Versatile optical manipulation system for inspection, laser processing, and isolation of individual living cells

Abstract

Isolation of individual cells from a heterogeneous cell population is an invaluable step in the analysis of single cell properties. The demands in molecular and cellular biology as well as molecular medicine are the selection, isolation, and monitoring of single cells and cell clusters of biopsy material. Of particular interest are methods which complement a passive optical or spectroscopic selection with a variety of active single cell processing techniques such as mechanical, biochemical, or genetic manipulation prior to isolation. Sophisticated laser-based cell processing systems are available which can perform single cell processing in a contact-free and sterile manner. Until now, however, these multipurpose turnkey systems offer only basic micromanipulation and are not easily modified or upgraded, whereas laboratory situations often demand simple but versatile and adaptable solutions. We built a flexible laser micromanipulation platform combining contact-free microdissection and catapulting capabilities using a pulsed ultraviolet (337nm) laser with simultaneous generation of optical tweezing forces using a continuous wave infrared (1064nm) laser. The potential of our platform is exemplified with techniques such as local laser-induced injection of biomolecules into individual living cells, laser surgery, isolation of single cells by laser catapulting, and control of neuronal growth using optical gradient forces. Arbitrary dynamic optical force patterns can be created by fast laser scanning with acousto-optical deflectors and galvanometer mirrors, allowing multibeam contact-free micromanipulation, a prerequisite for reliable handling of material in laboratory-on-a-chip applications. All common microscopy techniques can be used simultaneously with the offered palette of micromanipulation methods. Taken together, we show that advanced optical micromanipulation systems can be designed which combine quality, cost efficiency, and adaptability.

Engineering Hydrogels as Extracellular Matrix Mimics

“…This approach offers the ability to deposit cell-encapsulating microgels at predetermined locations with multiple cell types at high throughput [107,137]. Currently, several printing techniques have been explored to generate cell-encapsulating gel droplets, such as inkjet [136,188–189], laser [190–192] and acoustic printing [193–195]. Furthermore, a scaffold-free approach has been developed to build cell encapsulating hydrogels with customized tubular structures of defined topology (both linear and branched tubular structures) (Figure 6) [187].…”

Optical tweezers for single cells

“…Florescent biomolecules have been injected into the cytosol and nucleus of Neuro-2A mouse neuroblastoma cells (Stuhrmann et al 2006).…”

Vitamin E TPGS-Emulsified Poly(Lactic-Co-Glycolic Acid) Nanoparticles for Cardiovascular Restenosis Treatment

“…In the case of subcellular laser ablation, efforts are ongoing to increase the spatial and temporal resolution of material destruction by using pulsed lasers that offer optimal combinations of pulse width, repetition rate, and energy (Vogel et al 2005). Moreover, because lasers may be used for capturing particles (eg, through an optical trap), cell stretching, and a variety of other processing modalities, efforts are beginning to emerge to incorporate laser ablation into multimodular laser processing systems, enabling microdissection and capturing capabilities in a single platform (Stuhrmann et al 2006). Similarly, because laser ablation is a multiphoton process that may be induced with the same sorts of near-infrared lasers that are typically used for multiphoton imaging, it has become attractive to develop combined systems for imaging and ablation.…”