**2. Single cell nanosurgery system based on nanorobotic manipulation system**

## **2.1. Single cell nanosurgery system**

We have been proposed a "Nanolaboratory" based on nanorobotic manipulation system from around 2000 [7]. It is one of the systems to realize various nanoscale fabrication and assembly to develop novel nanodevices to integrate borderless technologies based on nanorobotic manipulation system. It is readily applied to the scientific exploration of macroscopic phe‐ nomena and the construction of prototype nanodevices. It would be one of the most significant enabling technologies to realize the manipulation and fabrication technology with individual atoms and molecules for the assembly of devices. Recently, the investigation of Nanoelectro‐ mechanical Systems (NEMS) has attracted much attention [8‐11]. It is expected to realize high integrated, miniaturized, and multi‐functional devices for various applications. One of the effective ways is the direct usage of the bottom‐up fabricated nanostructures.

Nanolaboratory can be applied for the single cell analysis and manipulations. As show in Figure 1, the integration is important for the single cell nanosurgery system between micro and nanorobotic manipulators under various microscopes. The applications under dry, semi wet, and wet conditions can be done under TEM/SEM, E‐SEM and optical microscope (OM). The nanomanipulation system inside TEM/SEM is a fundamental technology for property characterization of nano materials, structures and mechanisms, with the fabrication of nano building blocks, and for the assembly of nano devices. The nanomanipulation system inside E‐SEM provides single cell manipulation and analysis under nano‐scale high resolution images for the application of nanodevices or nanotools assembled under dry condition. OM micromanipulation system is used under water, hence the biological cells can be cultured with medium.

scope. However, its resolution is limited to ~100 nm because of the diffraction limit of optical

**Figure 1.** Single cell nanosurgery system based on micro/nanomanipulators under various microscopes (wet/semi-

Single Cell Nanosurgery System 355

The scanning tunnelling microscopes (STMs) or atomic force microscopes (AFMs), have functions of both observation and manipulation in nano‐scale. Their high resolution makes them capable of atomic manipulation. In 1990, Eigler and Schweize demonstrated that the first atom practice nanomanipulation with scanning tunnelling microscope (STM) [14]. Avouris et al. applied an AFM to bend and translate carbon nanotubes (CNTs) on a substrate [15]. They combined the techniques with an inverse process, namely straightening, by pushing along a bent tube, and realized the translation of a tube to another place. Ning Xi et. al at Michigan State University, developed AFM based nanomanipulation system with interactive operation system [16, 17]. The system realized a real‐time visual feedback during AFM based nanoma‐

Normally, SPM systems have limit for the observation in 2D plane with quite smooth surface. Moreover, the observation area is limited and long time is needed to get one image (more than mins). This limitation rises up as 3D nanomanipulation of nanostructures. On the other hand, the electron microscopes (EMs) provide atomic scale resolution with the electron beam which wave length is less than ~0.1 Å. EMs are divided mainly two types as scanning electron

wavelength (~400 ‐ ~800 nm) explained by the well‐known Abbe's law [13].

nipulation.

wet/dry conditions).

### **2.2. Nanorobotic manipulations**

Nanorobotic manipulation; nanomanipulation, has been received much more attention, because it is an effective strategy for the property characterizations of individual nano‐scale materials and the construction of nano‐scale devices [12]. They might finally be the core‐most part of nanotechnology. One of the attractive future applications of nanomanipulation is to realize the ultimate goal of nanotechnology, or nanomanipulation, is considered.

