Article

Fluorescence/Luminescence Circadian Imaging of Complex Tissues at Single-Cell Resolution

Journal of Biological Rhythms (Impact Factor: 2.77). 06/2010; 25(3):228-32. DOI: 10.1177/0748730410368016
Source: PubMed

ABSTRACT

The use of luciferase reporter genes together with luminescence detection has enabled high frequency monitoring of molecular circadian clock function in living tissues. With the help of an intensified CCD camera combined with an inverted epifluorescence microscope, the authors have established a new imaging strategy that makes use of transgenic cell type-specific expression of fluorescent proteins to identify cells of interest for subsequent circadian luminescence recording at single-cell resolution.

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Available from: Michael T. Sellix, Mar 17, 2014
    • "Measuring the circuit-wide response to light is feasible in Drosophila because the entire fly brain can be cultured [9] and approximately half the neurons in the fly circadian circuit autonomously express the blue light receptor Cryptochrome (CRY) [10, 11] , which provides the primary mechanism for light resetting the circadian clock and acute light-evoked increases in firing rate in circadian neurons [12, 13] . To address how light reorganizes the activity of the Drosophila circadian circuit mapped at single-cell resolution, we developed a culture system for Drosophila adult whole brains [9], then refined and combined high-resolution imaging of circuit-wide single oscillators [14, 15] with sophisticated mathematical analytical tools [16, 17]. For in vivo comparison, we performed anti-PER ICC using the same light/dark protocols used for whole-brain imaging. "
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