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Figure 6

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Techniques for automated and multiscale microscopy. (a) Automated microscopy. Neurons were cotransfected with a morphological marker, mRFP (red), and a mutant huntingtin protein fused to GFP, Httex1-Q47-GFP (green). The expression levels of mutant huntingtin and cell fates were tracked automatically in thousands of neurons over multiple days (Arrasate et al. 2004). (b) Multiscale microscopy as illustrated in a series of increasingly magnified images of a medium spiny neuron from mouse nucleus accumbens. (Left) Mosaic of low-magnification, transmitted light images of mouse striatum. (Left inset) Contrast-enhanced, close-up of an individual neuron. (Middle) Two-photon fluorescence image of the same neuron labeled with Lucifer Yellow and Alexa Fluor 568. (Right) Tomographic reconstruction of a dendrite by electron microscopy and the corresponding segmentation of dendritic spines from the same neuron (images courtesy of E. Bushong, M. Martone, and M. Ellisman, available online at the Cell Centered Database, http://ccdb.ucsd.edu). (c) Array tomography. Volumetric image of mouse vibrissa somato-sensory cortex. The image reveals synapses labeled by rhodamine-conjugated antisynapsin I (small red objects), cellular nuclei (DAPI-DNA, blue), autofluorescence from blood cells (large red objects), and a subset of pyramidal neurons labeled by FITC-conjugated anti-GFP (green) and was rendered from 134 sections, each 200-nm thick (image courtesy of K. Micheva and S. Smith; Micheva & Smith 2007). (d) All-optical histology. Maximum intensity projections of fluorescently labeled vasculature in the mouse vibrissa somato-sensory cortex. A penetrating arteriole is highlighted in yellow (P.S. Tsai, P. Blinder, J.A. Kaufhold, B. Friedman, and D. Kleinfeld, unpublished data).

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