Abstract
Laser based cell manipulation has proven to be a versatile tool in biomedical applications. In this context, combining weakly focused laser pulses and nanostructures, e.g. gold nanoparticles, promises to be useful for high throughput cell manipulation, such as transfection and photothermal therapy. Interactions between laser pulses and gold nanoparticles are well understood. However, it is still necessary to study cell behavior in gold nanoparticle mediated laser manipulation. While parameters like cell viability or perforation efficiency are commonly addressed, the influence of the manipulation process on other essential cell parameters is not sufficiently investigated yet. Thus, we set out to study four relevant cell properties: cell volume and area, ion exchange and cytoskeleton structure after gold nanoparticle based laser manipulation. For this, we designed a multimodal imaging and manipulation setup. 200 nm gold nanoparticles were attached unspecifically to canine cells and irradiated by weakly focused 850 ps laser pulses. Volume and area change in the first minute post laser manipulation was monitored using digital holography. Calcium imaging and cells expressing a marker for filamentous actin (F-actin) served to analyze the ion exchange and the cytoskeleton, respectively. High radiant exposures led to cells exhibiting a tendency to shrink in volume and area, possibly due to outflow of cytoplasm. An intracellular raise in calcium was observed and accompanied by an intercellular calcium wave. This multimodal approach enabled for the first time a comprehensive analysis of the cell behavior in gold nanoparticle mediated cell manipulation. Additionally, this work can pave the way for a better understanding and the evaluation of new applications in the context of cell transfection or photothermal therapy.
Highlights
The emerging field of nanomedicine aims to diagnose and treat various diseases with nanostructures from a hundred down to a few nanometers [1]
We focus on laser and particle parameters similar to those used in gold nanoparticle mediated laser transfection, because possible particle interactions, cell viability, and dye uptake were already analyzed in our previous publications [6, 9, 14, 15]
We examined four essential cell parameters after gold nanoparticle mediated laser manipulation: cell volume and area, filamentous actin (F-actin) cytoskeleton structure, and ion concentration
Summary
The emerging field of nanomedicine aims to diagnose and treat various diseases with nanostructures from a hundred down to a few nanometers [1]. Many applications in nanomedicine utilize gold nanoparticles. Many recent studies focus on the use of lasers as light sources and gold nanoparticles to manipulate cells [4,5,6,7]. Huang et al demonstrated applications of this configuration for plasmonic photothermal therapy [4, 5]. In this approach, gold nanoparticles served as photothermal contrast agents and their heating was used to eliminate cell carcinoma. Lukianova-Hleb employed so called plasmonic nanobubbles generated around gold nanoparticle clusters trough irradiation with short laser pulses to eliminate cells in a monolayer [7, 8]. Our group uses gold nanoparticles irradiated by picosecond laser pulses for high-throughput delivery of antisense molecules or proteins in a technique called gold nanoparticle mediated laser transfection [6, 9]
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