The present study investigated the relationship between blood and subcutaneous interstitial fluid (ISF) glucose by employing an amperometric glucose sensor specifically developed for 3-day continuous glucose monitoring. The apparent sensor sensitivity and ISF glucose equilibration delay were estimated on separate days during hyperglycemic clamps in four dogs in which insulin was either suppressed with somatostatin, allowed to change, or increased with an exogenous infusion. A 2-h sensor "settling-in" period was allowed before the clamps. During insulin deficiency, the sensor sensitivity and ISF glucose delay were 0.23 +/- 0.03 nA per mg/dl and 4.4 +/- 0. 8 min. Sensitivity was not affected by increases in endogenous (0.30 +/- 0.04 vs. 0.28 +/- 0.04 nA per mg/dl) or exogenous insulin (0.18 +/- 0.01 vs. 0.16 +/- 0.01 nA per mg/dl) nor was the delay (3.3 +/- 1.2 vs. 5.7 +/- 1.1 and 9.2 +/- 2.6 vs. 12.3 +/- 1.7 min; P > 0.05 for all). Sensor glucose accurately predicted plasma glucose without correcting for delays <10 min (r > 0.9 for all), whereas for longer delays a digital corrective filter was used (r = 0.91 with filter). We conclude that the relationship between blood and ISF glucose is not affected by insulin and that delays in ISF glucose equilibration can be corrected with digital filters.