Conventional flow cytometry generally uses single-photon excitation of fluorescence from cells or other particles. By labeling cells with multiple dyes, and by using size measurements via the elastically scattered excitation light, multiple-parameter measurements can be made. The tremendous power of this technique has made it the standard technique to quantify fluorescence in cells. For multiple-dye, multi-wavelength analysis, multiple excitation sources (i.e. lasers) may be required. In light of this arrangement, the layout of such devices can be complex, adding to the already-existing complexity of its fluid flow system, which put limits on their applications.
Researchers at the University of Michigan have developed a multiphoton flow cytometer is provided that has a single laser source for excitation of multiple dyes and does not require sheath flow conditions. Thus, the multiphoton flow cytometer not only has a simple configuration, but also opens up a wide range of new applications, such as virus detection and in vivo measurements. In particular, it takes advantage of double-clad photonic crystal fiber optic technology, which provides a significant performance improvement. This photonic crystal fiber has a unique microstructure, which ensures an efficient delivery of excitation laser beam for a high excitation rate and high resolution, while enables one to achieve real optimization of signal collection efficiency at the same time. Using this fiber, detection sensitivity of more than 200 fold can be achieved, compared to the commercially available single-mode fiber.
Applications and Advantages
- Quantitative Two-Photon Flow Cytometry
- Enhanced sensitivity
- Simple configuration
- Amenable to wide range of new applications