Measurement of colloid osmotic pressure in submicrolitre samples

A colloid osmometer for submicrolitre samples was constructed from solid polymethylmetacrylate and acrylnitrilmethylmetacrylate blocks, exposing a 0.85 mm diameter area of a Diaflo PM-30 ultrafiltration membrane. The unknown sample, contained in a 1-microliter glass micropipette, was applied to the membrane by suction, providing minimal exposure to air. The lower limit for successful application was 0.1-0.2 microliter. The accuracy of colloid osmotic pressure (COP) measurement depends strongly on the effective compliance of the pressure transducer. We tested three different systems: (i) A Hewlett-Packard 1280 'medical' transducer gave acceptable measurements on 1-microliter samples. In smaller samples (0.1-0.5 microliter) COP was underestimated, especially at COP greater than 10 mmHg. The equilibration time was 10-30 min. (ii) As (i), but with air pressure applied to the sample by a servoregulated pump, minimizing fluid transport through the membrane. Accurate measurements on 0.2-microliter samples were obtained in the course of 2-3 min, but the system required special instrumentation and some operating experience. (iii) An 'industrial' transducer, SensoNor AE-88o, with very low compliance, gave accurate measurements in the course of 1-3 min on samples as small as 0.1-0.2 microliter and COP up to 37 mmHg. We recommend system (iii) for samples smaller than 1 microliter.