4.7 Article

Accurate and online quantification of viable Rhodobacter sphaeroides cells using a flow cytometry-dielectric spectroscopy (FCM-DS) method

期刊

TALANTA
卷 245, 期 -, 页码 -

出版社

ELSEVIER
DOI: 10.1016/j.talanta.2022.123448

关键词

Rhodobacter sphaeroides; Flow cytometry; Dielectric spectroscopy; Viable cell count; Coenzyme Q10; FCM-DS method

资金

  1. National Key Research and Development Program of China [2017YFF0204602, 2018ZX10734404]
  2. Fundamental Research Funds for Central Public Welfare Scientific Research Institutes
  3. National Institute of Metrology, P.R. China [AKYZD2110, AKY1818, AKY1958]
  4. Science and Technology Program of State Administration for Market Regulation [2019MK114]

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The study developed an integrated method using flow cytometry and dielectric spectroscopy to accurately quantify viable cells and their capacitance value online. The results suggest that this method can help devise precise fermentation control strategies.
Accurate and online quantification of viable cells is one of the necessary requirements during the microbial fermentation process for high productivity. The flow cytometry (FCM)-based method accurately quantifies viable cells, but this offline method cannot reflect the counts constantly. The dielectric spectroscopy (DS) sensor is widely utilized to monitor viable cells online; however, accurately converting the capacitance value of the DS sensor to the viable bacterial cell counts has barely been tried. We have developed a method by coupling the principles and techniques of FCM and the DS sensor to quantify viable Rhodobacter sphaeroides cells. Using specific fluorescent antibodies and propidium iodide (PI), viable R. sphaeroides cells were accurately quantified within 30 min by FCM. The DS sensor was combined with the FCM to create a direct capacitance-viable cell count quantification system. The LOD (limit of detection) of the FCM-DS method was 8 x 10(8) CFU/mL, RSD (relative standard deviation) < 5%, along with good reproducibility of the results. Finally, the viable cell count, obtained from the FCM-DS method, was applied to regulate the specific oxygen uptake rate (Q(O2)) that increased the production of coenzyme Q10 by 8.1%. Together, our results strongly suggest that viable cells can be accurately quantified online by the integrated FCM-DS method, which would help to devise precise fermentation control strategies.

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