Agilent Cary Eclipse Fluorescence Spectrophotometer

Importance of the Topic

The Agilent Cary Eclipse Fluorescence Spectrophotometer provides a versatile platform for measuring fluorescence, phosphorescence, chemiluminescence and bioluminescence with minimal photobleaching and high immunity to ambient light. Its rapid data acquisition and long‐lifetime xenon flash lamp make it an essential tool in life sciences research, pharmaceutical stability studies, and industrial quality control.

Objectives and Overview

This summary highlights the key applications and performance features of the Cary Eclipse instrument as reported in various peer‐reviewed studies. The main goals include:

• Characterizing fluorescent bio‐labels and nanocrystals
• Monitoring GPCR oligomerization kinetics and bacterial detection assays
• Measuring intracellular ion mobilization
• Analyzing protein structural changes and thermal stability
• Demonstrating the flexibility of accessory modules

Instrumentation Used

The core system and accessories described in the literature include:

• Cary Eclipse Fluorescence Spectrophotometer with xenon flash lamp
• Standard quartz cuvettes and powder cell holders for solid‐state samples
• Fast filter accessory for sub‐50 ms kinetics
• Four‐cell Peltier temperature controller for thermal ramp experiments
• Fiber‐optic probes and microplate reader adapter
• Automatic polarizer and fast emission filters
• Cary WinFLR software with dedicated ratio and 3D/contour viewing modules

Methodology

Fluorescence and luminescence measurements were performed with the sample compartment open to avoid photo‐bleaching. Full‐range wavelength scans (<3 s) and single‐wavelength kinetics (80 points/s) were used for rapid assays. Intracellular calcium dynamics were monitored using the 340/380 nm excitation ratio method. Solid‐state protein samples were studied in a powder cell holder to assess long‐term storage stability. Temperature‐dependent fluorescence changes and melting transitions (Tm) were obtained via controlled thermal ramping of the Peltier accessory.

Main Results and Discussion

Key findings from representative experiments include:

• Fluorescein derivatives showed clear absorbance/excitation and emission spectra suitable for ELISA and microscopy assays.
• Room light immunity allowed addition of reagents during bacterial detection assays without background interference.
• GPCR oligomerization kinetics were resolved in real time using emission spectral analysis.
• Intracellular Ca²⁺ mobilization was quantified by the 340/380 nm ratio in live cells at 50 ms intervals.
• Lyophilized protein stability and fluorescence lifetimes of lanthanide chelates were characterized under solid‐state conditions.
• Thermal denaturation of a fluorescently labeled PNA probe yielded a Tm of 63.9 °C via derivative analysis.

Benefits and Practical Applications

The Cary Eclipse system offers:

• High temporal resolution for kinetic studies
• Low photobleaching risk with open‐sample design
• Long lamp lifetime (>10 years) and minimal maintenance costs
• Modular accessory options for diverse assays
• Software tools for ratio analysis, 3D mapping, and thermal transitions

Future Trends and Opportunities

Emerging directions for fluorescence instrumentation include:

• Integration with machine‐learning algorithms for automated spectral analysis
• Miniaturized and portable designs for field and point‐of‐care testing
• Enhanced time resolution for ultrafast photophysical studies
• Multiplexed assays and microfluidic platforms to increase throughput
• In vivo fluorescence imaging and fiber‐optic deployment in live tissues

Conclusion

The Agilent Cary Eclipse Fluorescence Spectrophotometer combines robust optical design, rapid data acquisition, and a broad range of accessories to meet the demands of modern analytical chemistry. Its performance across life science assays, protein stability studies, and industrial QA/QC underscores its role as a versatile workhorse in fluorescence measurement.

References

• Byrne, J. D., et al. (2005) Measurement of quantum yield in nanocrystals using a standard quartz cuvette. Proc. SPIE 5824. doi:10.1117/12.604814
• Pellissier, L. P., et al. (2011) Rapid kinetics of GPCR oligomerization in live cells. J. Biol. Chem. 286(12). doi:10.1074/jbc.M110.201939
• Aguirre, S., et al. (2012) Fluorescent assay for bacterial strain detection. J. Vis. Exp. 63. doi:10.3791/3961
• Petrucci, R., et al. (2011) Intracellular calcium mobilization using ratio analysis. J. Pharmacol. Exp. Ther. 336. doi:10.1124/jpet.110.174821
• Ramachandar, R., et al. (2008) Solid‐state fluorescence analysis of lyophilized proteins. Anal. Biochem. 376. doi:10.1016/j.ab.2008.02.008
• van Lieshout, M., et al. (2012) Thermal stability of biocatalysts via fluorescence ramp. Appl. Biochem. Biotechnol. 167. doi:10.1007/s12010-012-9674-z