Abstract

Robot-assisted cell manipulation is gaining attention for its ability in providing high throughput and high precision cell manipulation for the biological industry. This paper presents a visual servo microrobotic system for cell microinjection. We investigated the automatic cell autofocus method that reduced the complexity of the system. Then, we produced an adaptive visual processing algorithm to detect the location of the cell and micropipette toward the uneven illumination problem. Fourteen microinjection experiments were conducted with zebrafish embryos. A 100% success rate was achieved either in autofocus or embryo detection, which verified the robustness of the proposed automatic cell manipulation system.

Highlights

  • Microinjecting microliters of genetic material into embryos of model animals is a standard method used for analyzing vertebrate embryonic development and the pathogenic mechanisms of human disease [1,2]

  • Position control with force signal feedback was used by Lu et al to inject zebrafish embryos, where a piezoresistive microforce sensor was used to monitor the injection process [5]

  • The injection process was as follows: 1. the petri dish containing the embryos and culture medium was placed under the microscope; 2. the embryo was autofocused by using the autofocusing algorithm; 3. the injection pipette was moved to the focus plane; 4. the adaptive image processing was used to get the location and dimension information of the embryo; M5.icromthacehitneems 2p01la6,te7,m10a4tching algorithm was used to obtain the location of the pipette tip; 6. the distance between the center of the cell and pipette tip along the x-axis and y-axis was calculated; F7i.gurtehe12inbjeschtioownsptihpeetsteucwceasssafuultodmetaetcictiaolnlyomf othveedeminbtorycoenatnedr otfhtehpeiepmetbterytoip;, and Figure 12c is the i8m. agtehoefstahmepelme bwraysodaefpteorsaituetdominatoticthienjyeoctlikosne.ction of the embryo; 9. tThaebpleip4etstehoowutsmthoeverdesoufltthseoefmthberyaou. tomatic microinjection experiments

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Summary

Introduction

Microinjecting microliters of genetic material into embryos of model animals is a standard method used for analyzing vertebrate embryonic development and the pathogenic mechanisms of human disease [1,2]. Several robotic injection prototypes for cell microinjection were reported [4,5,6,7,8,9]. Wang et al used a position control strategy to inject zebrafish embryos, in which a visual servoing method was used to detect the target position of the end-effector, and a PID (proportional-integral-derivative) position control was used for micropipette movement [4]. Xie et al employed an explicit force control method to regulate the cell injection force on zebrafish embryos [8,9]. These studies focused on the motorized injection strategy and control algorithm, even though vision feedback was adopted in every robotic prototype. The integrated interface software platform for visual servoing control was developed under the MicroTshoeftinVtiesguraaltCed++in(t6e.0rf)aecnevsioroftnwmaernetptloatefnosrumrefothrevcisoumapl asetirbvioliitnygancodnptroortlawbialsitydeavmeolonpgetdheunsodfetwr tahree MmiocdrousluofstaVndisuhaalrdCw++are(6.

Visual Servoing Algorithm for Automatic Cell Microinjection
Implementation of the Automatic Focusing Method
Adaptive Image Processing Algorithm for Automatic Cell and Pipette Detection
Detection of Injection Pipette Tip
Experiments
Conclusions
Full Text
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