Abstract：
OBJECTIVE: Genetic reporters encoding fluorescent proteins or luciferase have been used in vivo for the last three decades with claims about their superiority or inferiority over each other. In the present report, a head-to-head in vivo comparison of green fluorescent protein (GFP) fluorescence imaging and luciferase-luciferin imaging, using single-nanometer laser-excitation tuning of fluorescence excitation and an ultra-low-light-detection camera and optics was performed.
METHODS: Mouse Lewis-lung carcinoma cells labeled with GFP (LLC-GFP) or luciferase (LL/2-Luc2) were injected subcutaneously into the flank of nude mice. One week after injection, GFP-fluorescence imaging and luciferase-luciferin imaging was performed using the UVP Biospectrum Advanced system with excitation at 487 nm and peak emission at 513 nm for GFP, and with emission at 560 nm for luciferase-luciferin. GFP fluorescence images were obtained at 0, 10, and 20 min. Luciferase-luciferin images were obtained 10 and 20 min after the injection of D-luciferin.
RESULTS: The intensity of GFP images was 55,909 at 0 min, 56,186 at 10 min, and 57,085 at 20 min, and maintained after 20 min. The intensity of luciferase-luciferin images was 28,065 at 10 min after the injection of D-luciferin and 5,199 at 20 min after the injection. The intensity of luciferase-luciferin images decreased by approximately 80% at 20 min compared to 10 min. An exposure time of 30 s for luciferase-luciferin imaging was needed compared to 100 ms for GFP fluorescence imaging in order to detect signals.
CONCLUSIONS: An imaging system with single-nanometer tuning fluorescence excitation and an ultra-low-light detection camera and optics was able to directly visualize both GFP and luciferase-luciferin images in vivo. The intensity and stability of the signals were both greater for GFP than for luciferase-luciferin, and the exposure time for GFP was 300 times faster, demonstrating the superiority of GFP.
METHODS: Mouse Lewis-lung carcinoma cells labeled with GFP (LLC-GFP) or luciferase (LL/2-Luc2) were injected subcutaneously into the flank of nude mice. One week after injection, GFP-fluorescence imaging and luciferase-luciferin imaging was performed using the UVP Biospectrum Advanced system with excitation at 487 nm and peak emission at 513 nm for GFP, and with emission at 560 nm for luciferase-luciferin. GFP fluorescence images were obtained at 0, 10, and 20 min. Luciferase-luciferin images were obtained 10 and 20 min after the injection of D-luciferin.
RESULTS: The intensity of GFP images was 55,909 at 0 min, 56,186 at 10 min, and 57,085 at 20 min, and maintained after 20 min. The intensity of luciferase-luciferin images was 28,065 at 10 min after the injection of D-luciferin and 5,199 at 20 min after the injection. The intensity of luciferase-luciferin images decreased by approximately 80% at 20 min compared to 10 min. An exposure time of 30 s for luciferase-luciferin imaging was needed compared to 100 ms for GFP fluorescence imaging in order to detect signals.
CONCLUSIONS: An imaging system with single-nanometer tuning fluorescence excitation and an ultra-low-light detection camera and optics was able to directly visualize both GFP and luciferase-luciferin images in vivo. The intensity and stability of the signals were both greater for GFP than for luciferase-luciferin, and the exposure time for GFP was 300 times faster, demonstrating the superiority of GFP.
摘要：
目的：编码荧光蛋白或荧光素酶的基因报告基因在过去的三十年中一直在体内使用，声称它们彼此之间的优势或劣势。在本报告中，绿色荧光蛋白（GFP）荧光成像和荧光素酶-荧光素成像的头对头体内比较，使用单纳米激光激发对荧光激发进行调谐，并进行了超低光检测相机和光学器件。
方法:将用GFP(LLC-GFP)或荧光素酶(LL/2-Luc2)标记的小鼠Lewis-肺癌细胞皮下注射到裸小鼠的侧腹中。注射后一周，GFP-荧光成像和荧光素酶-荧光素成像使用UVPBiospecticAdvanced系统进行，在487nm激发，GFP的峰值发射在513nm，并在560nm处发射荧光素酶-荧光素。在0、10和20分钟获得GFP荧光图像。在注射D-荧光素之后10和20分钟获得荧光素酶-荧光素图像。
结果：0分钟时GFP图像的强度为55,909，在10分钟时56,186，和57,085在20分钟，并在20分钟后保持。荧光素酶-荧光素图像的强度在D-荧光素注射后10分钟为28,065,在注射后20分钟为5,199。与10分钟相比，荧光素酶-荧光素图像的强度在20分钟时降低了约80％。与用于GFP荧光成像的100ms相比,用于荧光素酶-荧光素成像的曝光时间需要30s以检测信号。
结论：具有单纳米调谐荧光激发和超低光检测相机和光学器件的成像系统能够在体内直接可视化GFP和荧光素酶-荧光素酶图像。GFP信号的强度和稳定性均大于荧光素酶-荧光素，GFP的暴露时间快300倍，证明了GFP的优越性。
方法:将用GFP(LLC-GFP)或荧光素酶(LL/2-Luc2)标记的小鼠Lewis-肺癌细胞皮下注射到裸小鼠的侧腹中。注射后一周，GFP-荧光成像和荧光素酶-荧光素成像使用UVPBiospecticAdvanced系统进行，在487nm激发，GFP的峰值发射在513nm，并在560nm处发射荧光素酶-荧光素。在0、10和20分钟获得GFP荧光图像。在注射D-荧光素之后10和20分钟获得荧光素酶-荧光素图像。
结果：0分钟时GFP图像的强度为55,909，在10分钟时56,186，和57,085在20分钟，并在20分钟后保持。荧光素酶-荧光素图像的强度在D-荧光素注射后10分钟为28,065,在注射后20分钟为5,199。与10分钟相比，荧光素酶-荧光素图像的强度在20分钟时降低了约80％。与用于GFP荧光成像的100ms相比,用于荧光素酶-荧光素成像的曝光时间需要30s以检测信号。
结论：具有单纳米调谐荧光激发和超低光检测相机和光学器件的成像系统能够在体内直接可视化GFP和荧光素酶-荧光素酶图像。GFP信号的强度和稳定性均大于荧光素酶-荧光素，GFP的暴露时间快300倍，证明了GFP的优越性。
