Supplementary MaterialsFigure S1: Using the SMR to measure the buoyant mass of a cell in H2O and D2O. angle can be shown to map directly to density. c) Contour map showing cell mass BIIB021 inhibition as a function of two buoyant masses. This function is usually linear, with a gradient oriented to the lower right (higher BIIB021 inhibition buoyant mass in H2O, lower buoyant mass in D2O).(TIF) pone.0067590.s003.tif (861K) GUID:?F84FA47E-7288-4AB2-BDF3-C7C4DB8F10D3 Figure S4: Comparison of measured BIIB021 inhibition data (solid lines) to simulations of buoyant mass measurement errors propagating through the density calculation for samples. Dashed lines show expected dry density distributions assuming all cells have the same density and that density is the median observed dry density (vertical line).(TIF) pone.0067590.s004.tif (652K) GUID:?13B13D23-582B-41C2-AE86-BC953C4F8785 Figure S5: Dry density distributions for budded and unbudded yeast cells, by timepoint. P-values are for two-sided Mann-Whitney U assessments.(TIF) pone.0067590.s005.tif (338K) GUID:?0D6294CC-C750-4A54-A13B-A313341B0A45 Physique S6: Contour plots of dry density estimates when the buoyant mass measurements arent made in real H2O or real D2O. Intracellular water fractions are in fraction of total volume. Dashed line shows equal departure (in density) from real fluids. Pure H2O and 91 (v/v) D2O:H2O densities are the red dot in the lower left corner of each Rabbit polyclonal to ADAMTSL3 figure, of which stage the dry density correctly is calculated. As salts (or various other impermeable elements) are put into the fluid, it becomes more dense as well as the intracellular drinking water is zero neutrally buoyant longer. This introduces organized error in to the dried out thickness dimension, which depends on how much of the cell is usually water. The measurements weve made using 1 PBS in both fluids are shown as black dots.(TIF) pone.0067590.s006.tif (781K) GUID:?5FB26577-13AD-4D96-AA13-A6E9E911E748 Figure S7: Time between measurements (exposure time) calculated dry density for single cells in each of nine analyses of samples (2C3 technical replicates for each of 4 samples). Assuming the cell was nearly immediately immersed in D2O after the first measurement, this should be a good approximation of time spent in D2O. Collection shows regular least squares fits, which agreed well with strong fits (Huber weights). Correlations are all statistically insignificant at ?=?0.05 (?=?0.006 for each test, using Bonferroni correction). P-values are given for slope being non-zero using one-sided t-test.(TIF) pone.0067590.s007.tif (668K) GUID:?E0B7BE30-4F95-4192-A915-D64B8F4E29E6 Physique S8: Time between measurements (exposure time) calculated dry density for single cells in four experiments. Collection shows regular least squares fits, which never account for more than 5% of the total variance. Because these experiments were carried out three-channel devices, much more precise control over exposure time could be achieved, and this parameter was deliberately varied, yielding the discrete occasions seen above. Only 1 experiment showed a substantial correlation ( statistically?=?0.05/4?=?0.0125 using Bonferroni correction). P-values receive for slope getting nonzero using one-sided t-test.(TIF) pone.0067590.s008.tif (455K) GUID:?251EB387-EE1D-4890-9B04-9903BCA501D5 Document S1: Supplemental discussion: error sources, evidence for complete fluid exchange, explanation of water-content dimension remarks and technique on the need of BIIB021 inhibition single-cell measurements.(PDF) pone.0067590.s009.pdf (136K) GUID:?BD78E225-C477-4224-A096-301C9622061C Abstract a way is normally presented by all of us for immediate non-optical quantification of dried out mass, dried out water and density mass of one living cells in suspension. Dry out mass and dried out thickness are obtained concurrently by calculating a cells buoyant mass sequentially within an H2O-based liquid and a D2O-based liquid. Fast exchange of intracellular H2O for D2O.