Figures & data
Figure 1 Water replacement hypothesis.
Notes: The diagram shows how trehalose is thought to stabilize lipid bi-layers during dehydration. Adapted from Arch Biochem Biophys, 242(1). Crowe LM, Crowe JH, Rudolph A, Womersley C, Appel L. Preservation of freeze-dried liposomes by trehalose. 240–247. Copyright 1985, with permission from Elsevier.Citation44
Figure 2 A possible method to test tolerance to inoculative freezing in insects.
Notes: The freeze-tolerance assay is conducted in a programmable thermostat with adjustable cooling rate. (A) The temperature program: 1) hold for 10 minutes at 0°C (manipulation time; tubes with test subjects and ice crystals are inserted into the thermostat), 2) cool to T at a rate of 0.1°C/min or slower/faster, 3) hold X min at T, 4) heat to +5°C at a rate of 0.1°C/min or slower/faster. Arrow marks the onset of freezing of the water contained in moistened cellulose (see B for details). Double arrow indicates freezing exotherm – a confirmation that the test subject was successfully inoculated. (B) Test tube. The test subject is put on a piece of moistened cellulose (gray rectangle) and covered with another piece of moistened cellulose of the same size and water content. The subject is immobilized by being gently pressed between the two pieces of cellulose as well as by low temperature in the thermostat (pre-cooled to 0°C). Asterisk represents the ice crystal, put inside the tube in order to initiate freezing. Vertical line with circle at lower end represents a thermocouple, mounted between the two pieces of cellulose in order to record if inoculation occurred. Based on data from Koštál et alCitation101 and Rozsypal et al.Citation97
