Measurement of intrinsic optical backscattering characteristics of cells using fiber-guided near infrared light

Abstract

Background: Intrinsic optical signals (IOS), which reflect changes in transmittance and scattering light, have been applied to characterize the physiological conditions of target biological tissues. Backscattering approaches allow mounting of the source and detector on the same side of a sample which creates a more compact physical layout of device. This study presents a compact backscattering design using fiber-optic guided near-infrared (NIR) light to measure the amplitude and phase changes of IOS under different osmotic challenges.Methods: High-frequency intensity-modulated light was guided via optic fiber, which was controlled by micromanipulator to closely aim at a minimum cluster of cortical neurons. Several factors including the probe design, wavelength selection, optimal measuring distance between the fiber-optical probe and cells were considered. Our experimental setup was tested in cultured cells to observe the relationship between the changes in backscattered NIR light and cellular IOS, which is believed mainly caused by cell volume changes in hypo/hyperosmotic solutions (± 20, ± 40 and ± 60 mOsm).Results: The critical parameters of the current setup including the optimal measuring distance from fiber-optical probe to target tissue and the linear relationship between backscattering intensity and cell volume were determined. The backscattering intensity was found to be inversely proportional to osmotic changes. However, the phase shift exhibited a nonlinear feature and reached a plateau at hyperosmotic solution.Conclusions: Our study indicated that the backscattering NIR light guided by fiber-optical probe makes it a potential alternative for continuous observation of intrinsic optical properties of cell culture under varied physical or chemical challenges. © 2010 Hsu et al; licensee BioMed Central Ltd.