Optical, and especially fiber-optic techniques for the sensing of pH have become very attractive and considerable research progress in this field has been made over a number of years. The determination of the pH level across a broad range of applications today, e.g. in life sciences, environmental monitoring, industry and widely in biologically research is now accessible from such optical sensors. This arises because familiar sensors are often limited in terms of their response time and drift, which reduces the use of the current group of such fiber-optic sensors in wider applications. A new compact sensor design has been developed in this work, based on a specially-formed fiber-optic tip that was coated with a pH-sensitive dye, covalently linked to a hydrogel matrix to provide high stability. The sensor developed has a very fast response time (to 90% of saturation, Δt90) of < 5 seconds, a sensing uncertainty of about ± 0.04 pH units and given the covalently bonded nature of the dye, leeching is reduced and the probe is very stable over many days of use. During extended continuous use over ~12h in pH 7, this stability was confirmed, with drift of < 0.05 pH/h. Preliminary experiments in an important biological application, monitoring over pH levels from pH 5 to pH 8.5, are shown and discussed.
While fluorescence-based fiber optic sensors for measuring both pH and oxygen concentration (O2) are well known, current sensors are often limited by their response time and drift, which limits the use of existing fiber optic sensors of this type in wider applications, for example in physiology and other fields. Several new fiber optical sensors have been developed and optimized, with respect to key features such as tip shape and coating layer thickness. In this work, preliminary results on the performance of a suite of pH sensors with fast response times, < 3 second and oxygen sensors (O2) with response times < 0.2 second. The sensors have been calibrated and their performance analyzed using the Henderson–Hasselbalch equation (pH) and classic Lehrer-model (O2).
A focus of research in cell physiology is the detection of Ca2+, NADH, FAD, ATPase activity or membrane potential, only to name a few, in muscle tissues. In this work, we report on a biofluorometer using ultraviolet light emitting diodes (UV-LEDs), optical fibers and two photomultipliers (PMTs) using synchronized fluorescence detection with integrated background correction to detect free calcium, Ca2+, in cardiac muscle tissue placed in a horizontal tissue bath and a microscope setup. Fiber optic probes with imaging optics have been designed to transport excitation light from the biofluorometer's light output to a horizontal tissue bath and to collect emission light from a tissue sample of interest to two PMTs allowing either single excitation / single emission or ratiometric, dual excitation / single emission or single excitation / dual emission fluorescence detection of indicator dyes or natural fluorophores. The efficient transport of light from the excitation LEDs to the tissue sample, bleaching effects of the excitation light in both, polymer and fused silica-based fibers will be discussed. Furthermore, a new approach to maximize light collection of the emission light using high NA fibers and high NA coupling optics will be shown. Finally, first results on Ca2+ measurements in cardiac muscle slices in a traditional microscope setup and a horizontal tissue bath using fiber optic probes will be introduced and discussed.
Optical key parameter of fiber optic flow cells for absorbance measurements are transmission and numerical aperture. In this work we introduce an experimental setup to determine the far field intensity profile of flow cells at different wavelengths in the UV and visible part of the light spectrum. Comparison to spectral transmission will be shown, too.
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