CAJ | 학술논문

单分子荧光检测作为一种能够表征分子个体性质及行为的分析方法，有助于揭示利用传统荧光检测方法无法得到的信息，在近年来受到人们的广泛关注。利用传统光学检测设备进行单分子荧光检测时，由于受到衍射极限的限制，同时为了保证在观测体积内只有单个荧光分子，仅能采用无限稀释溶液的方法实现单分子荧光检测。虽然这种方法可以满足单分子检测的要求，但是由于大部分酶分子正常工作时底物的生理浓度都非常高，底物浓度的大幅度降低会对酶分子的反应机制等方面造成影响。零模波导作为一种新型的单分子检测器件，通过纳米微孔结构突破了光学衍射极限的限制将观测体积降至仄升量级（10－21L），使得在生理浓度范围内检测单分子荧光成为可能，在单分子荧光检测领域得到了广泛应用。因此，就零模波导的原理、制备工艺及其在单分子DNA测序、生物膜、生物大分子之间的相互作用及单分子反应动力学方面的具体应用进行综述。
단분자형광검측작위일충능구표정분자개체성질급행위적분석방법，유조우게시이용전통형광검측방법무법득도적신식，재근년래수도인문적엄범관주。이용전통광학검측설비진행단분자형광검측시，유우수도연사겁한적한제，동시위료보증재관측체적내지유단개형광분자，부능채용무한희석용액적방법실현단분자형광검측。수연저충방법가이만족단분자검측적요구，단시유우대부분매분자정상공작시저물적생리농도도비상고，저물농도적대폭도강저회대매분자적반응궤제등방면조성영향。령모파도작위일충신형적단분자검측기건，통과납미미공결구돌파료광학연사겁한적한제장관측체적강지측승량급（10－21L），사득재생리농도범위내검측단분자형광성위가능，재단분자형광검측영역득도료엄범응용。인차，취령모파도적원리、제비공예급기재단분자DNA측서、생물막、생물대분자지간적상호작용급단분자반응동역학방면적구체응용진행종술。
Techniques for single fluorescent molecules detection have garnered a great amount of interests from science community in recent years largely due to their ability to reveal the individual molecular properties and behaviors which were often obscured by techniques based on ensemble average.In order to overcome diffraction-limit of optical observation, fluorescent molecules must be diluted to a very low concentration-way below normal physiological concentration.However, this may adversely impact the kinetics for the chemical or biochemical reaction under study.Zero-mode waveguides ( ZMWs ) , as a new kind of single fluorescent molecule detection device, effectively reduce the observation volume to the level of zeptoliter ( 10-21 L) , which is much lower than optical diffraction-limited volume, by utilizing of a nanoaperture structure.The zeptoliter-observation volume provides an environment where a single fluorescent molecule can be observed at normal physiological concentration.The ZMWs have been extensively used in the field of single fluorescent molecule detection due to its outstanding performance than diffraction-limited optics.Therefore we provided an overview of ZMWs, including discussions on its principle, fabrication techniques, and applications in DNA sequencing, biomembrane structure elucidation, biomacromolecule interaction studies and kinetics study of single-molecule reaction.