Skip to main content

Development of a Quantification Method of the Gluten Matrix in Bread Dough by Fluorescence Microscopy and Image Analysis

Abstract

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.

This is a preview of subscription content, access via your institution.

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5

References

  1. 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

  2. Adams, R., & Bischof, L. (1994). Seeded Region Growing. Ieee Transactions on Pattern Analysis and Machine Intelligence, 16(6), 641–647.

    Article  Google Scholar 

  3. 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.

    Article  CAS  Google Scholar 

  4. 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.

    Article  CAS  Google Scholar 

  5. Autio, K., Parkkonen, T., & Fabritius, M. (1997). Observing structural differences in wheat and rye breads. Cereal Foods World, 42(8), 702–705.

    Google Scholar 

  6. Bechtel, D. B., Pomeranz, Y., & Francisco, A. D. (1978). Breadmaking studied by light and transmission electron-microscopy. Cereal Chemistry, 55(3), 392–401.

    Google Scholar 

  7. 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.

    Article  CAS  Google Scholar 

  8. 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.

    Google Scholar 

  9. Gonzalez, R. C., & Woods, R. E. (2008). Digital image processing (3rd ed.). New Jersey: Pearson Education, Inc.

    Google Scholar 

  10. Hoseney, R. C. (1985). Mixing phenomenon. Cereal Foods World, 30(7).

  11. 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.

    Article  CAS  Google Scholar 

  12. 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.

    Article  CAS  Google Scholar 

  13. 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.

    Article  CAS  Google Scholar 

  14. 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.

    Article  CAS  Google Scholar 

  15. 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.

    Article  CAS  Google Scholar 

  16. 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.

    Article  CAS  Google Scholar 

  17. Merz, W. A., & Schenk, R. K. (1970). Quantitative structural analysis of human cancellous bone. Acta Anatomica, 75(1), 54–66.

    Article  CAS  Google Scholar 

  18. Nevatia, R. (1982). Machine Perception Prentice-Hall, Inc.

  19. Paredeslopez, O., & Bushuk, W. (1983). Development and “undevelopment” of wheat dough by mixing. Cereal Chemistry, 60(1), 24–27.

  20. 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.

  21. 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.

    Article  CAS  Google Scholar 

  22. 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.

    Article  CAS  Google Scholar 

  23. 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.

    Article  Google Scholar 

  24. 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.

    Article  Google Scholar 

  25. Simmonds, D. H. (1972). Wheat-grain morphology and its relationship to dough structure. Cereal Chemistry, 49(3), 324–335.

    CAS  Google Scholar 

  26. Upadhyay, R., Ghosal, D., & Mehra, A. (2012). Characterization of bread dough: Rheological properties and microstructure. Journal of Food Engineering, 109(1), 104–113.

    Article  Google Scholar 

  27. Wieser, H. (2007). Chemistry of gluten proteins. Food Microbiology, 24(2), 115–119.

    Article  CAS  Google Scholar 

Download references

Author information

Affiliations

Authors

Corresponding author

Correspondence to Tatsuro Maeda.

Rights and permissions

Reprints and Permissions

About this article

Verify currency and authenticity via CrossMark

Cite this article

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

Download citation

Keywords

  • Acid magenta
  • Gluten skeleton
  • Computer vision
  • Bone histomorphometry