Abstract

Optical tweezers are widely used for noninvasive and precise micromanipulation of living cells to understand biological processes. By focusing laser beams on cells, direct cell manipulation with optical tweezers can achieve high precision and flexibility. However, direct exposure to the laser beam can lead to negative effects on the cells. These phenomena are also known as photobleaching and photodamage. In this study, we proposed a new indirect cell micromanipulation approach combined with a robot-aided holographic optical tweezer system and 3D nano-printed microtool. The microtool was designed with a V-shaped head and an optical handle part. The V-shaped head can push and trap different sizes of cells as the microtool moves forward by optical trapping of the handle part. In this way, cell exposure to the laser beam can be effectively reduced. The microtool was fabricated with a laser direct writing system by two-photon photopolymerization. A control strategy combined with an imaging processing algorithm was introduced for automated manipulation of the microtool and cells. Experiments were performed to verify the effectiveness of our approach. First, automated microtool transportation and rotation were demonstrated with high precision. Second, indirect optical transportations of cells, with and without an obstacle, were performed to demonstrate the effectiveness of the proposed approach. Third, experiments of fluorescent cell manipulation were performed to confirm that, indicated by the photobleaching effect, indirect manipulation with the microtool could induce less laser exposure compared with direct optical manipulation. The proposed method could be useful in complex biomedical applications where precise cell manipulation and less laser exposure are required.

Highlights

  • The invention of optical tweezers has allowed scientists to develop an unprecedented manipulative ability at the single-cell level [1,2,3]

  • Focused laser beams, which can trap biological cells, cell organelles, and DNA bundles in a confined space [4,5,6], are opening up unexplored research areas that focus on single cells

  • We propose a new approach for precise and safe transportation of biological cells with a robot-aided holographic optical tweezer (HOT) manipulation system

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Summary

Introduction

The invention of optical tweezers has allowed scientists to develop an unprecedented manipulative ability at the single-cell level [1,2,3]. Direct manipulation by the high focused laser beams may cause photobleaching and photodamage to the cells The photobleaching makes it difficult to precisely visualize the spatio-temporal distribution of interesting proteins in the cells by combining fluorescent probes and optical tweezers. With the advancement of the direct laser writing technique, designable microscale mechanisms with 3D nano features were printed using the two-photon photopolymerization (2PP) method and trapped as microtools for indirect optical manipulation [36]. Experiments on yeast cells, which were stained with a fluorescent dye, demonstrate that indirect optical manipulation with the proposed microtool can effectively reduce the photobleaching effect. This finding implies that laser exposure on the cells is reduced.

Microtool
Designed
Fabrication of Microtool
Experimental Setup
Control
Results
Movement of Microtool in Optical Traps and Fluid Flows
Deviations
Translation
Experiments
Rotation of Microtool
Experiments of of rotation
Transport
Transport of Cells with an Obstacle
Reduced Photobleaching
Discussion
Full Text
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