Agilent Cary Eclipse Fluorescence Spectrophotometer

Significance of the topic

Fluorescence and related luminescence techniques are essential tools in analytical chemistry, enabling sensitive detection of a wide range of molecules in life sciences, pharmaceutical research, and quality control. The Agilent Cary Eclipse spectrophotometer, with its xenon flash lamp and versatile accessories, offers a robust platform for rapid, high-sensitivity measurements under ambient conditions, supporting applications from live-cell imaging to thermal stability studies.

Objectives and study overview

This application note illustrates how the Cary Eclipse can be applied across diverse analytical tasks. Key goals include demonstrating the instrument’s fast data collection capabilities, room-light immunity for open-sample manipulations, and cost-effective operation over a long lamp lifetime. A range of real-world examples highlights bio-label characterization, kinetic assays, and stability analyses.

Methodology and instrumentation

• Xenon flash lamp excitation for fluorescence, phosphorescence, chemiluminescence, and bioluminescence measurements without warm-up time.
• Optical design enabling high sensitivity and open sample compartment for reagent additions under room light.
• Rapid scans: full wavelength range in <3 s; single-wavelength kinetics at 80 points/s.
• Accessories: standard quartz cuvette, fast-filter and four-cell Peltier temperature control, powder cell holder, fiber-optic probes, automatic polarizers, microplate reader, and 3D/contour viewing software.

Key results and discussion

• Live-cell imaging and bio-label characterization: fluorescence and absorbance spectra of fluorescein derivatives demonstrated in ELISA and microscopy assays.
• Quantum yield measurement of semiconductor nanocrystals (quantum dots) using standard cuvettes.
• Rapid GPCR oligomerization kinetics: time-based emission analysis shows feasibility of sub-second sampling.
• Bacterial strain detection: probe fluorescence assays under room-light conditions, enabling reagent additions without dark adaptation.
• Intracellular Ca2+ mobilization: 340/380 nm ratio application with 50 ms acquisition intervals for real-time signaling studies.
• Protein tertiary structure and solid-state stability: lyophilized protein analysis with powder cell holder, tracking fluorescence lifetimes of lanthanide chelates.
• Thermal stability of biocatalysts and pharmaceuticals: Tm determination via Peltier-controlled temperature ramps and derivative analysis of fluorescence intensity.

Benefits and practical applications

The Cary Eclipse system delivers:

• High throughput with ultra-fast scans and kinetics for screening and mechanistic studies.
• Cost savings through a xenon lamp life exceeding 10 years.
• Room-light immunity permitting open-chamber additions, ideal for multi-step assays.
• Comprehensive accessory ecosystem for diverse sample formats and temperature-controlled studies.

Future trends and opportunities

Advances in fluorescence instrumentation will likely focus on integration with microfluidic platforms, AI-driven spectral unmixing, expanded detection into near-IR regions, and automated high-content screening. Enhanced software for multi-dimensional data visualization and cloud-based analytics will further extend the Cary Eclipse’s capabilities.

Conclusion

The Agilent Cary Eclipse fluorescence spectrophotometer offers a versatile, user-friendly solution for rapid, sensitive luminescence measurements across life science and industrial applications. Its advanced optical design, long lamp life, and broad accessory compatibility make it a cost-effective workhorse for routine assays and cutting-edge research.

Reference

• Byrne et al. Proc. of SPIE, Vol. 5824, 2005. doi:10.1117/12.604814
• Pellissier et al. J. Biol. Chem., Vol. 286(12), 2011. doi:10.1074/jbc.M110.201939
• Aguirre et al. J. Vis. Exp., Vol. 63, 2012. doi:10.3791/3961
• Petrucci et al. J. Pharmacol. Exp. Ther., Vol. 336, 2011. doi:10.1124/jpet.110.174821
• Ramachandar et al. Anal. Biochem., Vol. 376, 2008. doi:10.1016/j.ab.2008.02.008
• van Lieshout et al. Appl. Biochem. Biotechnol., Vol. 167, 2012. doi:10.1007/s12010-012-9674-z