Mathew Alexander S, DeRosa Christopher A, Demas James N, Fraser Cassandra L
Department of Chemistry, University of Virginia, Charlottesville, Virginia 22904, USA.
Anal Methods. 2016 Apr 21;8(15):3109-3114. doi: 10.1039/C5AY02959G. Epub 2016 Apr 1.
Lifetime-based oxygen imaging is useful in many biological applications but instrumentation can be stationary, expensive, and complex. Herein, we present a portable, cost effective, simple alternative with high spatiotemporal resolution that uses a complementary metal oxide silicon (CMOS) camera to measure oxygen sensitive lifetimes on the millisecond scale. We demonstrate its compatibility with difluoroboron β-diketonate poly(lactic acid) (BFbdkPLA) polymers which are nontoxic and exhibit long-lived oxygen sensitive phosphorescence. Spatially resolved lifetimes of four BFbdkPLA variants are measured using nonlinear least squares (NLS) and rapid lifetime determination (RLD) both of which are shown to be accurate and precise. Real-time imaging in a dynamic environment is demonstrated by determining lifetime pixel-wise. The setup costs less than $5000, easily fits into a backpack, and can operate on battery power alone. This versatility combined with the inherent utility of lifetime measurements make this system a useful tool for a wide variety of oxygen sensing applications. This study serves as an important foundation for the development of dual mode real time lifetime plus ratiometric imaging with bright, long lifetime difluoroboron β-diketonate probes.
基于寿命的氧成像在许多生物学应用中很有用,但仪器设备可能是固定的、昂贵的且复杂的。在此,我们展示了一种具有高时空分辨率的便携式、经济高效且简单的替代方案,该方案使用互补金属氧化物硅(CMOS)相机在毫秒尺度上测量对氧敏感的寿命。我们证明了它与二氟硼β - 二酮聚乳酸(BFbdkPLA)聚合物的兼容性,该聚合物无毒且表现出长寿命的氧敏感磷光。使用非线性最小二乘法(NLS)和快速寿命测定法(RLD)测量了四种BFbdkPLA变体的空间分辨寿命,结果表明这两种方法都准确且精确。通过逐像素确定寿命来展示在动态环境中的实时成像。该装置成本低于5000美元,易于装进背包,并且仅靠电池供电就能运行。这种多功能性与寿命测量的内在实用性相结合,使该系统成为各种氧传感应用的有用工具。这项研究为开发具有明亮、长寿命二氟硼β - 二酮探针的双模式实时寿命加比率成像奠定了重要基础。