流式细胞仪检测微细胶黏物中尼罗红染料选择性吸附的研究

Study on Selective Adsorption of Nile Red in Micro-stickies Detection by Flow Cytometry

关键词:  微细胶黏物  流式细胞仪  Nile Red染料  吸附

Key Words:micro-stickies  flow cytometry  Nile Red dye  adsorption

摘要:采用尼罗红（Nile Red）染料分别对旧报纸（ONP）浆和旧瓦楞箱纸板（OCC）浆的白水、苯-醇抽提前后漂白硫酸盐针叶木浆（漂白浆）的白水、纤维素酶处理前后的漂白浆白水进行染色，随后用流式细胞仪检测分析。结果表明，Nile Red染料可吸附废纸浆白水中疏水性物质，在检测过程中激发荧光产生标记。但用自来水（模拟造纸用水）也可被激发荧光，表明废纸浆中可被检测的颗粒不全是微细胶黏物；使用苯-醇抽提漂白浆以排除抽出物的干扰，抽提后其白水中颗粒浓度为269个/μL，颗粒平均粒径为3.24 μm，与抽提前相比无明显变化，表明不含抽出物漂白浆的白水中仍存在可被Nile Red染料吸附的物质；纤维素酶处理后白水中颗粒浓度从246个/μL减少到61个/μL，颗粒平均粒径从3.19 μm减少为2.98 μm，细小纤维的降解使白水中能够被染料吸附的颗粒减少，表明漂白浆白水中细小纤维可被Nile Red染料吸附并影响检测结果。最后确定流式细胞仪检测废纸浆颗粒浓度结果校正系数为85%。

Abstract:In this study，Nile Red dye was used to dye white water of ONP pulp and OCC pulp，white water of bleached kraft softwood pulp extracted with benzene-alcohol，white water of bleached kraft softwood pulp before and after cellulase treatment，and then flow cytometry was used for detection and analysis. The results showed that the Nile Red dye could adsorb hydrophobic substances in the white water of waste paper pulp and excite fluorescence during the detection process. But using tap water (simulated papermaking water) could also excite fluorescence，which indicated that the detected particals in waste paper pulp were not all micro-stickies；Benzene-alcohol was used to extract the bleached kraft softwood pulp to eliminate the interference of extractants. After extraction，the concentration of particles in white water was 269/μL，and the average particle size was 3.24 μm，which showed that no significant change compared with before extracfion，indicated that there were still substances adsorbed by Nile Red dye in white water that did not contain extractants. Particles concentration in white water of bleached kraft softwood pulp before and after cellulase treatment was reduced from 246/μL to 61/μL，average particle size reduced from 3.19 μm to 2.98 μm，the degradation of fines could decrease the mount of absrbing particies by dye in the water， indicating that the fine could be adsorbed by Nile Red dye and affecting the detection results.Finally，the calibration coefficient of laboratory flow cytometry was determined to be 85%.