Ludwig-Maximilians-University Munich, Department of Chemistry & Center for NanoScience, Butenandtstrasse 11, Gerhard-Ertl-Gebäude, 81377 Munich, Germany.
Nanomedicine (Lond). 2013 Nov;8(11):1815-28. doi: 10.2217/nnm.12.178. Epub 2013 Feb 5.
This study examines the absolute quantification of particle uptake into cells.
We developed a novel method to analyze stacks of confocal fluorescence images of single cells interacting with nano-and micro-particles. Particle_in_Cell-3D is a freely available ImageJ macro. During the image analysis routine, single cells are reconstructed in 3D and split into two volumes - intracellular and the membrane region. Next, particles are localized and color-coded accordingly. The mean intensity of single particles, measured in calibration experiments, is used to determine the absolute number of particles.
Particle_in_Cell-3D was successfully applied to measure the uptake of 80-nm mesoporous silica nanoparticles into HeLa cells. Furthermore, it was used to quantify the absolute number of 100-nm polystyrene nanoparticles forming agglomerates of up to five particles; the accuracy of these results was confirmed by super-resolution, stimulated emission depletion microscopy.
Particle_in_Cell-3D is a fast and accurate method that allows the quantification of particle uptake into cells.
本研究旨在对细胞内颗粒摄取进行绝对定量分析。
我们开发了一种新方法，用于分析与纳米和微颗粒相互作用的单个细胞的共聚焦荧光图像堆栈。Particle_in_Cell-3D 是一个免费的 ImageJ 宏。在图像分析过程中，单细胞在 3D 中重建，并分为两个区域——细胞内区和细胞膜区。然后，对颗粒进行定位并相应地进行颜色编码。在标定实验中测量的单个颗粒的平均强度用于确定颗粒的绝对数量。
Particle_in_Cell-3D 成功地应用于测量 HeLa 细胞对 80nm 介孔硅纳米颗粒的摄取。此外，它还用于定量 100nm 聚苯乙烯纳米颗粒形成多达五个颗粒聚集体的绝对数量；通过超分辨率、受激发射损耗显微镜确认了这些结果的准确性。
Particle_in_Cell-3D 是一种快速准确的方法，可用于定量细胞内颗粒摄取。
