Optical sensor microscopy

The resolution of a conventional microcope is limited by the classical phenomena of interference and diffraction. The limit is approximately X/2, X being the wavelength. This limit can be overcome by using a sub-wavelength light source and by placing the sample very close to this source (i.e. in the near field). The relevant domain is near-field optics (as opposed to far-field conventional optics), which has been applied to microscopy, spectroscopy and optical sensors. In particular, nearfield scanning optical microscopy (N SOM) has proved to be a powerful tool in physical, chemical and life sciences (Dunn, 1999). 

Perhaps the most well-recognized fluorescent dye for detection of DNA hybridization is ethidium bromide (EtBr). EtBr is a cationic phenanthridinium compound that can bind to DNA by intercalation. This dye has an excitation maxima at 518 nm when bound to double-stranded DNA (dsDNA). Excitation of EtBr is often done by use of an argon ion laser, making this fluorophore a viable choice for applications in optical sensors as well as confocal scanning laser microscopy and fluorometry [41]. The structure of ethidium bromide is shown in Fig. 6. 

Dodecyl acrylate microspheres, crosslinked with hexanediol diacrylate, were prepared by emulsion polymerisation and characterised by scanning electron microscopy, particle size measurement, and differential scanning calorimetry. The microspheres were doped with 9-(diethylamino)-5-(octadecanoylimino)-5H-benzo(alpha)phenoxazine (ETH 5294), a chromoionophore whose fluorescence emission spectrum is sensitive to pH. These microspheres adsorb anions selectively according to their lipophilicity, and therefore can be used as anion-selective optical sensors. The responses of the microspheres to chloride, nitrate and perchlorate ions were reported. 27 refs. 

Fixed focal length microlenses and arrays with lens diameters of a few to several hundred micrometers are extensively used in many optical systems such as projection lithography [1-3], optical coherence microscopy (OCM) [4], Shack-Hartmann sensors [5], and back lighting for projection liquid crystal displays (LCDs) [6]. 

A most recent commercial Nano Indenter (Nano Indenter XP (MTS, 2001)) consists of three major components [66] the indenter head, an optical/atomic force microscope, and x-y-z motorized precision table for positioning and transporting the sample between the optical microscopy and indenter (Fig. 28). The load on the indenter is generated using a voice coil in permanent magnet assembly, attached to the top of the indenter column. The displacement of the indenter is measured using a three plate capacitive displacement sensor. At the bottom of the indenter rod, a three-sided... 

In addition to absolute pressure measurements, pressure sensors can be used to determine flow rates when combined with a well-defined pressure drop over a microfluidic channel. Integration of optical waveguide structures provides opportunities for monitoring of segmented gas-liquid or liquid-liquid flows in multichannel microreactors for multiphase reactions, including channels inside the device not accessible by conventional microscopy imaging (Fig. 2c) (de Mas et al. 2005). Temperature sensors are readily incorporated in the form of thin film resistors or simply by attaching thin thermocouples (Losey et al. 2001). 

Fluorescent pH indicators offer much better sensitivity than the classical dyes such as phenolphthalein, thymol blue, etc., based on color change. They are thus widely used in analytical chemistry, bioanalytical chemistry, cellular biology (for measuring intracellular pH), medicine (for monitoring pH and pCC>2 in blood pCC>2 is determined via the bicarbonate couple). Fluorescence microscopy can provide spatial information on pH. Moreover, remote sensing of pH is possible by means of fiber optic chemical sensors. 

Several papers have been published in which, instead of concentrating on specific reactions, the technology was highlighted. One, by Marose et al.,7 discusses the various optics, fiber optics, and the probe designs that allow in situ monitoring. They describe the various optical density probes used for biomass determination in situ microscopy, optical biosensors, and specific sensors such as NIR and fluorescence. 

Applications of fiber-optic pH sensors in environmental analysis, biomedical research, medical monitoring, and industrial process control have been reviewed by Lin [67]. A multitude of luminescent systems for pH monitoring are commercially available, mostly under special trademarks. Pyrene [68-70], coumarin, bromothymol blue [71] and fluorescein [72-74] derivatives are typical examples that have been used in research in the past two decades. Carboxyfluorescein derivatives have been directly applied to skin tissue samples for the lifetime imaging of pH gradients in the extracellular matrix of the epidermis [75]. Two-photon excitation microscopy became an estab-... 

Fluorescence sensors for saccharides are of particular interest in a practical sense. This is in part due to the inherent sensitivity of the fluorescence technique. Only small amounts of a sensor are required (typically 10-6 M), offsetting the synthetic costs of such sensors. Also, fluorescence spectrometers are widely available and inexpensive. Fluorescence sensors have also found applications in continuous monitoring using an optical fiber and intracellular mapping using confocal microscopy. 

Therefore, local dissolution and recrystallization seem to play an important role in the gas uptake mechanism in these type of sensor materials. The coordination of SO2 to the platinum center (and the reverse reaction) is therefore likely to take place in temporarily and very locally formed solutes in the crystalline material, whereas the overall material remains crystalline. The full reversibility of the solid-state reaction was, furthermore, demonstrated with time-resolved solid-state infrared spectroscopy (observation at the metal-bound SO2 vibration, vs= 1072 cm-1), even after several repeated cycles. Exposure of crystalline samples of 26 alternat-ingly to an atmosphere of SO2 and air did show no loss in signal intensities, e.g. due to the formation of amorphous powder. The release of SO2 from a crystal of 27 was also observed using optical cross-polarization microscopy. A colourless zone (indicative of 26) is growing from the periphery of the crystal whereas the orange colour (indicative for 27) in the core of the crystal diminishes (see Figure 9). 

Shang et al. (61) used microemulsion polymerization to synthesize MWCNT-PMMA composites for gas sensor applications. Better dispersion, enhanced electrical conductivity and better sensor response was observed for in-situ fabricated composites compared to composites prepared by solution mixing. Ma et al. (62) performed in-situ polymerization of MWCNT-PMMA composites in the presence of an AC electric field to study dispersion and alignment of MWCNT in PMMA matrix induced by the electric field. Experimental evidences from in-situ optical microscopy, Raman spectroscopy, SEM and electrical conductivity showed that both dispersion and alignment qualities were significantly enhanced for oxidized MWCNT compared to pristine MWCNT. 

Fig. 1 Scanning electron microscopy (SEM) images showing (a) multiple well pits used to confine the sensor beads (reproduced from [2]). (b) Etched fiber optic end filled with 3 im beads (reproduced from [3]). (c) Etched array housing 2 xm bead (from Illumina )...

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