Displacement Detection Decoupling in Counter-Propagating Dual-Beams Optical Tweezers with Large-Sized Particle

Xunmin Zhu, Nan Li, Jianyu Yang, Xingfan Chen and Huizhu Hu

As a kind of ultra-sensitive acceleration sensing platform, optical tweezers show a minimum measurable value inversely proportional to the square of the diameter of the levitated spherical particle. However, with increasing diameter, the coupling of the displacement measurement between the axes becomes noticeable. This paper analyzes the source of coupling in a forward-scattering far-field detection regime and proposes a novel method of suppression. We theoretically and experimentally demonstrated that when three variable irises are added into the detection optics without changing other parts of optical structures, the decoupling of triaxial displacement signals mixed with each other show significant improvement. A coupling detection ratio reduction of 49.1 dB and 22.9 dB was realized in radial and axial directions, respectively, which is principally in accord with the simulations. This low-cost and robust approach makes it possible to accurately measure three-dimensional mechanical quantities simultaneously and may be helpful to actively cool the particle motion in optical tweezers even to the quantum ground state in the future.

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Fabrication of Multimode-Single Mode Polymer Fiber Tweezers for Single Cell Trapping and Identification with Improved Performance

Sandra M. Rodrigues, Joana S. Paiva, Rita S. R. Ribeiro, Olivier Soppera, João P. S. Cunha and Pedro A. S. Jorge

Optical fiber tweezers have been gaining prominence in several applications in Biology and Medicine. Due to their outstanding focusing abilities, they are able to trap and manipulate microparticles, including cells, needing any physical contact and with a low degree of invasiveness to the trapped cell. Recently, we proposed a fiber tweezer configuration based on a polymeric micro-lens on the top of a single mode fiber, obtained by a self-guided photopolymerization process. This configuration is able to both trap and identify the target through the analysis of short-term portions of the back-scattered signal. In this paper, we propose a variant of this fabrication method, capable of producing more robust fiber tips, which produce stronger trapping effects on targets by as much as two to ten fold. These novel lenses maintain the capability of distinguish the different classes of trapped particles based on the back-scattered signal. This novel fabrication method consists in the introduction of a multi mode fiber section on the tip of a single mode (SM) fiber. A detailed description of how relevant fabrication parameters such as the length of the multi mode section and the photopolymerization laser power can be tuned for different purposes (e.g., microparticles trapping only, simultaneous trapping and sensing) is also provided, based on both experimental and theoretical evidences.

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Microfluidic Cultivation and Laser Tweezers Raman Spectroscopy of E. coli under Antibiotic Stress

Zdeněk Pilát, Silvie Bernatová, Jan Ježek, Johanna Kirchhoff, Astrid Tannert, Ute Neugebauer, Ota Samek and Pavel Zemánek

Analyzing the cells in various body fluids can greatly deepen the understanding of the mechanisms governing the cellular physiology. Due to the variability of physiological and metabolic states, it is important to be able to perform such studies on individual cells. Therefore, we developed an optofluidic system in which we precisely manipulated and monitored individual cells of Escherichia coli. We tested optical micromanipulation in a microfluidic chamber chip by transferring individual bacteria into the chambers. We then subjected the cells in the chambers to antibiotic cefotaxime and we observed the changes by using time-lapse microscopy. Separately, we used laser tweezers Raman spectroscopy (LTRS) in a different micro-chamber chip to manipulate and analyze individual cefotaxime-treated E. coli cells. Additionally, we performed conventional Raman micro-spectroscopic measurements of E. coli cells in a micro-chamber. We found observable changes in the cellular morphology (cell elongation) and in Raman spectra, which were consistent with other recently published observations. The principal component analysis (PCA) of Raman data distinguished between the cefotaxime treated cells and control. We tested the capabilities of the optofluidic system and found it to be a reliable and versatile solution for this class of microbiological experiments.

