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. 2015 May 1;4:e04810. doi: 10.7554/eLife.04810

Figure 1. Defining the thermal range and quantifications.

(A and B) Progeny tests were performed on acclimated C. elegans worms from 7.5°C to 27°C and C. briggsae worms from 9°C to 30°C. Dotted line highlights 90% embryonic viability. Temperatures below 20°C exhibiting less than 90% viability are shown in cyan, temperatures above 20°C exhibiting less than 90% viability in magenta. Between panels A and B, we show the thermal range of each species. Error bars show SEM. (C–F) Stills from a time-lapse temperature-controlled DIC microscopy recording of a first-cell stage embryo at the indicated stages (G–J) Examples of feature quantification at the different cellular stages (24°C): female pronucleus speed (G), pronuclei position during centration-rotation (H), spindle pole oscillations (I), as well as areas of the AB (anterior) and P1 (posterior) daughter cells (J). See ‘Materials and methods’ for details on the quantifications. Figure 1—figure supplement 1 shows the temperature control setup. Figure 1—source data 1 lists all the quantified features and their thermal response within and beyond the thermal range.

DOI: http://dx.doi.org/10.7554/eLife.04810.003

Figure 1—source data 1. Quantified features.
List of features that were quantified and their thermal responses within and beyond the thermal range for C. elegans (N2). Within the thermal range, features were categorized as ‘temperature-dependent’ if the Pearson correlation p-value was below 0.0014 = 0.05/35 (see ‘Materials and methods’ for Bonferroni correction; ‘temperature-independent’ is shown underlined). Beyond the thermal limit, we performed an F-test to determine if the thermal response of the feature was changing compared to within the thermal range (see ‘Materials and methods’; we indicated a change in thermal response when the F-test p-value was below 0.0014, highlighted in bold). Abbreviations: PC: pseudo-cleavage, PM: pronuclear meeting, ME: mitotic entry, T: temperature, C/R: centration-rotation, MT: microtubules. The following features were also quantified but displayed no consistent thermal response both within and beyond the thermal range and hence were not included in the table: anterior-most position at the end of C/R, number of anterior and posterior oscillations, spindle position at the onset of oscillations.
DOI: http://dx.doi.org/10.7554/eLife.04810.004
elife04810s001.docx (100.9KB, docx)
DOI: 10.7554/eLife.04810.004

Figure 1.

Figure 1—figure supplement 1. Temperature-control setup.

Figure 1—figure supplement 1.

(A) The temperature within the sample was measured using a thin thermocouple of type K (see ‘Materials and methods’), connected to a temperature controller. This temperature feedback was used to control the temperature of the air blown on the sample and objective, so that sample and set temperatures always matched. (B) We verified that the temperature controller was well calibrated over the range of temperatures of interest by checking the temperature on the sample with yet another thermocouple connected to a separate external thermometer at the same time. We report the measured temperature on the sample by the temperature controller and external thermometer. The measured imprecision was of the order of 0.2°C at all temperatures.
DOI: http://dx.doi.org/10.7554/eLife.04810.005