, hold the promise for high resolution imaging of retinal neural activities. However, application of fast IOS imaging has been hindered by the contamination of slow, high magnitude optical responses associated with transient hemodynamic and metabolic changes. In this paper we demonstrate the feasibility of separating fast retinal IOSs from slow optical responses by combining flicker stimulation and dynamic (temporal) differential image processing. A near infrared flood-illumination microscope equipped with a high-speed (1000 Hz) digital camera was used to conduct concurrent optical imaging and ERG measurement of isolated frog retinas. High spatiotemporal resolution imaging revealed that fast IOSs could follow flicker frequency up to at least 6 Hz. Comparable time courses of fast IOSs and ERG kinetics provide evidence that fast IOSs are originated from stimulus activated retinal neurons.
    Optics Express 03/2010; 18(7):7210-8. DOI:10.1364/OE.18.007210 · 3.49 Impact Factor
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