To manipulate nano‐scale objects, it is needed to observe them with a resolution higher than nano‐scale. Hence, the manipulators and observation systems, microscopes in general, are necessary for nanomanipulations. Figure 1 shows the strategies of nanomanipulations with various kinds of microscopes. The nanomanipulation under various microscopes for 2D/3D nanomanipulations. Optical microscope (OM) is one of the most historical and basic micro‐

bances [6]. Up to now, yeast has maintained its role as a useful model system in fundamental

Micro-Nano Mechatronics — New Trends in Material, Measurement, Control, Manufacturing and Their Applications in

**2. Single cell nanosurgery system based on nanorobotic manipulation**

We have been proposed a "Nanolaboratory" based on nanorobotic manipulation system from around 2000 [7]. It is one of the systems to realize various nanoscale fabrication and assembly to develop novel nanodevices to integrate borderless technologies based on nanorobotic manipulation system. It is readily applied to the scientific exploration of macroscopic phe‐ nomena and the construction of prototype nanodevices. It would be one of the most significant enabling technologies to realize the manipulation and fabrication technology with individual atoms and molecules for the assembly of devices. Recently, the investigation of Nanoelectro‐ mechanical Systems (NEMS) has attracted much attention [8‐11]. It is expected to realize high integrated, miniaturized, and multi‐functional devices for various applications. One of the

Nanolaboratory can be applied for the single cell analysis and manipulations. As show in Figure 1, the integration is important for the single cell nanosurgery system between micro and nanorobotic manipulators under various microscopes. The applications under dry, semi wet, and wet conditions can be done under TEM/SEM, E‐SEM and optical microscope (OM). The nanomanipulation system inside TEM/SEM is a fundamental technology for property characterization of nano materials, structures and mechanisms, with the fabrication of nano building blocks, and for the assembly of nano devices. The nanomanipulation system inside E‐SEM provides single cell manipulation and analysis under nano‐scale high resolution images for the application of nanodevices or nanotools assembled under dry condition. OM micromanipulation system is used under water, hence the biological cells can be cultured with

Nanorobotic manipulation; nanomanipulation, has been received much more attention, because it is an effective strategy for the property characterizations of individual nano‐scale materials and the construction of nano‐scale devices [12]. They might finally be the core‐most part of nanotechnology. One of the attractive future applications of nanomanipulation is to

To manipulate nano‐scale objects, it is needed to observe them with a resolution higher than nano‐scale. Hence, the manipulators and observation systems, microscopes in general, are necessary for nanomanipulations. Figure 1 shows the strategies of nanomanipulations with various kinds of microscopes. The nanomanipulation under various microscopes for 2D/3D nanomanipulations. Optical microscope (OM) is one of the most historical and basic micro‐

realize the ultimate goal of nanotechnology, or nanomanipulation, is considered.

effective ways is the direct usage of the bottom‐up fabricated nanostructures.

studies of disease processes.

Biomedical Engineering

**2.1. Single cell nanosurgery system**

**system**

354

medium.

**2.2. Nanorobotic manipulations**

**Figure 1.** Single cell nanosurgery system based on micro/nanomanipulators under various microscopes (wet/semiwet/dry conditions).

scope. However, its resolution is limited to ~100 nm because of the diffraction limit of optical wavelength (~400 ‐ ~800 nm) explained by the well‐known Abbe's law [13].

The scanning tunnelling microscopes (STMs) or atomic force microscopes (AFMs), have functions of both observation and manipulation in nano‐scale. Their high resolution makes them capable of atomic manipulation. In 1990, Eigler and Schweize demonstrated that the first atom practice nanomanipulation with scanning tunnelling microscope (STM) [14]. Avouris et al. applied an AFM to bend and translate carbon nanotubes (CNTs) on a substrate [15]. They combined the techniques with an inverse process, namely straightening, by pushing along a bent tube, and realized the translation of a tube to another place. Ning Xi et. al at Michigan State University, developed AFM based nanomanipulation system with interactive operation system [16, 17]. The system realized a real‐time visual feedback during AFM based nanoma‐ nipulation.

