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. Author manuscript; available in PMC: 2012 Jul 10.
Published in final edited form as: Nat Methods. 2012 Jan 22;9(3):273–276. doi: 10.1038/nmeth.1857

Figure 1.

Figure 1

Selective labeling of artery walls by Alexa Fluor 633. (a,b) Two-photon microscopy image of the mouse visual cortex after a 4 pounds per square inch (p.s.i.) 1-s puff of Alexa Fluor 633 from a micropipette (a; + indicates tip position) and an autofluorescence image (b) obtained simultaneously in vivo using 2 mW laser power. (c,d) Image from the rat visual cortex after a 7 p.s.i. 1-s puff of Alexa Fluor 633 in vivo (c) and fluorescence image of the same cortical site taken after intravenous injection of fluorescein dextran (d). (e,f) Images of arteriole walls in mouse visual cortex after intravenous injection of Alexa Fluor 633 (e) and of the same site after injection of fluorescein dextran (f). (g) Average intensity 66 μm z-dimension projection image from mouse visual cortex after an intravenous injection of fluorescein dextran and local pipette injection of Alexa Fluor 633. (h) Dilation of an Alexa Fluor 633–labeled arteriole from the rat visual cortex in response to drifting grating visual stimuli (gray bar). The intersection between the regression line (dotted green) and zero level (dashed black) represents the latency of dilation (0.8 s). Purple shading indicates s.e.m. (n = 16 trials). (i) Molecular structures of Alexa Fluors 633 and 594. (j) Relative brightness of arteriole walls labeled with Alexa Fluors 594, 633 and 647. Data were pooled from five experiments in the rat cortex (error bars, s.e.m.). (k) Image of an immunostained section from the macaque monkey neocortex using an antibody to α-SMA and showing Alexa Fluor 633 labeling. (l) Image of an immunostained section of mouse neocortex with an antibody to GLUT1 and showing Alexa Fluor 633 labeling. (m) Image of a mouse aorta labeled with Alexa Fluor 633 and collected by using 2 mW two-photon laser power. (n) Image of a mouse femoral artery stained with an antibody to laminin and showing Alexa Fluor 633 labeling. In vivo images (a–f) were not background-subtracted or movement-corrected. Images shown in k, l and n were collected with sequential scanning confocal microscopy. Scale bars, 100 μm (a–g), 10 μm (k,l,n) and 50 μm (m).

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