Photonics Group, Department of Physics, Imperial College London, London, SW7 2AZ, UK.
Kentech Instruments Ltd., Howbery Park, Wallingford, OX10 8BD, UK.
J Fluoresc. 2017 Sep;27(5):1643-1654. doi: 10.1007/s10895-017-2101-7. Epub 2017 May 25.
Autofluorescence lifetime measurements, which can provide label-free readouts in biological tissues, contrasting e.g. different types and states of tissue matrix components and different cellular metabolites, may have significant clinical potential for diagnosis and to provide surgical guidance. However, the cost of the instrumentation typically used currently presents a barrier to wider implementation. We describe a low-cost single point time-resolved autofluorescence instrument, exploiting modulated laser diodes for excitation and FPGA-based circuitry for detection, together with a custom constant fraction discriminator. Its temporal accuracy is compared against a "gold-standard" instrument incorporating commercial TCSPC circuitry by resolving the fluorescence decays of reference fluorophores presenting single and double exponential decay profiles. To illustrate the potential to read out intrinsic contrast in tissue, we present preliminary measurements of autofluorescence lifetime measurements of biological tissues ex vivo. We believe that the lower cost of this instrument could enhance the potential of autofluorescence lifetime metrology for clinical deployment and commercial development.
自动荧光寿命测量,可在生物组织中提供无标记的读数,例如对比不同类型和状态的组织基质成分和不同的细胞代谢物,可能对诊断和提供手术指导具有重要的临床潜力。然而,目前常用的仪器成本通常是广泛应用的障碍。我们描述了一种低成本的单点时间分辨自动荧光仪器,利用调制激光二极管进行激发和基于 FPGA 的电路进行检测,以及定制的恒比甄别器。通过解析呈现单指数和双指数衰减曲线的参考荧光团的荧光衰减,将其时间精度与包含商业 TCSPC 电路的“黄金标准”仪器进行比较。为了说明在组织中读取固有对比度的潜力,我们展示了生物组织离体自动荧光寿命测量的初步测量结果。我们相信,该仪器的成本降低将增强自动荧光寿命计量在临床部署和商业开发方面的潜力。