Feasibility study of incident dark-field video microscope for measuring microcirculatory variables in the mouse dorsal skinfold chamber model
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Background: Despite the importance of microcirculation in organ function, monitoring microcirculationis not a routine practice. With developments in microscopic technology, incidentdark field (IDF) microscopy (Cytocam) has allowed visualization of the microcirculation. Dorsal skinfold chamber (DSC) mouse model has been used to investigate microcirculationphysiology. By employing Cytocam-IDF imaging with DSC model to assess microcirculatoryalteration in lipopolysaccharide (LPS)-induced endotoxemia, we attempted to validate availabilityof Cytocam-IDF imaging of microcirculation. Methods: DSC was implanted in eight BALB/c mice for each group; control and sepsis. Bothgroups were given 72 hours to recover from surgery. The sepsis group had an additional 24-hour period of recovery post-LPS injection (4 mg/kg). Subsequently, a video of the microcirculationwas recorded using Cytocam. Data on microcirculatory variables were obtained. Electronmicroscopy was implemented using lanthanum fixation to detect endothelial glycocalyxdegradation. Results: The microcirculatory flow index was significantly lower (control, 2.8±0.3; sepsis,2.1±0.8; P=0.033) and heterogeneity index was considerably higher (control, 0.10±0.15;sepsis, 0.53±0.48; P=0.044) in the sepsis group than in the control group. Electron microscopyrevealed glycocalyx demolishment in the sepsis group. Conclusions: Cytocam showed reliable ability for observing changes in the microcirculationunder septic conditions in the DSC model. The convenience and good imaging quality andthe automatic analysis software available for Cytocam-IDF imaging, along with the ability toperform real-time in vivo experiments in the DSC model, are expected to be helpful in futuremicrocirculation investigations.
Abstract
Background: Despite the importance of microcirculation in organ function, monitoring microcirculationis not a routine practice. With developments in microscopic technology, incidentdark field (IDF) microscopy (Cytocam) has allowed visualization of the microcirculation. Dorsal skinfold chamber (DSC) mouse model has been used to investigate microcirculationphysiology. By employing Cytocam-IDF imaging with DSC model to assess microcirculatoryalteration in lipopolysaccharide (LPS)-induced endotoxemia, we attempted to validate availabilityof Cytocam-IDF imaging of microcirculation. Methods: DSC was implanted in eight BALB/c mice for each group; control and sepsis. Bothgroups were given 72 hours to recover from surgery. The sepsis group had an additional 24-hour period of recovery post-LPS injection (4 mg/kg). Subsequently, a video of the microcirculationwas recorded using Cytocam. Data on microcirculatory variables were obtained. Electronmicroscopy was implemented using lanthanum fixation to detect endothelial glycocalyxdegradation. Results: The microcirculatory flow index was significantly lower (control, 2.8±0.3; sepsis,2.1±0.8; P=0.033) and heterogeneity index was considerably higher (control, 0.10±0.15;sepsis, 0.53±0.48; P=0.044) in the sepsis group than in the control group. Electron microscopyrevealed glycocalyx demolishment in the sepsis group. Conclusions: Cytocam showed reliable ability for observing changes in the microcirculationunder septic conditions in the DSC model. The convenience and good imaging quality andthe automatic analysis software available for Cytocam-IDF imaging, along with the ability toperform real-time in vivo experiments in the DSC model, are expected to be helpful in futuremicrocirculation investigations.
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