A differential scanning calorimeter for ice nucleation distribution studies--application to bacterial nucleators.

A differential scanning calorimeter has been developed for the automatic detection and measurement of dropwise freezing within a sample of 100-200 water drops. A typical drop size of 1 microliter is employed. The sample is distributed on flat, square (4-cm) thermoelectric sensors and the temperature is scanned downward by conductive cooling to a liquid nitrogen bath. The rate of cooling, typically 1 degree C/min, is set by the choice of a heat conduction rod between the calorimeter and the liquid nitrogen bath. The voltages from the thermopiles along with a system temperature-measuring thermocouple are continuously monitored by digital voltmeters and recorded every half-second in a computer memory. A freezing event in a drop is detected by a characteristic voltage signal whose integral with time is proportional to the size of the drop and its heat of fusion. The half-life of a freezing event signal is 10 s for a 1-microliter drop. The integrated signal produced from multiple freezing events is shown to provide a direct measure of the number of drops frozen at a given temperature. A distribution curve and its smoothed derivative can be constructed directly from these measurements. The instrument, which is termed an "ice nucleometer," is illustrated in determining the ice nucleation distribution in a population of Escherichia coli harboring cloned ice nucleation genes.