De Luca Elisa, Pedone Deborah, Scarsi Anna, Marotta Roberto, Catalano Federico, Debellis Doriana, Cursi Lorenzo, Grimaldi Benedetto, Moglianetti Mauro, Pompa Pier Paolo
Nanobiointeractions&Nanodiagnostics Istituto Italiano di Tecnologia via Morego 30 16163 Genova Italy.
Institute of Nanotechnology (NANOTEC) National Research Council Via Monteroni 73100 Lecce Italy.
Small Sci. 2024 Jun 9;4(9):2400085. doi: 10.1002/smsc.202400085. eCollection 2024 Sep.
The highly efficient peroxidase-like activity of platinum nanozymes (3-20 nm size) is exploited within the complex cellular environment to catalyze the oxidation of the DAB substrate, producing an electron-dense signal around the nanozyme surface, upon osmium staining. It is proved that such nanozyme amplification can achieve a catalytic signal enhancement up to 10-fold, enabling the quick detection of the Pt particles (even of 3 nm size) by transmission electron microscopy (TEM) also at low magnification and across wide fields of view in the intricate intracellular milieu. The developed procedure is ideally suited to overcome standard amplification strategies currently used in TEM analysis, such as gold or silver enhancements. Furthermore, the wide versatility of the Pt-nanozyme probes in TEM imaging is demonstrated in immuno-EM and protein trafficking studies, showing their potential to track the subcellular localization of target biomolecules at both low and high magnifications. These results suggest that the use of nanozymes might represent a paradigm shift in the conventional amplification systems currently employed in electron microscopy for cellular analyses, offering enhanced imaging capabilities.
铂纳米酶(尺寸为3 - 20纳米)具有高效的类过氧化物酶活性,在复杂的细胞环境中可利用其催化DAB底物的氧化反应,经锇染色后,在纳米酶表面周围产生电子致密信号。事实证明,这种纳米酶放大作用可实现高达10倍的催化信号增强,使得通过透射电子显微镜(TEM)在低放大倍数下也能快速检测到铂颗粒(甚至3纳米大小的颗粒),且能在复杂的细胞内环境中实现宽视野观察。所开发的方法非常适合克服目前TEM分析中使用的标准放大策略,如金或银增强。此外,铂纳米酶探针在免疫电镜和蛋白质转运研究中的TEM成像中具有广泛的通用性,显示出它们在低倍和高倍放大下追踪目标生物分子亚细胞定位的潜力。这些结果表明,纳米酶的使用可能代表了目前电子显微镜用于细胞分析的传统放大系统的范式转变,提供了增强的成像能力。