激光共聚焦显微法测定面团面筋网络结构的影响因素  

Influencing Factors of Gluten Network Structure Analysis Using Confocal Laser Scanning Microscopy

作　　者：姜继凯 张影全[1] 郭波莉[1] 杨静洁 黄璐瑶 李明[1] 钟耕[2] JIANG JiKai;ZHANG YingQuan;GUO BoLi;YANG JingJie;HUANG LuYao;LI Ming;ZHONG Geng(Institute of Food Science and Technology,Chinese Academy of Agricultural Sciences/Comprehensive Utilization Laboratory of Cereal and Oil Processing,Ministry of Agriculture and Rural Affairs,Beijing 100193;College of Food Science Southwest University,Chongqing 400715)

机构地区：[1]中国农业科学院农产品加工研究所/农业农村部粮油加工综合利用技术集成实验室,北京100193 [2]西南大学食品科学学院,重庆400715

出　　处：《中国农业科学》2023年第14期2787-2797,共11页Scientia Agricultura Sinica

基　　金：现代农业产业技术体系建设专项(CARS-03);中国农业科学院科技创新工程(CAAS-ASTIP-2020-IFST)。

摘　　要：【目的】面团样品切片、混合染色剂浓度以及染色时间是激光共聚焦显微镜测定面团面筋网络结构的关键影响因素。本研究旨在分析这些因素对面团面筋网络结构测定结果的影响规律,进而优化出最佳的切片厚度和染色条件,为面团面筋网络结构及面制品品质高质量研究提供技术支撑。【方法】选取面筋筋力不同的两个小麦品种郑麦366(Z366)和小偃22(X22)为试验材料,磨粉制作成面团,并分别冷冻0 d和1 d;用冷冻切片机将面团分别切成10、14和20μm厚度的薄片,并配制不同浓度罗丹明B(RDB)和异硫氰酸荧光素(FITC)混合染液,分别染色1、10和20 min,分析这些因素对激光共聚焦显微法(CLSM)测定面团面筋网络结构的影响。【结果】切片厚度为10μm时,郑麦366(Z366)和小偃22(X22)面团的面筋网络结构均最清晰,且在此切片厚度下,两个品种冷冻0 d和冷冻1 d的面团面筋网络结构之间的差异均最显著(P<0.01)。染色条件研究结果表明,低浓度染色剂(0.001%+0.01%)染色时间越长,面筋蛋白网络结构呈现越清晰;高浓度染色剂(0.025%+0.25%)在染色1 min时,面筋网络结构呈现最佳,染色10 min时,面筋网络结构最差。【结论】面团切片厚度10μm,不同筋力和不同冷冻处理的面团网络结构差异最大,即面筋网络结构区分效果最明显;在此切片厚度条件下,当不考虑时间成本时,运用低浓度混合染色剂染色20 min面筋网络结构呈现效果最佳;当需要节省时间并观察淀粉颗粒细微结构时,使用高浓度染色剂染色1 min效果最好。【Objective】The thickness of sample slices,the concentration of staining solution and the staining time are the key factors influencing the results of gluten network structure analysis using confocal laser scanning microscopy(CLSM).The aim of this study was to investigate the effects of the slice thickness,the staining solution concentration and the staining time on the CLSM results of gluten network structure,and the optimal slice thickness and staining conditions for the dough structure analysis were selected,so as to provide a technical support for the studies on dough structure and/or its final products.【Method】Two wheat varieties,i.e.,Zhengmai 366(Z366)and Xiaoyan 22(X22)with different wet gluten indexes,were selected as raw materials,which were milled and made into dough,respectively.Then,the dough samples were frozen for 0 and 1 day.The dough slice thickness was set as 10,14 and 20μm,respectively.Meanwhile,two concentrations of staining solution(0.001%+0.01%,0.025%+0.25%)using Rhodamine B(RDB)and fluorescein isothiocyanate(FITC)were prepared.The staining time was set for 1,10 and 20 min,respectively.【Result】The sample slice with 10μm thickness showed the best and clearest image for both doughs of Z366 and X22.Besides,under this slice thickness,the gluten network of the dough frozen stored for 0 day showed the most obvious difference compared with that for 1 day(P<0.01).Additionally,under low-concentration staining agent(0.001%+0.01%),the clearer the results of gluten network structure were obtained with longer staining time.Oppositely,under the high-concentration staining agent(0.025%+0.25%),the slice with a staining time of 1 min showed the most rational results on the gluten network structure,whereas the result on the slice with a staining time of 10 min was the worst.【Conclusion】The gluten network structure of dough with different protein content and freezing treatments demonstrated more differences when the thickness of dough slices was 10μm,indicating the gluten network structure of

关 键 词：激光共聚焦显微法 面筋网络结构 切片厚度 混合染色剂 染色条件 

分 类 号：TS211.7[轻工技术与工程—粮食、油脂及植物蛋白工程]