本論文主要是利用飛秒(1 fs =10-15 s)雷射對果蠅頭進行剝離加工工作，此類非均質的生物組織的外殼組成為甲殼素，學名為幾丁質，其殼是由表皮分泌而成，與內部軟組織相互連結，從頭殼表面開始，由外往內來看，明顯呈現非均質的組織架構，為了可以更進一步研究內層的腦組織，因此需去除表面頭殼，故在加工上面的研究是找出幾丁質的最適當加工條件，使內層組織可為生物研究使用。
飛秒雷射透過色散補償產生極短的脈衝寬度，在雷射光的聚焦處，可以產生強大的峰值功率，本論文中使用的次飛秒雷射系統，在脈衝寬度＜50 fs的條件下，透過高數值孔徑物鏡的聚焦，在聚焦處產生非線性效應與雙光子激發，配合三維平台的掃描系統與透過LabVIEW程式控制的飛秒雷射加工影像系統來調整參數，作為加工的工具，期望達到更細緻的加工結果，包含切割邊緣的銳利度與降低熱效應對非加工區域組織的破壞與汙染，以螢光影像作為驗證加工效果是否符合預期的依據，以實驗結果來看，確實得到了期望的成果，成功為非均質生物組織的研究新增另一個加工工具的選項，並帶來更便利與副作用更少的優點。

In this thesis, a developed femtosecond (1 fs =10-15 s) laser system is utilized to process drosophila’s head. The inhomogeneous biological tissue is covered by a shell which is constructed by chitonsan, scientific name is chitin. The shell is secreted by the epidermis and interconnected with the internal soft tissue. To consider the cross-section of drosophila’s head, the organization structure is heterogeneity definitely. In order to further study the inner brain tissue, the shell must be partially removed first. Therefore, the study of the drosophila machining is to investigate and optimal machining processing conditions for chitin to keep the inner tissue good enough for biological study.
The advantage of femtosecond laser is that can processing with very low average power and very short pulse duration closing to the femtosecond or even sub-femtosecond in the focal point and result in strong peak power. In this thesis, the femtosecond laser system with the pulse width under 50 fs and through a high numerical aperture objective interacted with materials in local focal point may induce the nonlinear effect and two-photon excitation. In conjunction with three-dimensional scanning stage controlled by LabVIEW program and adjust the parameters of femtosecond laser machining processing and imaging system to achieve a more detailed machining result eventually. Through two photon excitation images, the improvements of machining results including edge sharpness, low thermal effect, and less contaminant are demonstrated.

1.張家禎、張慧羽，“果蠅在遺傳學上的貢獻”，科學發展月刊481，6-11 (2013)。
2.FlyBase, http://flybase.org/.
3.FlyMove, http://flymove.uni-muenster.de/.
4.M. Goppert-Mayer, “Elementary file with two quantum fissures,” Ann. Phys. 9, 273–294 (1931).
5.I. Freund and M. Deutsch, “2nd-harmonic microscopy of biological tissue,” Opt. Lett. 11, 94-96 (1986).
6.W. Denk, J. H. Strickler, and W. W. Webb, “Two-photon laser scanning fluorescence microscopy,” Science 248, 73–76 (1990).
7.李宜真、蘇子正，“雙光子顯微術於生物組織的應用”，物理雙月刊24，413 (2001)。
8.M. H. Niemz, Laser-Tissue Interactions-Fundamentals and Applications, Springer-Verlag, 45-149 (2007).
9.J. L. Boulnois, “Photophysical processes in recent medical laser developments: a review,” Lasers Med. Sci. 1, 47–66 (1986).
10.P. Teng, N. S. Nishioka, R. R. Anderson, and T. F. Deutsch, “Acoustic studies of the role of immersion in plasma-mediated laser ablation,” IEEE J. Qu. Electron. QE-23, 1845–1852 (1987).
11.D. Stern, R. W. Schoenlein, C.A. Puliafito, E. T. Dobi, R. Birngruber, and J. G. Fujimoto, “ Corneal ablation by nanosecond, picosecond and femtosecond lasers at 532nm and 625 nm,” Arch. Ophthalmol. 107, 587–592 (1989).
12.M. H. Niemz, E. G. Klancnik, and J. F. Bille, “Plasma-mediated ablation of corneal tissue at 1053nm using a Nd:YLF oscillator/regenerative amplifier laser,” Lasers Surg. Med. 11, 426–431 (1991).
13.C. A. Puliafito and R. F. Steinert, “Short-pulsed Nd:YAG laser microsurgery of the eye: biophysical considerations,” IEEE J. Qu. Electron. QE-20, 1442–1448 (1984).
14.A. Vogel, J. Noack, G. Huttman, and G. Paltauf, “Mechanism of femtosecond laser nanosurgery of cells and tissues”, Appl. Phys. 81, 1015–1047 (2005).
15.李益誠，利用無孔式進場掃描式光學顯微術於電漿子結構之研究，國立成功大學，九十八年度碩士論文。
16.KMLabs, http://www.kmlabs.com/.
17.H. M. Carol, T. K. William, Biology-An Exploration of Life, W. W. Norton & Company, 522-539 (1996).
18.BioBookDiversity,http://mac122.icu.ac.jp/biobk/BioBookDiversity_8.html.
19.Jewel beetle, http://www.everythingabout.net.
20.王嘉薇，丁醯化幾丁聚醣之研究，國立成功大學，九十二年度碩士論文。
21.F. R. Turner and A. P. Mahowald “Scanning electron microscopy of Drosophila melanogaster embryogenesis. III. Formation of the head and caudal segments,” Dev. Biol. 68, 96–109 (1979).
22.Newport, Measuring pulsed beams with a slit-based profilr, http://www.photon-inc.com.