Multi-wavelength femtosecond laser sources for intravital multiphoton microscopy

用于活体多光子显微镜的多波长飞秒激光源

• 批准号: 8852123
• 负责人: Charles P. Lin
• 金额: $ 60.2万
• 依托单位: CORNELL UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-08-01 至 2016-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8852123
• 关键词: Address; Amplifiers; Animals; Area; Back; Biological; Bone Marrow; Cells; Clinical; Color; Engraftment; Fiber; Frequencies; Generations; Goals; Health; Hematopoietic Neoplasms; Hematopoietic Stem Cell Transplantation; Hematopoietic stem cells; Homing; Image; Imaging technology; Individual; Industry; Label; Lasers; Lead; Life; Light; Lymphoma; Methods; Microscope; Microscopy; Monitor; Mus; Optics; Osteoblasts; Patients; Physiologic pulse; Procedures; Pulse Rates; Recovery; Research; Research Personnel; Semaphorins; Source; Speed; Stem Cell Research; Stem cell transplant; Stem cells; System; Techniques; Technology; Telecommunications; Testing; Tissues; Transplantation; base; bioimaging; cell type; cohort; cost; design; fluorophore; imaging modality; improved; in vivo; in vivo imaging; innovation; insight; intravital imaging; leukemia; lipid biosynthesis; meetings; novel; programs; regenerative; research study; sapphire laser; second harmonic; telecom-wavelength

DESCRIPTION (provided by applicant): This is a collaborative research program that brings together an optical technology group (Dr. Xu) and an in vivo imaging group (Dr. Lin). The two groups share a common goal to develop imaging technology for solving biomedical problems and addressing clinical needs. Here we focus on the need to improve hematopoietic stem cell (HSC) homing and engraftment after HSC transplantation (HSCT). This life-saving procedure is often the last hope of cure for patients with cancers of the blood system such as leukemia or lymphoma, but successful transplantation can be achieved only if a sufficient number of transplanted HSCs are able to reach and engraft the patient's bone marrow (BM). To help improve stem cell homing and engraftment, the Lin group has developed intravital imaging methods to track individual HSCs in the BM of live animals after transplantation. However, the current view of the BM microenvironment is severely limited due to the inadequacies of the available imaging technology. To gain a more comprehensive view of the BM microenvironment, where multiple cell types interact and form a supportive niche for HSC engraftment, the Xu group will develop a novel fiber-based source for nonlinear microscopy, which will enable simultaneous imaging of multiple fluorescent indicators as well as enabling label-free harmonic generation and vibrational imaging. Integration of the new source with the intravital microscope will enable the Lin group to proceed with experiments that had been envisioned but were held back due to lack of a suitable technology. The proposed source is based on the following innovations: (1) Soliton self-frequency shift (SSFS) in a large mode area (LMA) fiber enables the generation of energetic, widely wavelength tunable soliton pulses seeded from a fiber laser at the telecom wavelength, and the subsequent second harmonic generation (SHG) of the fundamental wavelength enables a single turn-key, low-cost, fiber-based source to generate three independent wavelength tunable sources to excite multiple fluorophores. 2) All-fiber, high-speed intensity modulation to electronically control the wavelength, repetition rate, and pulse delay. 3) A single light source will enable experiments that currently require two synchronized Ti:sapphire lasers plus an optical parametric oscillator (OPO) and a regenerative amplifier. Leveraging the highly mature and integrated techniques that have been developed for the telecommunications industry, we aim to create a "telecom grade" femtosecond source that is truly robust and versatile. The versatility is important for tailoring the source to meet specific imaging needs while the robustness is essential for the biological studies that require longitudinal imaging of large cohorts of animals. The successful completion of this program will not only advance imaging technology but also advance stem cell research. In addition, the technology will be broadly applicable and will significantly increase the accessibility of femtosecond sources to other biomedical researchers.

描述（由申请人提供）：这是一个协作研究计划，将光学技术组（XU博士）和一个体内成像组（Lin博士）汇集在一起​​。这两组共享一个共同的目标，以开发成像技术来解决生物医学问题并满足临床需求。在这里，我们关注的是需要改善HSC移植后（HSCT）后造血干细胞（HSC）的归巢和植入。对于血液系统（例如白血病或淋巴瘤）患者来说，这种挽救生命的过程通常是治愈的最后希望，但是只有在足够数量的移植HSC能够接触并植入患者的骨髓（BM）时，才能成功地进行移植。为了帮助改善干细胞寄养和植入，LIN组开发了浸泡成像方法，以跟踪移植后活体动物BM中的单个HSC。但是，由于可用成像技术的不足，BM微环境的当前观点受到严重限制。为了获得BM微环境的更全面的视图，其中多种细胞类型相互作用并形成了HSC植入的支持性利基市场，XU组将开发出基于非线性显微镜的新型纤维来源，这将启用多个荧光指示剂的同时成像，并启用标记无标记的和无关的和振动的影像。新来源与插入显微镜的集成将使Lin组能够进行已设想的实验，但由于缺乏合适的技术而被阻止。 The proposed source is based on the following innovations: (1) Soliton self-frequency shift (SSFS) in a large mode area (LMA) fiber enables the generation of energetic, widely wavelength tunable soliton pulses seeded from a fiber laser at the telecom wavelength, and the subsequent second harmonic generation (SHG) of the fundamental wavelength enables a single turn-key, low-cost, fiber-based产生三个独立波长可调源以激发多个荧光团的来源。 2）全纤维，高速度调制，以电子控制波长，重复率和脉冲延迟。 3）单个光源将启用当前需要两个同步的Ti：蓝宝石激光器以及光学参数振荡器（OPO）和再生放大器的实验。利用为电信行业开发的高度成熟和集成的技术，我们旨在创建一个真正强大且多功能的“电信等级” femtsecond。多功能性对于调整来源以满足特定的成像需求很重要，而鲁棒性对于需要大量动物的纵向成像的生物学研究至关重要。该程序的成功完成不仅将推进成像技术，还将推进干细胞研究。此外，该技术将广泛适用，并将大大提高飞秒来源对其他生物医学研究人员的可访问性。