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Single Particle Differentiation through 2D Optical Fiber Trapping and Back-Scattered Signal Statistical Analysis: An Exploratory Approach

Joana S. Paiva, Rita S. R. Ribeiro, João P. S. Cunha, Carla C. Rosa and Pedro A. S. Jorge

Recent trends on microbiology point out the urge to develop optical micro-tools with multifunctionalities such as simultaneous manipulation and sensing. Considering that miniaturization has been recognized as one of the most important paradigms of emerging sensing biotechnologies, optical fiber tools, including Optical Fiber Tweezers (OFTs), are suitable candidates for developing multifunctional small sensors for Medicine and Biology. OFTs are flexible and versatile optotools based on fibers with one extremity patterned to form a micro-lens. These are able to focus laser beams and exert forces onto microparticles strong enough (piconewtons) to trap and manipulate them. In this paper, through an exploratory analysis of a 45 features set, including time and frequency-domain parameters of the back-scattered signal of particles trapped by a polymeric lens, we created a novel single feature able to differentiate synthetic particles (PMMA and Polystyrene) from living yeasts cells. This single statistical feature can be useful for the development of label-free hybrid optical fiber sensors with applications in infectious diseases detection or cells sorting. It can also contribute, by revealing the most significant information that can be extracted from the scattered signal, to the development of a simpler method for particles characterization (in terms of composition, heterogeneity degree) than existent technologies.

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Effects of Infrared Optical Trapping on Saccharomyces cerevisiae in a Microfluidic System

Zdeněk Pilát, Alexandr Jonáš, Jan Ježek and Pavel Zemánek

Baker’s yeast (Saccharomyces cerevisiae) represents a very popular single-celled eukaryotic model organism which has been studied extensively by various methods and whose genome has been completely sequenced. It was also among the first living organisms that were manipulated by optical tweezers and it is currently a frequent subject of optical micromanipulation experiments. We built a microfluidic system for optical trapping experiments with individual cells and used it for the assessment of cell tolerance to phototoxic stress. Using optical tweezers with the wavelength of 1064 nm, we trapped individual Saccharomyces cerevisiae cells for 15 min and, subsequently, observed their stress response in specially designed microfluidic chambers over time periods of several hours by time-lapse video-microscopy. We determined the time between successive bud formations after the exposure to the trapping light, took account of damaged cells, and calculated the population doubling period and cell areas for increasing trapping power at a constant trapping time. Our approach represents an attractive, versatile microfluidic platform for quantitative optical trapping experiments with living cells. We demonstrate its application potential by assessing the limits for safe, non-invasive optical trapping of Saccharomyces cerevisiae with infrared laser light.

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Probing the Kinetic Anabolism of Poly-Beta-Hydroxybutyrate in Cupriavidus necator H16 Using Single-Cell Raman Spectroscopy

Zhanhua Tao, Lixin Peng, Pengfei Zhang, Yong-Qing Li and Guiwen Wang

Poly-beta-hydroxybutyrate (PHB) can be formed in large amounts in Cupriavidus necator and is important for the industrial production of biodegradable plastics. In this investigation, laser tweezers Raman spectroscopy (LTRS) was used to characterize dynamic changes in PHB content—as well as in the contents of other common biomolecule—in C. necator during batch growth at both the population and single-cell levels. PHB accumulation began in the early stages of bacterial growth, and the maximum PHB production rate occurred in the early and middle exponential phases. The active biosynthesis of DNA, RNA, and proteins occurred in the lag and early exponential phases, whereas the levels of these molecules decreased continuously during the remaining fermentation process until the minimum values were reached. The PHB content inside single cells was relatively homogenous in the middle stage of fermentation; during the late growth stage, the variation in PHB levels between cells increased. In addition, bacterial cells in various growth phases could be clearly discriminated when principle component analysis was performed on the spectral data. These results suggest that LTRS is a valuable single-cell analysis tool that can provide more comprehensive information about the physiological state of a growing microbial population.

