Among the new molecular tools available to scientists and engineers, some of the most useful incl... more Among the new molecular tools available to scientists and engineers, some of the most useful include fluorescently tagged biomolecules. Tools, such as green fluorescence protein (GFP), have been applied to perform semi-quantitative studies on biological signal transduction and cellular structural dynamics involved in the physiology of healthy and disease states. Such studies focus on drug pharmacokinetics, receptor-mediated endocytosis, nuclear mechanobiology, viral infections, and cancer metastasis. In 1976, fluorescence recovery after photobleaching (FRAP), which involves the monitoring of fluorescence emission recovery within a photobleached spot, was developed. FRAP allowed investigators to probe two-dimensional (2D) diffusion of fluorescently-labelled biomolecules. Since then, FRAP has been refined through the advancements of optics, charged-coupled-device (CCD) cameras, confocal microscopes, and molecular probes. FRAP is now a highly quantitative tool used for transport and ki...
Purpose-Laser Doppler velocimetry provides a flow to the skin. The purpose of our studies is to m... more Purpose-Laser Doppler velocimetry provides a flow to the skin. The purpose of our studies is to measure of relative skin perfusion only, and cannot compare LDV measurements of skin blood flow as yet be related to any absolute measure of blood with quantitative perfusion fluorometry and our
We have constructed a confocal scanner suitable for routine microarray analysis from commercially... more We have constructed a confocal scanner suitable for routine microarray analysis from commercially available parts. We have outlined the details that should be considered when designing such an instrument and listed some of the specific components comprising the system [the full list of system components is available on CD from the corresponding author (D.J.G.) at no charge]. Here, we describe the methods used to test the linearity and sensitivity of the instrument. Performance was evaluated with two commonly used dyes, fluorescein and Cy5. While the instrument had a linear correlation between the dye concentration and fluorescence intensity, the observed deviation from a slope of 1.0 underscores the importance of running multipoint calibration experiments to obtain accurate dye quantitation over the full dynamic range of the scanner. This method has utility in testing commercial instruments in addition to the scanner described here. An array with over 300 spots dyed with Cy3 was sca...
Journal of applied physiology (Bethesda, Md. : 1985), 1985
The flux of He and O2 through intact adult human skin was measured at various inspired concentrat... more The flux of He and O2 through intact adult human skin was measured at various inspired concentrations and skin temperatures. The skin surface was then stripped with cellophane tape to alter the diffusional conductance of the stratum corneum. He flux for stripped skin was used to estimate skin perfusion as a function of local temperature, and diffusional conductance for O2 was estimated from O2 flux and perfusion. The flux of He or O2 at constant skin temperature can be related to inspired concentration by a simple linear model. Increasing surface temperature in the range 33-43 degrees C produced a much larger increase in O2 flux than in He flux for intact skin. Skin stripping greatly increased skin O2 flux. Estimated skin conductance for O2 showed a more marked temperature dependence than estimated skin perfusion. The results suggest that raising skin temperature in the range 38-43 degrees C has only a modest effect on skin perfusion and that stratum corneum conductance may have a m...
Journal of applied physiology (Bethesda, Md. : 1985), 1989
We present an analytic method for determining the effects of skin perfusion--vasculature and flow... more We present an analytic method for determining the effects of skin perfusion--vasculature and flow rates--on the flux of inert gases through human skin. We systematically specify the underlying blood flow and examine the resulting fluxes of several gases, allowing for the appropriate tissue resistances. For physiological flows, the stratum corneum has an effect equivalent to a series resistance. Helium flux at low total flow depends primarily on subdermal perfusion, but at higher flow, middermal and subpapillary effects become important. The fluxes of less permeable gases, such as argon and xenon, depend on middermal and subpapillary flow at lower total flows. From any single measurement of gas flux, it is difficult to establish an unambiguous value for the underlying blood flow, but the simultaneous measurement of different gases narrows the range of plausible conditions.
