Measurement of intrinsic optical backscattering characteristics of cells using fiber-guided near infrared light.
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.
Hsu
Ching-Huang
CH
Institute of Biomedical Engineering, National Cheng Kung University, Tainan, 701 Taiwan, ROC.
Chang
Gwo-Ching
GC
Li
En-Ting
ET
Lin
Yu-Jing
YJ
Chen
Jia-Jin Jason
JJ
D002468:Cell Physiological Phenomena; D048429:Cell Size; D003198:Computer Simulation; D004867:Equipment Design; D019544:Equipment Failure Analysis; D005336:Fiber Optic Technology / Q000295:instrumentation*; D007259:Infrared Rays; D008027:Light; D008954:Models, Biological; D009391:Nephelometry and Turbidimetry / Q000295:instrumentation*; D012031:Refractometry / Q000295:instrumentation*; D012542:Scattering, Radiation