This description of time-dependent changes in carotenoid content inside cells of R. glutinis when submerged in culture is in agreement with that measured by Bhosale & Gadre (2001 b) using HPLC. However, carotenoid quantification based on LTRS has the following advantages over traditional methods. First, it is less time consuming. The classic extraction procedure using HPLC requires over 5 h. In contrast, it only takes about 40 min to acquire Raman

spectra from 100 cells. Second, LTRS cannot cause degradation HDAC inhibitor review or isomerization of carotenoids when using a low-power laser. Third, only a small amount of sample, for example, not more than 200 μL culture, is required for carotenoid measurement. Finally, because no organic solvent is used for LTRS, environmental pollution and health hazards can be avoided. Most of our knowledge on the microbial fermentation process has been obtained by inference from cell-population www.selleckchem.com/products/AZD6244.html level data, including information on substrate concentration, product concentration, and fermentation broth pH. However, in many cases, a population of cells has a different response to the environment due to heterogeneity within the population. The increasing need to understand individual cell behavior drives the development of single-cell analytical techniques. Of particular

importance are techniques, like the one presented in this paper, which will enable us to probe the dynamic changes within an individual cell and the intercellular variability that reveals the underlying mechanisms behind the coordination of multicellular behavior. In this work, we assessed the variation in carotenoid levels per cell over 100 single cells of R. glutinis at different time points (8, 16, 32, 48, and 64 h). Figure 4 shows 10 randomly selected Raman spectra from the 100 spectral data of R. glutinis cells at each time point and Table 2 illustrates the mean value and coefficient

of variation (CV; SD/mean) for carotenoid content inside the cells at these time points. In the lag (8 h) and early exponential phases (16 h), most cells were in rapid proliferation and had a low intracellular carotenoid content. The variation in Methocarbamol carotenoid levels of cells was significant, giving a CV value of 144% and 241%, respectively. At 32 h, most cells entered the carotenogenesis phase and the heterogeneity in carotenoid levels began to diminish, with a CV value of 63%. A further decrease of variation in the carotenoid levels of cells could be seen with the increase of the carotenoid content during the late exponential and stationary phases; the CVs were 33% and 32%, respectively. The results indicate that the carotenoid levels in individual cells in a population vary significantly, especially for the population of cells in the lag and early exponential phases. In order to estimate the carotenoid level measurement errors, we made 100 measurements on a single cell randomly selected from the sample at 64 h.