The gluten matrix in bread dough develops through the mixing process, and its microstructure is known to greatly affect the quality of the end product. In this study, a novel method to quantify the gluten matrix was developed by applying image analysis methods used in the area of bone histomorphometry to fluorescence images of dough. Acquisition of clear images of the gluten matrix and the incorporated starch has been made possible by a novel fluorescence visualization method using acid magenta and the blue fluorescent filter. The images with high contrast between gluten, starch, and other constituents allowed accurate binarization between gluten and background. Bread dough at four mixing stages was made to observe gluten development. Three parameters were extracted from the gluten area: gluten thickness, total length of gluten per area, and average length of gluten. At the final (optimum) mixing stage, gluten thickness and average length of gluten showed minimum values, while the total length of gluten showed the maximum value. This showed that at the final mixing stage, gluten strands become thin and highly branched. In addition, thin membrane-like structures were observed at the over-mixing stage, indicating the breakdown of gluten structure. Further application of this quantification method to flours with different gluten content would enable comprehensive understanding of the gluten formation in dough.
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AACC International. Approved Methods of Analysis, 11th Ed. Method 54-40.02 Mixograph Method. Approved November 3, 1999. AACC International, St. Paul, MN, U.S.A. doi:10.1094/AACCIntMethod-54-40.02
Adams, R., & Bischof, L. (1994). Seeded Region Growing. Ieee Transactions on Pattern Analysis and Machine Intelligence, 16(6), 641–647.
Autio, K., & Laurikainen, T. (1997). Relationships between flour/dough microstructure and dough handling and baking properties. Trends in Food Science & Technology, 8(6), 181–185.
Autio, K., & Salmenkallio-Marttila, M. (2001). Light microscopic investigations of cereal grains, doughs and breads. Lebensmittel-Wissenschaft Und-Technologie-Food Science and Technology, 34(1), 18–22.
Autio, K., Parkkonen, T., & Fabritius, M. (1997). Observing structural differences in wheat and rye breads. Cereal Foods World, 42(8), 702–705.
Bechtel, D. B., Pomeranz, Y., & Francisco, A. D. (1978). Breadmaking studied by light and transmission electron-microscopy. Cereal Chemistry, 55(3), 392–401.
Durrenberger, M. B., Handschin, S., Conde-Petit, B., & Escher, F. (2001). Visualization of food structure by confocal laser scanning microscopy (CLSM). Lebensmittel-Wissenschaft Und-Technologie-Food Science and Technology, 34(1), 11–17.
Fulcher, R. G., Irving, D. W., & De Francisco, A. (1989). Fluorescence microscopy: application in food analysis. In L. Munck (Ed.), Fluorescence analysis in foods (pp. 59–109). U.K: Longman Scientific and Technical, Harlow.
Gonzalez, R. C., & Woods, R. E. (2008). Digital image processing (3rd ed.). New Jersey: Pearson Education, Inc.
Hoseney, R. C. (1985). Mixing phenomenon. Cereal Foods World, 30(7).
Kokawa, M., Fujita, K., Sugiyama, J., Tsuta, M., Shibata, M., Araki, T., & Nabetani, H. (2012). Quantification of the distributions of gluten, starch and air bubbles in dough at different mixing stages by fluorescence fingerprint imaging. Journal of Cereal Science, 55(1), 15–21.
Lee, L., Ng, P. K. W., Whallon, J. H., & Steffe, J. F. (2001). Relationship between rheological properties and microstructural characteristics of nondeveloped, partially developed, and developed doughs. Cereal Chemistry, 78(4), 447–452.
Li, W., Dobraszczyk, B. J., & Wilde, P. J. (2004). Surface properties and locations of gluten proteins and lipids revealed using confocal scanning laser microscopy in bread dough. Journal of Cereal Science, 39(3), 403–411.
Lynch, E. J., Dal Bello, F., Sheehan, E. M., Cashman, K. D., & Arendt, E. K. (2009). Fundamental studies on the reduction of salt on dough and bread characteristics. Food Research International, 42(7), 885–891.
Maeda, T., Kimura, S., Araki, T., Ikeda, G., Takeya, K., & Sagara, Y. (2009). The effects of mixing stage and fermentation time on the quantity of flavor compounds and sensory intensity of flavor in white bread. Food Science and Technology Research, 15(2), 117–126.
Maeda, T., Kokawa, M., Miura, M., Araki, T., Yamada, M., Takeya, K., & Sagara, Y. (2013). Development of a novel staining procedure for visualizing the gluten–starch matrix in bread dough and cereal products. Cereal Chemistry, 90(3), 175–180.
Merz, W. A., & Schenk, R. K. (1970). Quantitative structural analysis of human cancellous bone. Acta Anatomica, 75(1), 54–66.
Nevatia, R. (1982). Machine Perception Prentice-Hall, Inc.
Paredeslopez, O., & Bushuk, W. (1983). Development and “undevelopment” of wheat dough by mixing. Cereal Chemistry, 60(1), 24–27.
Parfitt, A. M., Drezner, M. K., Glorieux, F. H., Kanis, J. A., Malluche, H., Meunier, P. J., Ott, S. M., & Recker, R. R. (1987). Bone histomorphometry - standardization of nomenclature, symbols, and units. Journal of Bone and Mineral Research, 2(6), 595–610.
Parkkonen, T., Heinonen, R., & Autio, K. (1997). A new method for determining the area of cell walls in rye doughs based on fluorescence microscopy and computer-assisted image analysis. Food Science and Technology-Lebensmittel-Wissenschaft & Technologie, 30(7), 743–747.
Peighambardoust, S. H., van der Goot, A. J., van Vliet, T., Hamer, R. J., & Boom, R. M. (2006). Microstructure formation and rheological behaviour of dough under simple shear flow. Journal of Cereal Science, 43(2), 183–197.
Peighambardoust, S. H., Dadpour, M. R., & Dokouhaki, M. (2010). Application of epifluorescence light microscopy (EFLM) to study the microstructure of wheat dough: a comparison with confocal scanning laser microscopy (CSLM) technique. Journal of Cereal Science, 51(1), 21–27.
Schluentz, E. J., Steffe, J. F., & Ng, P. K. W. (2000). Rheology and microstructure of wheat dough developed with controlled deformation. Journal of Texture Studies, 31(1), 41–54.
Simmonds, D. H. (1972). Wheat-grain morphology and its relationship to dough structure. Cereal Chemistry, 49(3), 324–335.
Upadhyay, R., Ghosal, D., & Mehra, A. (2012). Characterization of bread dough: Rheological properties and microstructure. Journal of Food Engineering, 109(1), 104–113.
Wieser, H. (2007). Chemistry of gluten proteins. Food Microbiology, 24(2), 115–119.
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Maeda, T., Kokawa, M., Nango, N. et al. Development of a Quantification Method of the Gluten Matrix in Bread Dough by Fluorescence Microscopy and Image Analysis. Food Bioprocess Technol 8, 1349–1354 (2015). https://doi.org/10.1007/s11947-015-1497-9
- Acid magenta
- Gluten skeleton
- Computer vision
- Bone histomorphometry