Journal: Frontiers in Microbiology

Article Title: Single-Cell Growth Rates in Photoautotrophic Populations Measured by Stable Isotope Probing and Resonance Raman Microspectrometry

doi: 10.3389/fmicb.2017.01449

Figure Lengend Snippet: Examples of resonance Raman spectra obtained from EUBMIX-hybridized (Cy3) Synechococcus sp. (RS9916) cells (0.5–1.8 μm in diameter) pictured in fluorescence (Y3 ET, k—545 excitation/610 emission) (A) and bright-field (B) images. Cells manually targeted in the microscope field by the computer mouse are numbered, then automatically revisited under laser illumination for Raman interrogation. (C) Single-cell spectrum i was obtained from a culture grown on natural 13 C abundances ( f media = 0.011 DI 13 C). Single-cell spectra ii–v were obtained from a culture in f/2 media augmented with 2 mM bicarbonate yielding a final fractional 13 C-content of 54% (0.54 f media ) after 3 ( ii ), 6 ( iii ), 12 ( iv ), and 24 ( v ) days of incubation. Vertical lines denote major peak positions in the control culture. Spectra were baseline-corrected, intensities normalized from 0 to 1, and smoothed using standard Renishaw™ Wire 4.1® routines.

Article Snippet: Graphics were produced by either Renishaw® Wire 4.1™ or SigmaPlot™ 13.0 software.

Techniques: Fluorescence, Microscopy, Incubation

Journal: Frontiers in Microbiology

Article Title: Single-Cell Growth Rates in Photoautotrophic Populations Measured by Stable Isotope Probing and Resonance Raman Microspectrometry

doi: 10.3389/fmicb.2017.01449

Figure Lengend Snippet: Examples of the varying contributions of 12 C= 12 C, 12 C= 13 C, and 13 C= 13 C isotopologues to the shape, position, and areas of the ν (C = C) Raman spectral peak for carotenoids of cells assimilating varying amounts of DI 13 C. Each Raman spectrum was obtained from individual cells grown in either 0.011 f media (A) or 0.54 f media for 3 (B) , 6.2 (C) , or 9.2 (D) days. Spectra were subjected to local baseline subtraction (1,360–1,660 cm −1 ), intensity normalization (0–1), and a full Voigt curve-fitting routine (convolution of Lorentzian and Gaussian profiles) with 5,000 iterations or a 0.00001 tolerance using Renishaw™ Wire 4.1® software. Center positions for each of the three isotopologues were constrained within narrow consensus ranges (± 1 cm −1 ) determined from regression coefficients presented in Figure . Isotopologue peak widths and symmetries were allowed to float to optimize curve fits.

Article Snippet: Graphics were produced by either Renishaw® Wire 4.1™ or SigmaPlot™ 13.0 software.

Techniques: Software