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Measuring the fermentation broth of NADH photoelectric sensor

Measuring the fermentation broth of NADH photoelectric sensor. 量測發酵液之 NADH 光電感測器. Presenter: Shih- Chieh Chao Adviser: Dr. Hung-Chi Yang Chairman: Dr. Hung-Chi Yang Date:05.08.2013. Outline. Background Introduction Purpose Methods & Materials Experiment Future works Reference.

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Measuring the fermentation broth of NADH photoelectric sensor

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  1. Measuring the fermentation broth of NADH photoelectric sensor 量測發酵液之NADH光電感測器 • Presenter: Shih-Chieh Chao • Adviser: Dr. Hung-Chi Yang • Chairman: Dr. Hung-Chi YangDate:05.08.2013

  2. Outline • Background • Introduction • Purpose • Methods & Materials • Experiment • Future works • Reference

  3. Background • Fluorescent luminous phenomenon is a light cooling. When some kind of substance at room temperature by a certain wavelength of the incident light (usually ultraviolet or X-ray) radiation, absorb light energy into excited states and decay and issued by the outgoing light. • Human tissue containing many fluorescent substances, including collagen, NADH, FAD, rhodopsin, will be excited by UV fluorescence.

  4. Background • Human tissue in the cancer process, will produce a change in the structure, such as: the cells become less dense connective tissue collagen tissue to reduce; metabolism of cancer cells than normal cells, so that the increased NADH; caused by increased blood flow absorbed rhodopsin rise. Therefore, the fluorescence spectrum of the cancer tissue and normal tissue will be different, and performance in the fluorescence spectral peak position and strength of the ebb and flow.

  5. Introduction • NADH(煙醯胺腺嘌呤二核苷酸)NADH is a widespread variety of animals, plants and humans in living cells, natural substances , and is a very important compound, was not discovered until 1905, usually with many enzymes in the body co-participate in the reaction, usually called "coenzyme“.

  6. Introduction • Cell growth and energy metabolism in the cells are required to NADH, food into energy process requires NADH participate, and in the cell to produce energy in the process, NADH plays the important role of the portable electronic, in addition to NADH is also a very important the antioxidants can protect cells from damage of the harmful substances.

  7. Introduction • NADH Excitation light of 340 ± 20nm.

  8. Introduction

  9. Introduction • The most important function of the Fermenter, is to make the microorganisms in the growth conditions it needs to reach the desired proliferation. • Therefore the design and production of the fermentation tank you need to have a lot of considerations.

  10. Introduction • Need to maintain long-aseptic operation. • Microbial metabolism of aeration and agitation, but can not give microbial mechanical shock stirring intensity. • Need to configure the device temperature system Imperial. • Need to be able to control the pH. • Need to be able to sample. • Must be appropriate size. • Internally to maintain a smooth inner surface, not have welded junction. • By fermentation of a variety of different, you need to install the necessary equipment incidental.

  11. Introduction

  12. Purpose • This study focused on : • The sensor can be fully inserted fermenter • Continuous and real-time performance measurement

  13. Purpose PC Detector MCU Control Broth + fungus 450 nm LED 340 nm

  14. Purpose LED*4 PWM 30F4011 DET A/D PC TIMER UART RS232

  15. Methods & Materials • Solidwork

  16. Methods & Materials • Type 304—the most common grade; the classic 18/8 stainless steel. Outside of the US it is commonly known as "A2 stainless steel”. • Type 316—the second most common grade (after 304); for food and surgical stainless steel uses; alloy addition of molybdenum prevents specific forms of corrosion. It is also known as marine grade stainless steel due to its increased resistance to chloride corrosion compared to type 304. 316 is often used for building nuclear reprocessing plants.

