Introduction

Abstract

Real-time high-throughput identification, high content screening, characterisation and processing of reflective micro and nano structures as well as (semi-)transparent phase objects like biological specimens are of significant interest to a variety of areas ranging from cell biology and medicine to lithography. In this thesis, the two optical techniques, namely digital holographic microscopy (DHM) and holographic optical tweezers (HOTs) are realised in one integrated setup that permits to minimal-invasively image, manipulate, control, sense, track and identify micro and nano scaled specimens or even fabricate them with direct laser writing in three dimensions. The setup allows to operate both methods simultaneously additionally to conventional bright field, confocal laser scanning (LSM) and fluorescence microscopy. This enables e.g. time lapse experiments in microfluidic environments with stably arranged or distanced cells respectively. Furthermore the combination of both holographic methods allows to examine and monitor e.g. cell deformations, their volume or reactions to certain artificially manipulated surface types as well as apoptosis and the influence radius of released messengers. For a better signal to noise ratio, different low coherent light sources including supercontinuum light sources are investigated. The system setup and its functionality are characterised in this work. Holographic optical tweezers not only open up the possibility for generating multiple dynamic traps for micro and nano particles, but also the opportunity to exert optical torque with special complex electromagnetic fields like Bessel beams, which can facilitate the movement and rotation of particles. Further non diffracting beams are investigated for utilisation in the holographic optical tweezer (HOT) system for self-assembling of micro particles which comprises Mathieu, Laguerre and Airy beams. New opportunities that arise from the complex methods are investigated also for direct laser writing of nano structures based on two photon polymerisation.