**6. References**

470 Recent Advances in Nanofabrication Techniques and Applications

Fig. 15. Two and three-dimensional topography images and cross-section of one of the scratches performed on the HOPG (Sadegh Hassani et al., 2010) , while scanning velocity, number of scratching cycle, time of applying pressure and applied normal force are 10000

This chapter is focused on the study of the Scanning Probe Lithography in a controlled way on various substrates. The load force produced by silicon nitride (NSG11) tip with average spring constant is sufficient to deform and make scratch on the PMMA thin film. The analysis of the roughness of the surface shows that the concept using a thin insulting film of PMMA on silicon and glass surfaces as a scratching mask can be successfully performed for nanopatterning. Drawing patterns are being controlled by the necessary parameters such as normal force, scanning velocity, time of applying pressure and number of scratching cycle. It is shown that the depth of the lithography mark increases linearly with the increase of the applied normal force. The uniformity of scratches on the PMMA coated on silicon and glass

The load force applied by NSG11 tip is not sufficient to exert scratch on the hard surface and is disabled to perform any changes, so diamond tip with much higher spring constant is

nm /s, 1, 1 ms and 50.4 N, respectively.

**5. Conclusion**

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1089.

**24**

*Japan*

**Scanning Probe Lithography**

SunHyung Lee1, Takahiro Ishizaki2, Katsuya Teshima1,

*2National Institute of Advanced Industrial Science and Technology (AIST),*

Lithographic technologies for the surface modification of inorganic and organic surfaces have been developed for various devices such as sensing, data memory, single molecule electronics and biological systems. Nano and micropatterning of organic monolayers have attracted attentions for applications to biological systems in which proteins or DNA are fixed. Photolithography, microcontact printing, and electron beam lithography have usually been used as patterning techniques for organic monolayers (Hayashi et al., 2002; Hong et al., 2003; Saito et al., 2003; Hahn et al., 2004; Kidoaki & Matsuda, 1999). Although the electronbeam lithography can fabricate very small patterns, it requires an ultra-high vacuum system (Harnett et al., 2001). The resolution of photolithography is limited by the light wavelength. Moreover, these methods are based on destructive lithography, i.e., they cause damages to

In particular, nano-lithographic technologies have evolved in order to satisfy persistent demands for miniaturization and high-density integration of semiconductor electric circuits. Scanning probe microscopy (SPM) has been a key tool in achieving this goal. SPM can be used not only as the means that observe surface structure at sub-molecule level by a probe but also as the means that control the atomic and molecular arrangement on a substrate. As a local nano-fabrication means, the lithography technique in nanoscale range by using SPM is called to the scanning probe lithography (SPL) (Kaholek et al., 2004; Blackledge et al., 2000; Tello et al., 2002; Liu et al., 2002). In particular, a variety of lithographic techniques using SPM probe can fabricate nano-scale patterns on an organic monolayer, such as nanoshaving, nanografting, anodization SPL, dip-pen nanolithography (DPN), and electrochemical SPL. The lithography technique is used to break the material surface by using various energy sources. SPM can also be used to break the organic monolayer. For instance, nanoshaving involves mechanical scratching by physical pressure of the probe, and anodization lithography involves anodic oxidation of the substrate surface by an applied bias voltage (above 9 V) between the probe and the substrate (Jang et al., 2002; Kaholek et al., 2004; Sugimura, & Nakagiri, 1995). In the case of the anodization lithography, the oxide layer can be fabricated by anodic oxidation. As another SPL technique, the

**1. Introduction**

the organic materials.

**on Organic Monolayers**

Nagahiro Saito3 and Osamu Takai3 *1Faculty of Engineering, Shinshu University,*

*3EcoTopia Science Institute, Nagoya University,*


*Article* **Establishment of a Newborn Lamb Gut-Loop Model to Evaluate New Methods of Enteric Disease Control and Reduce Experimental Animal Use**

**Ambre Baillou 1,2,†, Nathalie Kasal-Hoc 3,†, Céline Barc 3 , Juliette Cognié 4 , Anne Pinard 3 , Jérémy Pezant 3 , Julie Schulthess 2 , Pauline Peltier-Pain 2 , Sonia Lacroix-Lamandé 1,\* ,‡ and Fabrice Laurent 1,\* ,‡**

**Abstract:** Enteric infectious diseases are not all well controlled, which leads to animal suffering and sometimes death in the most severe cases, in addition to economic losses for farmers. Typical symptoms of enteric infections include watery diarrhea, stomach cramps or pain, dehydration, nausea, vomiting, fever and weight loss. Evaluation of new control methods against enteric infections requires the use of many animals. We aimed to develop a new method for an initial in vivo screen of promising compounds against neonatal diseases such as cryptosporidiosis while limiting experimental animal use. We therefore adapted an in vivo method of multiple consecutive but independent intestinal loops to newborn lambs delivered by cesarean section, in which endotoxin responsiveness is retained. This new method allowed for the screening of natural yeast fractions for their ability to stimulate immune responses and to limit early *Cryptosporidium parvum* development. This model may also be used to investigate host–pathogen interactions and immune responses in a neonatal controlled environment.

**Keywords:** newborn lamb; multiple intestinal loop model; 3R; *Cryptosporidium parvum*; yeast cell wall fractions