  17. Methods & Materials

  18. Methods & Materials LED Detector

  19. Methods & Materials

  20. Methods & Materials

  21. Methods & Materials 20mm 44mm 12mm 10mm 8mm 10mm 20mm 40mm 44mm 40mm

  22. Methods & Materials

  23. Methods & Materials

  24. Methods & Materials 26mm 30mm 18mm 14mm 14mm 26mm 30mm 26mm

  25. Methods & Materials • Excitation light ： NSHU551B

  26. Methods & Materials • Kupo：UV Transmission

  27. Methods & Materials • NADH emit light of450±20nm， so the selection of the received light 320-730nm. • Hamamatsu：S1133

  28. Methods & Materials • Kupo：C11715

  29. Methods & Materials • Kupo：UV Blocking

  30. Methods & Materials • UV Blocking+C11715

  31. Experiment

  32. Experiment

  33. Experiment

  34. Experiment

  35. Experiment • Fluorescein, 90+% L13251 • Formula: ：C20H12O5 • Molecular weight :332.32 • Orange powder

  36. Experiment • Deployment method, beginning first with a 1-to-1 water diluted to 47.5% alcohol with 95% alcohol(AOH)

  37. Experiment

  38. Experiment

  39. Experiment • Sigma N8129 ： • Reduced coenzyme • Yellowish white powder • Stored at -20 ℃ • Excitation light = 340 nm。 • Emit light = 460 nm。

  40. Experiment • Fluorescence Spectrometer F-7000

  41. Future works • Circuit design • 30F4011 • Labview

  42. Reference [1] Y-C Liu, F-S Wang, W-C Lee, “ On-line monitoring and controlling system for fermentation processes. “ Biochemical Engineering Journal 7 (2001) 17–25. [2] Monici M. (2005). "Cell and tissue autofluorescence research and diagnostic applications".BiotechnolAnnu.Rev.11:227–56.doi:10.1016/S1387-2656(05)11007-2. PMID 16216779. [3] Georgakoudi I, Jacobson BC, Müller MG, Sheets EE, Badizadegan K, Carr-Locke DL, Crum CP, Boone CW, Dasari RR, Van Dam J, Feld MS (2002-02-01). "NAD(P)H and collagen as in vivo quantitative fluorescent biomarkers of epithelial precancerous changes". Cancer Res. 62 (3): 682–687. PMID 11830520 [4] Windholz, Martha (1983). The Merck Index: an encyclopedia of chemicals, drugs, and biologicals (10th ed.). Rahway NJ, US: Merck. p. 909. ISBN 911910271 Check |isbn= value (help). [5] Bakker BM, Overkamp KM, van Maris AJ, et al. (2001). "Stoichiometry and compartmentation of NADH metabolism in Saccharomycescerevisiae". FEMS Microbiol. Rev. 25 (1): 15–37. [6] Nicholls DG; Ferguson SJ (2002). Bioenergetics 3 (1st ed.). Academic Press. ISBN 0-12-518121-3. [7] Sistare FD, Haynes RC (15 October 1985). "The interaction between the cytosolic pyridine nucleotide redox potential and gluconeogenesis from lactate/pyruvate in isolated rat hepatocytes. Implications for investigations of hormone action". J. Biol. Chem. 260 (23): 12748–53. PMID 4044607.  [8] Freitag A, Bock E (1990). "Energy conservation in Nitrobacter". FEMS MicrobiologyLetters 66 (1–3): 157–62. doi:10.1111/j.1574-6968.1990.tb03989.x. [9] Starkenburg SR, Chain PS, Sayavedra-Soto LA, et al. (2006). "Genome Sequence of the Chemolithoautotrophic Nitrite-Oxidizing Bacterium Nitrobacterwinogradskyi Nb-255". Appl. Environ. Microbiol. 72 (3): 2050–63. doi:10.1128/AEM.72.3.2050-2063.2006. PMC 1393235. PMID 16517654. [10] Dawson, R. Ben (1985). Data for biochemical research (3rd ed.). Oxford: Clarendon Press. p. 122. ISBN 0-19-855358-7. [11] 曾百由 編著 “數位訊號控制器原理與應用”，洪有圖書開發股份有限公司，九十三年十一月 [12] 庭田企業有限公司，網址：http://www.dichroic.com.tw/cn/index.htm。 [13] 日亞化學株式會社，網址：http://www.led-shop.com.tw/index.htm。

  43. The End

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