Among the new molecular tools available to scientists and engineers, some of the most useful incl... more Among the new molecular tools available to scientists and engineers, some of the most useful include fluorescently tagged biomolecules. Tools, such as green fluorescence protein (GFP), have been applied to perform semi-quantitative studies on biological signal transduction and cellular structural dynamics involved in the physiology of healthy and disease states. Such studies focus on drug pharmacokinetics, receptor-mediated endocytosis, nuclear mechanobiology, viral infections, and cancer metastasis. In 1976, fluorescence recovery after photobleaching (FRAP), which involves the monitoring of fluorescence emission recovery within a photobleached spot, was developed. FRAP allowed investigators to probe two-dimensional (2D) diffusion of fluorescently-labelled biomolecules. Since then, FRAP has been refined through the advancements of optics, charged-coupled-device (CCD) cameras, confocal microscopes, and molecular probes. FRAP is now a highly quantitative tool used for transport and ki...
Purpose-Laser Doppler velocimetry provides a flow to the skin. The purpose of our studies is to m... more Purpose-Laser Doppler velocimetry provides a flow to the skin. The purpose of our studies is to measure of relative skin perfusion only, and cannot compare LDV measurements of skin blood flow as yet be related to any absolute measure of blood with quantitative perfusion fluorometry and our
We have constructed a confocal scanner suitable for routine microarray analysis from commercially... more We have constructed a confocal scanner suitable for routine microarray analysis from commercially available parts. We have outlined the details that should be considered when designing such an instrument and listed some of the specific components comprising the system [the full list of system components is available on CD from the corresponding author (D.J.G.) at no charge]. Here, we describe the methods used to test the linearity and sensitivity of the instrument. Performance was evaluated with two commonly used dyes, fluorescein and Cy5. While the instrument had a linear correlation between the dye concentration and fluorescence intensity, the observed deviation from a slope of 1.0 underscores the importance of running multipoint calibration experiments to obtain accurate dye quantitation over the full dynamic range of the scanner. This method has utility in testing commercial instruments in addition to the scanner described here. An array with over 300 spots dyed with Cy3 was sca...
Journal of applied physiology (Bethesda, Md. : 1985), 1985
The flux of He and O2 through intact adult human skin was measured at various inspired concentrat... more The flux of He and O2 through intact adult human skin was measured at various inspired concentrations and skin temperatures. The skin surface was then stripped with cellophane tape to alter the diffusional conductance of the stratum corneum. He flux for stripped skin was used to estimate skin perfusion as a function of local temperature, and diffusional conductance for O2 was estimated from O2 flux and perfusion. The flux of He or O2 at constant skin temperature can be related to inspired concentration by a simple linear model. Increasing surface temperature in the range 33-43 degrees C produced a much larger increase in O2 flux than in He flux for intact skin. Skin stripping greatly increased skin O2 flux. Estimated skin conductance for O2 showed a more marked temperature dependence than estimated skin perfusion. The results suggest that raising skin temperature in the range 38-43 degrees C has only a modest effect on skin perfusion and that stratum corneum conductance may have a m...
Journal of applied physiology (Bethesda, Md. : 1985), 1989
We present an analytic method for determining the effects of skin perfusion--vasculature and flow... more We present an analytic method for determining the effects of skin perfusion--vasculature and flow rates--on the flux of inert gases through human skin. We systematically specify the underlying blood flow and examine the resulting fluxes of several gases, allowing for the appropriate tissue resistances. For physiological flows, the stratum corneum has an effect equivalent to a series resistance. Helium flux at low total flow depends primarily on subdermal perfusion, but at higher flow, middermal and subpapillary effects become important. The fluxes of less permeable gases, such as argon and xenon, depend on middermal and subpapillary flow at lower total flows. From any single measurement of gas flux, it is difficult to establish an unambiguous value for the underlying blood flow, but the simultaneous measurement of different gases narrows the range of plausible conditions.
Uploads
Papers by David Graves