Lasne David, Blab Gerhard A, Berciaud Stéphane, Heine Martin, Groc Laurent, Choquet Daniel, Cognet Laurent, Lounis Brahim
Centre de Physique Moléculaire Optique et Hertzienne, CNRS (UMR 5798) and Université Bordeaux 1, Talence Cedex, France.
Biophys J. 2006 Dec 15;91(12):4598-604. doi: 10.1529/biophysj.106.089771. Epub 2006 Sep 22.
Tracking individual nano-objects in live cells during arbitrary long times is a ubiquitous need in modern biology. We present here a method for tracking individual 5-nm gold nanoparticles on live cells. It relies on the photothermal effect and the detection of the Laser Induced Scattering around a NanoAbsorber (LISNA). The key point for recording trajectories at video rate is the use of a triangulation procedure. The effectiveness of the method is tested against single fluorescent molecule tracking in live COS7 cells on subsecond timescales. We further demonstrate recordings for several minutes of AMPA receptors trajectories on the plasma membrane of live neurons. Single Nanoparticle Photothermal Tracking has the unique potential to record arbitrary long trajectory of membrane proteins using nonfluorescent nanometer-sized labels.
在现代生物学中，长时间追踪活细胞中的单个纳米物体是一种普遍需求。我们在此展示一种用于追踪活细胞上单个5纳米金纳米颗粒的方法。它依赖于光热效应以及围绕纳米吸收体的激光诱导散射（LISNA）检测。以视频速率记录轨迹的关键在于使用三角测量程序。该方法的有效性在亚秒时间尺度上针对活COS7细胞中的单个荧光分子追踪进行了测试。我们进一步展示了在活神经元质膜上记录AMPA受体轨迹长达数分钟的情况。单纳米颗粒光热追踪具有使用非荧光纳米尺寸标记记录膜蛋白任意长轨迹的独特潜力。