Normally, SPM systems have limit for the observation in 2D plane with quite smooth surface. Moreover, the observation area is limited and long time is needed to get one image (more than mins). This limitation rises up as 3D nanomanipulation of nanostructures. On the other hand, the electron microscopes (EMs) provide atomic scale resolution with the electron beam which wave length is less than ~0.1 Å. EMs are divided mainly two types as scanning electron

microscopes (SEMs) and transmission electron microscopes (TEMs). For example, M. F. Yu et. al presented the tensile strength of individual CNTs inside a SEM [18]. Howeverthe resolution of SEM, generally ~1 nm resolution, is approximately one order in magnitude lower than that of a TEM. High resolution and transmission image of TEMs are useful for measurement and evaluation of nano‐scale objects. Kizuka et.al proposed the manipulation holder inside high‐ resolution transmission electron microscope (HR‐TEM). The manipulator was specially designed with atomic level positioning resolution [19].

However, the specimen chamber and observation area of TEM are too narrow to contain manipulators with complex functions. Hence, special sample preparation techniques are also needed. We proposed a hybrid nanorobotic manipulation system which is integrated TEM and SEM nanorobotic manipulators as core system for the Nanolaboratory [20, 21]. The strategy is named as hybrid nanomanipulation so as to differentiate it from those with only an exchange‐ able specimen holder. The most important feature of the manipulatoris that it contains several passive DOFs, which makes it possible to perform relatively complex manipulations whereas to keep compact volume to be installed inside the narrow vacuum chamber of a TEM [22].

Recently single cells analysis has been much more attentions because of the progress of the micro/nano scale techniques on the local environmental measurements and controls [23]. Under conventional SEMs and TEMs, the sample chambers of these electron microscopes are set under the high vacuum (HV) to reduce the disturbance of electron beam for observation. To observe water‐containing samples, for example bio‐cells, the appropriate drying and dying treatments are needed before observations. Hence, direct observations of water‐containing samples are normally quite difficult through these electron microscopes.

In this chapter, the novel local stiffness evaluation, local cutting, and local extraction of biological organism are presented by micro‐nanoprobes based on the E‐SEM nanorobotic

Single Cell Nanosurgery System 357

Cell activities, such as embryogenesis, mitosis, morphogenesis, cell orientation, cell motility, and survival depend on attachment to neighboring cells and the extracellular matrix. Cellular attachment to extracellular matrices influences cell morphology, cell function, and signaling mechanisms that direct cellular proliferation and differentiation [29]. Cell‐surface interaction is important in the development of any material or device for biomedical applications, since the performance of a medical device in the body must be compatible with the surrounding tissue [30, 31]. Understanding of the cell adhesion process would benefit the development of

Cell adhesion processes are influenced by numerous parameters, such as the nature of the biomaterial and its surface characteristics (roughness, topography, chemical composition, surface wettability, surface charge and surface treatments) have been investigated. However, the effect of ambient humidity on cell adhesion has had less attention, especially at the single cell level. Understanding the adhesion force at various humidity conditions could help us to better understand the processes of cell‐directed integration during water evaporation. The understanding is also useful in controlling yeast infections at wet environment. Moreover, cell adhesion is influenced by the surface energy of substrate strongly, but the mechanism is still not clear[32]. The study of cell adhesion on substrates with different surface energy could help

manipulation system for future cell diagnosis and surgery system.

**Figure 2.** Single Cell Nano-surgery System with various nano-tools

**3.1. Adhesion force measurement of single cell using nano‐putter**

suitable biomaterials or device for both tissue engineering and medical fields.

us to understand the adhesion mechanism better.

On the other hand, the environmental‐SEM (E‐SEM) can be realized the direct observation of water‐containing samples with nanometer high resolution by specially built secondly electron detector [24]. The evaporation of water is controlled by the sample temperature (~0 ‐ ~40 °C) and sample chamber pressure (10 ‐ 2600 Pa). The unique characteristic of the E‐SEM is the direct observation of the hydroscopic samples with non‐drying treatment. Hence, the nano‐ manipulation inside the E‐SEM is considered to be an effective tool for a water‐containing sample with nanometer resolution [25‐28].
