Cyclic Epipedography─Monitoring Contact Angle Changes in Dipping a Microsphere into a Liquid and Lifting from It.

We propose cyclic epipedography to monitor the wetting characteristics of a microsphere on a stem with a liquid by tracking the level of a liquid line on the sphere when the sphere is dipped into and lifted from the liquid, and therefore the level of the liquid plane at infinity changes with respect to the sphere. Analysis of still images of a video taken with a horizontally held microscope determines the two levels. The sphere allows viewing of the liquid line without being obscured by the meniscus the liquid forms with its container's wall. A nearly perfect sphere is fabricated by melting the tip of a silica optical fiber while being rotated to form a ball 0.5-0.6 mm across. A dip-lift cycle is represented by a cyclic epipedogram─a closed loop in a plot of the level of the liquid line as a function of the level of the liquid plane. The position and shape of the loop allow us to estimate the contact angle at different stages in the dip-lift cycle. We applied the method to a hydroxylated silica microsphere in water, and the same sphere reacted with octylchlorosilane. Then, the method was applied to monitor the progress of the reaction of aminopropyldimethylethoxysilane with surface silanol. We find that the amphiphilic aminopropylsilane makes the surface conform to the environment. In air, the hydrophobic portion comes on top, while in water, hydrophilic part faces the surroundings. This conformity-caused Janus characteristics of the surface were almost absent with the hydroxylated silica and weak with the octylsilane-treated silica.

Whispering Gallery Mode Dip Sensor for Aqueous Sensing.

We report fabrication of a 4 mm thick, preassembled whispering gallery mode (WGM) sensor that can be repeatedly dipped into aqueous solutions and lifted. We built the viable photonic sensor assembly by bending an optical fiber by 90° and molding its tip into a sphere, thus, a long stem holding a submillimeter sensor at the end of a short arm of fiber, and positioning a pair of parallel cone-tipped tapers along the long stem so that the tips touch the sensor. Our sensor head is an optical fiber device just a few millimeters thick and yet has a sensitivity of the resonance wavelength shift comparable to the one obtained with conventional WGM sensors in a planar arrangement. Since dipping and lifting from the solution changes the temperature of the sensor, affecting the resonance wavelength, we enclosed a thermistor within the sensor head to monitor the temperature. We demonstrate that the resonance shift in repeated transfer of the sensor head between water and a solution of sucrose, after correction by the temperature change, is reproducible and agrees with a theoretical estimate of the shift for different concentrations.

All-photonic, dynamic control of optical path length in a silica sphere resonator.

We demonstrate dynamic control of the optical path length for probe light in a spherical dielectric resonator simply by multiplexing intense control light of another color and adjusting its wavelength. The fractional change in the path length, monitored by the resonance wavelengths of whispering gallery modes of the probe light, was nearly equal to the fractional change in the wavelength of the control light. The control was effective in both increasing and decreasing the wavelength, but the weaker the control light or the faster the wavelength change, the narrower the range of control.

Comparison of retention behavior of oligolysine and oligoarginine in ion-pairing chromatography using heptafluorobutyric acid.

This paper describes the retention behavior of oligolysine and oligoarginine peptides of different lengths as a function of heptafluorobutyric acid (HFBA) concentration in ion-pairing reversed-phase chromatography in isocratic elution. A mixture of oligolysine and a mixture of oligoarginine with number of amino acid residues (dp) from two to eight were conveniently prepared by one-pot protease-catalyzed synthesis. Analysis of the logarithm of the retention factor k as a function of [HFBA] for each oligopeptide component, using a closed pairing model, provided values for (1) number (n) of paired HFBA anions per peptide molecule, (2) equilibrium constant (K(ip,m)) for ion pairing between oligopeptides and HFBA anions, and (3) product of the phase ratio and the distribution constant of the paired oligopeptide between the mobile and stationary phases (ßK(d,ip)). We found that ßK(d,ip) of oligoarginine is larger compared with oligolysine having the same dp. A linear relationship was obtained for ln ßK(d,ip) as a function of n + g · dp. By optimizing constant g separately for oligolysine and oligoarginine, we determined that g is larger for oligoarginine, in agreement with the higher hydrophobicity of arginine residues. Plotting the fraction of paired oligoarginine and oligolysine as a function of [HFBA] shows that the cooperative effect in forming ion pairs is greater for oligoarginine than oligolysine.

Light turn-on transient of a whispering gallery mode resonance spectrum in different gas atmospheres.

