Multiphoton laser surgery at 100 um spatial resolution

100 um 空间分辨率的多光子激光手术

• 批准号: 12450032
• 负责人: NAKAMURA Osamu
• 金额: $ 9.66万
• 依托单位: Osaka University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 2000
• 资助国家: 日本
• 起止时间: 2000 至 2001
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-12450032/
• 关键词: Femtosecond laser; Nonlinear optics; Multiphoton process; Nano biology; Calcium wave; Cell processing; ナノバイオロジー; 多光子吸収; マイクロサージェリー

We totally evaluated the experimental and theoretical knowledge obtained in the first year, from biological (T. Takamatsu) and optical (O. Nakamura) points of view. We optimized the experimental parameters for achieving accurate nanosurgery in biological tissue. We successfully cut and processed red blood cells of rat-cardiomyocyto cells, capillary blood vessels, and others with submicron resolution. Using these results as criterion, we processed the surfaces of rat's and zebrafish's embryo doped with fluorescent protein (GFP) of aequorea coerulescens by genetic manipulation, and evaluated the relation between the processed part and expression of functioning tissue We explored the possibilities of these methods to track the brain growth and to calibrate the structure of embryo to that of an adult brain. Furthermore we investigated total behavior of the manipulated cells using some recent techniques for high resolution including a 4pi confocal optical system. From the investigation, we newly developed a microscope optical system and a laser scanning technique by which we could manipulate cells with 3D resolution and accuracy. Eventually we experimentally studied the relationship between intracellular local breakdown in a nanometer region and the generation of functioning tissue.

我们全面评估了第一年获得的实验和理论知识，从生物（T。Takamatsu）和光学（O.中村）的观点。我们优化了实验参数，以在生物组织中实现精确的纳米手术。我们成功地切割和处理大鼠的红细胞-心肌细胞，毛细血管，和其他亚微米分辨率。以这些结果为标准，我们用基因操作的方法处理了掺杂蓝水母荧光蛋白（GFP）的大鼠和斑马鱼胚胎表面，并评价了处理部位与功能组织表达的关系，探讨了这些方法用于跟踪脑发育和校正胚胎结构与成年脑结构的可能性。此外，我们研究了操纵细胞的总体行为，使用一些最近的高分辨率技术，包括一个4 π共焦光学系统。通过研究，我们新开发了显微镜光学系统和激光扫描技术，通过该技术，我们可以以3D分辨率和精度操纵细胞。最后，我们在实验上研究了纳米区域内细胞内局部击穿与功能组织生成之间的关系。

项目成果

N.Smith, K.Fujita, O.Nakamura and S.Kawata: "Three-dimentional subsurface microprocessing of collagen by ultrashort laser pulses"Applied Physics Letters. 78・7. 999-1001 (2001)

N.Smith、K.Fujita、O.Nakamura 和 S.Kawata：“超短激光脉冲对胶原蛋白进行三维次表面微加工”应用物理快报 78・7（2001 年）。

M.Gu, D.Day, A.O.Nakamura and S.Kawata: "Three-dimensional coherent transfer function for reflection confocal microscopy in the presence of reflective-index mismatch"J. Opt. Soc. Am. A. 18・8. 2002-2006 (2001)

M.Gu、D.Day、A.O.Nakamura 和 S.Kawata：“存在反射折射率失配的三维相干传递函数”J. 18・8。 -2006 (2001)

N.Smith, K.Fujita, O.Nakamura, S.Kawata: "Three-dimentional subsurface microprocessing of collagen by ultrashort laser pulses"Applied Physics Letters. 78・7. 999-1001 (2001)

N.Smith、K.Fujita、O.Nakamura、S.Kawata：“超短激光脉冲对胶原蛋白进行三维次表面微加工”应用物理快报 78・7（2001 年）。

Y.Kawata, M.Murakami, C.Egami, O.Suhihara, N.Okamoto, M.Tsuchimori, O.Watanabe and O.Nakamura: "Nonoptically probing near-field microscopy for the observation of biological living specimens"Applied Physics Letters. 78・15. 2247-2249 (2001)

Y.Kawata、M.Murakami、C.Egami、O.Suhihara、N.Okamoto、M.Tsuchimori、O.Watanabe 和 O.Nakamura：“用于观察生物活体标本的非光学探测近场显微镜”应用物理快报78・15。2247-2249（2001）

K.Fujita,T.Kaneko,O.Nakamura,M.Oyamada,T.Takamatsu and S.Kawata: "Real-time Ca ion wave imaging in living rat-cardiac muscle cells by a confocal multiphoton microscope with a microlens-pinhole array scanner"Proc.of SPIE. 3921・15. 305-312 (2000)

K. Fujita、T. Kaneko、O. Nakamura、M. Oyamada、T. Takamatsu 和 S. Kawata：“通过具有微透镜针孔阵列的共焦多光子显微镜对活体大鼠心肌细胞进行实时 Ca 离子波成像扫描仪”Proc.of SPIE. 3921・15. 305-312 (2000)