Microvascular geometry and differential permeability in the eye during inflammation revealed with dual channel multiphoton microscopy

Bateman, Ryon M.; Hodgson, Kevin; Breemen, C. van; Walley, Keith R.

doi:10.1117/12.647309

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“…This contrasts with the sensitivity of the mouse eye, where we observed that powers above a few milliwatts induced hyperemia. 18 While we confirmed that QDs are taken up by the liver and spleen, 15,19 we also observed that mPEGylated 5000-MW QDs aggregated at microvascular bifurcations, suggesting that hemodynamic forces play a role in the biodistribution of QDs, presumably by increasing the contact time between nanoparticles and rolling or adhered leukocytes. This microvascular localization of QDs likely explains why the circulating halflife of QDs assessed by whole body noninvasive imaging was found to be twice that determined by venipuncture.…”

Section: Discussionsupporting

confidence: 72%

Endotoxemia increases the clearance of mPEGylated 5000-MW quantum dots as revealed by multiphoton microvascular imaging

et al. 2007

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Imaging the microcirculation is becoming increasingly important in assessing life-threatening disease states. To address this issue in a highly light absorbing and light scattering tissue, we use laser scanning multiphoton microscopy and fluorescent 655-nm 5000-MW methoxy-PEGylated quantum dots to image the functional microcirculation deep in mouse hind limb skeletal muscle. Using this approach, we are able to minimize in vivo background tissue autofluorescence and visualize complete 3-D microvascular units, including feeding arterioles, capillary networks, and collecting venules to depths of 150 to 200 microm. In CD1 mice treated with lipopolysaccharide to model an endotoxemic response to bacterial infection, we find that these quantum dots accumulate at microvascular bifurcations and extravasate from the microcirculation in addition to accumulating in organs (liver, spleen, lung, and kidney). The quantum dots are cleared from the circulation with a first-order elimination rate constant seven times greater than under normal conditions, 1.6+/-0.06 compared to 0.23+/-0.05 h(-1), P<0.05, thereby reducing the imaging time window. In vitro experiments using TNFalpha treated isolated leukocytes suggest that circulating monocytes (phagocytes) increased their nonspecific uptake of quantum dots when activated. In combination with multiphoton microscopy, quantum dots provide excellent in vivo imaging contrast of deep microvascular structures.

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Section: Discussionsupporting

confidence: 72%