Abstract
Milk intended for human consumption is subjected to technological treatments to ensure its safety and storage stability. These treatments deeply modify some of its structural and nutritional characteristics. Principal modifications involve proteins that partly adsorb onto the membrane of milk fat globules upon homogenization or whey proteins that undergo denaturation and aggregation during thermal treatments. Transmission electron microscopy is a powerful approach to investigate milk ultrastructure, due to its high-resolution power. Immunogold labeling of β-lactoglobulin and β-casein proteins represents a sophisticated approach to examine their structure and localization following technological processes such as homogenization and UHT treatment. However, liquid milk is a very challenging matrix because of its complex multiphasic nature. To preserve both ultrastructure and antigenicity, and to obtain an efficient labelling in liquid milk samples, some precautions shall be adopted in fixation, embedding, and labeling steps as here reported.
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References
Fox PF, Kelly AL (2006) Chemistry and biochemistry of milk constituents. Food Biochem Food Process 2:442–464
Marangoni F, Pellegrino L, Verduci E, Ghiselli A, Bernabei R, Calvani R et al (2019) Cow’s milk consumption and health: a health professional’s guide. J Am Coll Nut 38:197–208
Ménard O, Ahmad S, Rousseau F, Briard-Bion V, Gaucheron F, Lopez C (2010) Buffalo vs. cow milk fat globules: size distribution, zeta-potential, compositions in total fatty acids and in polar lipids from the milk fat globule membrane. Food Chem 120:544–551
Pellegrino L, D’Incecco P (2022) Impact of processing on nutritional quality of milk proteins. In: McSweeney PLH, McNamara JP (eds) Encyclopedia of dairy science, vol 5. Elsevier, Academic, Amsterdam, pp 859–866
Michalski MC, Januel C (2006) Does homogenization affect the human health properties of cow’s milk? Trends Food Sci Technol 17:423–437
D’Incecco P, Rosi V, Cabassi G, Hogenboom JA, Pellegrino L (2018) Microfiltration and ultra-high-pressure homogenization for extending the shelf-storage stability of UHT milk. Food Res Int 107:477–485
Lee SJ, Sherbon JW (2002) Chemical changes in bovine milk fat globule membrane caused by heat treatment and homogenization of whole milk. J Dairy Res 69:555–567
Newman GR, Hobot JA (2001) Resin microscopy and on-section immuno-cytochemistry. Springer Lab Manuals, Springer, Berlin
Hillbrick GC, McMahon DJ, McManus WR (1999) Microstructure of indirectly and directly heated ultra-high-temperature (UHT) processed milk examined using transmission electron microscopy and immunogold labelling. LWT Food Sci Technol 32:486–494
D’Incecco P, Ong L, Pellegrino L, Faoro F, Barbiroli A, Gras S (2018) Effect of temperature on the microstructure of fat globules and the immunoglobulin-mediated interactions between fat and bacteria in natural raw milk creaming. J Dairy Sci 101:2984–2997
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D’Incecco, P. (2023). Immunogold Labeling of Milk Proteins at Transmission Electron Microscopy. In: Pellicciari, C., Biggiogera, M., Malatesta, M. (eds) Histochemistry of Single Molecules. Methods in Molecular Biology, vol 2566. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-2675-7_12
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DOI: https://doi.org/10.1007/978-1-0716-2675-7_12
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