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Raman Spectroscopy of Optically Trapped Single Biological Micro-Particles

Brandon Redding, Mark Schwab and Yong-le Pan

The combination of optical trapping with Raman spectroscopy provides a powerful method for the study, characterization, and identification of biological micro-particles. In essence, optical trapping helps to overcome the limitation imposed by the relative inefficiency of the Raman scattering process. This allows Raman spectroscopy to be applied to individual biological particles in air and in liquid, providing the potential for particle identification with high specificity, longitudinal studies of changes in particle composition, and characterization of the heterogeneity of individual particles in a population. In this review, we introduce the techniques used to integrate Raman spectroscopy with optical trapping in order to study individual biological particles in liquid and air. We then provide an overview of some of the most promising applications of this technique, highlighting the unique types of measurements enabled by the combination of Raman spectroscopy with optical trapping. Finally, we present a brief discussion of future research directions in the field.

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Spectroscopy, Manipulation and Trapping of Neutral Atoms, Molecules, and Other Particles Using Optical Nanofibers: A Review

Michael J. Morrissey, Kieran Deasy, Mary Frawley, Ravi Kumar, Eugen Prel, Laura Russell, Viet Giang Truong and Síle Nic Chormaic

The use of tapered optical fibers, i.e., optical nanofibers, for spectroscopy and the detection of small numbers of particles, such as neutral atoms or molecules, has been gaining interest in recent years. In this review, we briefly introduce the optical nanofiber, its fabrication, and optical mode propagation within. We discuss recent progress on the integration of optical nanofibers into laser-cooled atom and vapor systems, paying particular attention to spectroscopy, cold atom cloud characterization, and optical trapping schemes. Next, a natural extension of this work to molecules is introduced. Finally, we consider several alternatives to optical nanofibers that display some advantages for specific applications.

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Trapping and Propelling Microparticles at Long Range by Using an Entirely Stripped and Slightly Tapered No-Core Optical Fiber

Fang-Wen Sheu and Yen-Si Huang

A stripped no-core optical fiber with a 125 µm diameter was transformed into a symmetric and unbroken optical fiber that tapers slightly to a 45-µm-diameter waist. The laser light can be easily launched into the no-core optical fiber. The enhanced evanescent wave of the slightly tapered no-core optical fiber can attract nearby 5-µm-diameter polystyrene microparticles onto the surface of the tapered multimode optical fiber within fast flowing fluid and propel the trapped particles in the direction of the light propagation to longer delivery range than is possible using a slightly tapered telecom single-mode optical fiber.

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On-Chip Cellomics Assay Enabling Algebraic and Geometric Understanding of Epigenetic Information in Cellular Networks of Living Systems. 1. Temporal Aspects of Epigenetic Information in Bacteria

Kenji Yasuda

A series of studies aimed at developing methods and systems of analyzing epigenetic information in cells and in cell networks, as well as that of genetic information, was examined to expand our understanding of how living systems are determined. Because cells are minimum units reflecting epigenetic information, which is considered to map the history of a parallel-processing recurrent network of biochemical reactions, their behaviors cannot be explained by considering only conventional DNA information-processing events. The role of epigenetic information on cells, which complements their genetic information, was inferred by comparing predictions from genetic information with cell behaviour observed under conditions chosen to reveal adaptation processes, population effects and community effects. A system of analyzing epigenetic information was developed starting from the twin complementary viewpoints of cell regulation as an “algebraic” system (emphasis on temporal aspects) and as a “geometric” system (emphasis on spatial aspects). Exploiting the combination of latest microfabrication technology and measurement technologies, which we call on-chip cellomics assay, we can control and re-construct the environments and interaction of cells from “algebraic” and “geometric” viewpoints. In this review, temporal viewpoint of epigenetic information, a part of the series of single-cell-based “algebraic” and “geometric” studies of celluler systems in our research groups, are summerized and reported. The knowlege acquired from this study may lead to the use of cells that fully control practical applications like cell-based drug screening and the regeneration of organs.

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