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Review
. 2004 Oct 20;24(42):9223-7.
doi: 10.1523/JNEUROSCI.3344-04.2004.

Novel approaches to monitor and manipulate single neurons in vivo

Michael Brecht  1 , Michale S Fee, Olga Garaschuk, Fritjof Helmchen, Troy W Margrie, Karel Svoboda, Pavel Osten
Affiliations
Review

Novel approaches to monitor and manipulate single neurons in vivo

Michael Brecht et al. J Neurosci. .
No abstract available

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Figures

Figure 1.
Figure 1.
Monitoring neurons with improved cellular resolution. A, Long-term in vivo imaging of GFP-expressing axons in the adult mouse barrel cortex. Note gained (e.g., green arrowhead), lost (blue arrowhead), and stable (yellow arrowheads) putative synaptic terminals (A. Holtmaat, personal communication). B, Ca2+ transients evoked by whisker deflection. Left, A high-magnification image of layer 2/3 neurons in vivo (depth, 130 μm) in the barrel cortex of a 13-d-old mouse. Right, Line-scan recordings of Ca2+ transients evoked in two neurons by a deflection of the majority of whiskers on the contralateral side of the mouse's snout. The position of the scanned line and the cells analyzed are indicated (left) (modified from Stosiek et al., 2003).
Figure 2.
Figure 2.
Monitoring neural activity under natural conditions. A, Top, Overview of motorized microdrive. Electrodes are driven independently by small brushless DC motors. Bottom, Sample recording from a single neuron in the nucleus robustus archistriatalis of a zebra finch during singing (adapted from Fee and Leonardo, 2001). B, Top, Principal design of the miniaturized two-photon microscope. Excitation light and fluorescence emission light are guided through optical fibers. The two-photon fiberscope is mounted above a small cranial window and can be carried by freely moving rats. Bottom, Two-photon fiberscope-acquired image of a layer 2 neuron filled with Calcium Green-1 (left) and small blood vessels labeled via tail-vein injection of FITC-dextran (right) (adapted from Helmchen et al., 2001).
Figure 3.
Figure 3.
Targeting and manipulating single neurons. A, Top, Targeted whole-cell recording from a calretinin-positive interneuron in the glomerulus layer of the mouse olfactory bulb (postnatal day 21). Image is an overlay of two channels (red and green) showing the illuminated patch pipette filled with Alexa 594 attached to the GFP-labeled interneuron. Bottom, Trace shows membrane potential in response to odor presentation (2 sec). B, Enhanced GFP-expressing lentivirus was injected into the parenchyma of rat layer 2/3 somatosensory cortex (postnatal day 28). After a 2 week expression period, the rat was killed, and enhanced GFP fluorescence was visualized from 100 μm paraformaldehyde-fixed brain sections. The image is a maximal projection of 40 confocal sections, separated by 2 nm. C, Intracellular stimulation of the rat primary whisker motor cortex. Top trace, Position of whisker E1 during an intracellular stimulation trial. Bottom trace, Membrane potential recordings during current injection (10 action potentials at 50 Hz); the stimulation current is not shown.
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