同步紅綠藍雙光子螢光顯微術－使用新穎光纖輸出之三色飛秒雷射光源

Abstract

在本論文中，我們提出一種新穎光纖輸出的三色飛秒雷射光源，並利用此光源搭配非線性光學顯微鏡來展示同步紅綠藍雙光子螢光顯微術。此三色光源的產生是藉由一台可攜式1030 nm飛秒雷射作為泵浦源，搭配光子晶體光纖波長轉換器而成。在本實驗中，透過光子晶體光纖能讓原始1030 nm的飛秒脈衝有效地轉換成波段位於600~800 nm的契倫可夫輻射脈衝和波段位於1200~1400 nm的光孤子自頻移脈衝。這三種位於600~800 nm、1030 nm和1200~1400 nm波段的脈衝可以有效地匹配至藍色、綠色和紅色螢光蛋白的雙光子吸收帶。最後，我們將利用此光源來呈現鼠皮毛囊細胞同時染上紅綠藍三色螢光蛋白的雙光子螢光影像。

In this work, we have developed a novel fiber-delivered three-color femtosecond source for nonlinear light microscopy (NLM), and used in simultaneous imaging of three kinds fluorescent proteins (FPs). The high power femtosecond light source was generated by 1030 nm femtosecond pulses as pump and a photonic crystal fiber (PCF) as a compact, flexible, and highly efficient wavelength convertor. By a photonic crystal fiber, the 1030 nm femtosecond pulses were efficiently transformed into energetic cherenkov radiation (CR) pulses in 600~800 nm and soliton self-frequency shift (SSFS) pulses in 1200~1400 nm. These energetic pulses from nonlinear fiber are corresponded to the two-photon absorption band of blue, green and red fluorescent proteins (DAPI, Alexa Fluor 488, and Alexa Fluor 594). Based on the light source, we imply the three-color femtosecond source to excite two photon fluorescence of mouse skin hair follicle cells, expressing a colorful combination of three kinds fluorescent proteins.