本論文一開始，以孟加拉玫瑰素(rose Bengal，RB)當作光起始劑之多光子激發(multiphoton excitation，MPE)微製作技術被用來製作各種材料之二維微米圖案及三維微米結構，例如細胞外基質(extracellular matrix，ECM)蛋白質、明膠和高分子聚合物，並利用它們來引導小鼠大腦皮質神經細胞的軸突生長。實驗結果顯示，神經軸突的生長可以很有效率的被侷限或引導藉由這些大面積的二維正向和負向微米圖案，同時也探討了不同材料對於神經軸突生長的影響。除此之外，我們亦使用了另一種變性膠原蛋白材料－明膠，或明膠與多聚賴氨酸(poly-D-lysine，PDL)的混合液來製作具生物相容性的三維微米結構。其結果亦顯示這些微結構對於引導神經軸突在三維環境的生長是很有潛力的。
然而，由於傳統的MPE微製作技術是採用點掃描的方式來進行，因此、製作速度慢所造成的低產量是其最大缺點。為了改善這個限制，我們發展了一套具時域聚焦與圖形化照射的MPE微製作系統。藉由一層一層堆疊的方式，任意形狀的三維高分子聚合物結構可以被快速地製作出來。與傳統的掃描式MPE相比，此方法不僅可製作出任意形狀的微結構，在製作的速度上更可提升約三個數量級。此外，這套系統整合了一個數位微面鏡裝置(digital micromirror device)可用來針對特定的區域，局部地控制雷射脈衝的數量。藉由這項功能並搭配RB當作光活化劑，多個不同灰階(梯度濃度)的三維牛血清蛋白(bovine serum albumin，BSA)結構可以在幾秒鐘之內被同時製作出來。同時，所製作出來的牛血清蛋白結構亦可被即時監測藉由利用RB當作雙光子激發螢光(two-photon excited fluorescence，TPEF)的對比劑。因具有獨特的高速、灰階製作以及即時監控的能力，此方法符合了許多生醫應用的需求，特別是對於細胞外基質的相關研究。
最後，我們亦利用此系統對氧化石墨烯(graphene oxide，GO)薄膜進行高通量多光子誘發還原與剝離的研究。還原的程度以及剝離的過程可以很精準地被操控藉由調控雷射的波長、功率以及脈衝數量。相對於逐點雷射掃描的方式，此方法具備了高產能與多功能的優點，進而可以在GO的薄膜上製作出具有微米特徵尺度的大面積圖案。因此，此具有時域聚焦與脈衝控制的超快雷射系統可符合高產量的需求，將有助於GO相關材料在微電子裝置上的應用。

In this thesis, multiphoton excitation (MPE) microfabrication using Rose Bengal (RB) as the photoinitiator was first used to create two-dimensional (2D) micropatterns and three-dimensional (3D) microstructures with various materials, such as extracellular matrix (ECM) proteins, gelatin, and polymer, and then utilized the bio-microstructures to guide axonal outgrowth of primary cortical neurons. Large-scale positive and negative patterns were created, and the results demonstrated that neurite growth can be constrained or guided in a well-controlled manner via the micropatterns. In addition, gelatin, a denature collagen or mixture of gelatin and poly-D-lysine (PDL) were used for fabricating 3D microstructures. The results are also promising for guiding growth of axons in 3D microenvironment.
However, one of the limits of conventional scanning MPE microfabrication is its low throughput due to point-by-point processing. To surpass this limit, a MPE microfabrication system based on temporal focusing and patterned excitation has been developed to quickly provide 3D freeform polymer microstructures. The microstructures are created by sequentially stacking 2D fabricating patterns. Compared to conventional scanning MPE, this approach offers freeform microstructures and a greater than three-order increase in fabrication speed. Furthermore, the system integrated with a digital micromirror device for locally controlling the laser pulse numbers which can provide high-throughput fabrication of 3D gray-level bio-microstructures via two-photon crosslinking. Multiple bovine serum albumin (BSA) structures of different concentrations were simultaneously achieved by selecting different pulse numbers in the designated regions with an appropriate femtosecond laser power within a few seconds. Moreover, the fabricated BSA microstructure can be monitored online by utilizing the RB two-photon excited fluorescence (TPEF) as contrast agent. This approach with its unique capability of high-speed, gray-level, and online-inspection fabrication meets the requirements of the biomedical researches involved in ECM.
Finally, high-throughput multiphoton-induced reduction and ablation of graphene oxide (GO) films has also been studied using this system. The degree of reduction and ablation can be precisely implemented via controlling the laser wavelength, power, and pulse number. Compared to point-by-point scanning laser direct writing, this method offers a high-throughput and multiple-function approach to accomplish large-area and micro-scale patterns on GO films. The high-throughput micropatterning of GO via the temporal focusing-based femtosecond laser system matches the requirement of mass production for GO-based applications in microelectronic devices.

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