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Example sentences for: rhod-
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The time to achieve a stable state (particularly with respect to effluent PO 2 concentrations) or load rhod-2, and then acquire baseline data and perform 1 intervention is 60-90 minutes, beyond which the preparation is not stable and so more than 1 intervention per mouse heart was not undertaken.
For rhod-2 the value of a is approximately 0, thus for rhod-2, F 0 was assumed to be equal to F b .
Quantification of the relative amount of rhod-2 in the heart using absorbance measurements, was done by taking the ratio of absorbance at 524 nm (rhod-2 sensitive) to 589 nm (rhod-2 insensitive) which eliminated the effect of motion as both wavelengths would be equally affected by motion, though only 524 reflected the concentration of rhod-2 [ 7 ] . These wavelengths (524, 589 nm) were chosen as these were isosbestic points not effected by changes in absorbance of myoglobin induced by oxygen desaturation [ 7 ] . In solution maximal rhod-2 absorbance is at 554 nm.
where K d is the dissociation constant for rhod-2 and calcium (determined by in-vitro calibration with rhod-2 and myoglobin by del Nido et al, ref [ 20 ] , and confirmed by in-vivo manganese quenching, ref [ 29 ] ) and is 710 nM, F t = fluorescence at time t, F max =maximal fluorescence from tetanized heart, and the fluorescence from the heart assuming rhod-2 had no calcium bound is given by F 0 = F b + a (F max - F b ), where F b is the background counts from the heart prior to dye loading, and a = rhod-2 fluorescence in the absence of calcium / rhod-2 fluorescence in the presence of saturating calcium.
Isolated perfused mouse heart measurements of intracellular calcium and energetics have been performed using other methods by other investigators [ 17 18 ] . The unique aspects of the present study are that we use the calcium sensitive fluorescent dye rhod-2 to measure intracellular calcium [ 6 7 ] , and that we have related these measurements to both myocardial oxygen consumption and developed pressure.
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