We examine the resonance spectrum change after turning on the light to feed the fiber taper evanescently coupled to a silica whispering gallery mode (WGM) resonator surrounded by different gases at different pressures. The resonance shifted to a longer wavelength, indicating a temperature rise, before reaching a steady state. The increment was proportional to the power of the light and approximately reciprocally proportional to the thermal conductivity of the surrounding gas, whereas the rate of the shift was approximately proportional to the thermal conductivity. The temperature rise, caused by absorption of intense WGM in silica, was significant even when the wavelength scan range contained only a few tall resonance peaks. We then estimated the power of heat generation and the mean power of WGM during the wavelength scan.

Resonance shifts of transverse-electric whispering gallery modes in a spheroidal resonator.

A variational method was applied to find the wave function of a transverse-electric whispering gallery mode (WGM) in a spheroidal resonator of a uniform refractive index (RI). It was found that the electric field is tangential to the resonator surface as in the sphere, up to the linear order of the ellipticity. Using the wave function, the resonance shift due to adsorption of a thin, uniform dielectric layer onto the surface and the shift by a uniform RI change in the surroundings were evaluated in the perturbation theory. The shift by the RI change is not affected by the ellipticity, but the shift by the layer adsorption now depends on the meridional order. However, the correction is not large unless the ellipticity is large and the meridional order is away from the one for the equatorial mode of WGM.

Removal of dihydroxy-terminated components from monomethoxy-terminated poly(ethylene glycol).

A preparative method to remove dihydroxy-terminated components in a sample of presumably monomethoxy, monohydroxy-terminated poly(ethylene glycol) (PEG) is presented. Purification of the monomethoxy-terminated component allows one to prepare a diblock copolymer of PEG and poly(lactic acid) (PLLA) free of a PLLA-PEG-PLLA triblock copolymer in various biomedical applications of the copolymer. Efficiency of the purification is compared for high osmotic pressure chromatography (HOPC) and preparative size exclusion chromatography (SEC). In HOPC, various types of porous silica particles, surfaces, solvents, polymer concentrations have been screened for the optimal performance. It was found that HOPC is more efficient than SEC, especially HOPC of 30-40 wt% solutions in water by a column packed with acid-washed controlled pore glass is optimal in producing high-purity fractions.

Scale-up study of high osmotic pressure chromatography for separation of poly(epsilon-caprolactone).

Methods to prepare fractions of poly(epsilon-caprolactone) with a narrow molecular mass distribution in large quantities have been examined using high osmotic pressure chromatography under the theta condition. Effects of column dimension and coupling columns in series on the separation resolution were studied. We found that use of a thicker column can improve the resolution if adverse effects of viscous fingering are avoided. We also demonstrated that coupling the columns results in a better separation if the second column does not adsorb high-molecular-mass components purified in the first column.

Composition analysis of poly(ethylene glycol)-poly(L-lactide) diblock copolymer studied by two-dimensional column chromatography.

We used two-dimensional column chromatography to analyze the composition of a sample of presumably a diblock copolymer of poly(ethylene glycol) (PEG) and poly(L-lactide) synthesized from monomethoxy-terminated PEG. The first dimension of the separation is phase fluctuation chromatography to prepare fractions that contain various components of the copolymer in different ratios. The second dimension is size-exclusion chromatography, NMR, and HPLC at the critical condition of PEG. The PEG initiator has small amounts of diol-terminated dimeric components. We found that the copolymer sample contains a triblock copolymer and low-molecular-mass components in addition to the main part of the diblock copolymer. The SEC chromatograms show that the main part consists of two components with distinct peak lengths for the PLLA block. The low-molecular-mass components have a broad distribution in chemical composition. Phase fluctuation chromatography enriched the triblock copolymer and the diblock copolymer with the longer PLLA block in early fractions when the column was packed with carboxymethyl-modified porous silica. When the porous medium was PLLA-grafted silica, size exclusion dominated, but the low-molecular-mass components were separated according to their chemical composition.

Hydrogel formation between enantiomeric B-A-B-type block copolymers of polylactides (PLLA or PDLA: A) and polyoxyethylene (PEG: B); PEG-PLLA-PEG and PEG-PDLA-PEG.

A mixed suspension of the enantiomeric B-A-B triblock copolymers, polyoxyethylene-block-poly(L-lactide)-block-polyoxyethylene (PEG-PLLA-PEG) and polyoxyethylene-block-poly(D-lactide)-block-polyoxyethylene (PEG-PDLA-PEG), was found to induce reversible gel-to-sol transition depending on the polymer concentration and temperature. The storage and loss moduli of the gel formed at lower temperature were much higher than those of the gel prepared from the corresponding ABA-type triblock copolymers because of the higher polymer concentration in the former. Although the stereo-complexation of the PLLA and PDLA blocks occurred at higher temperature also in the B-A-B copolymers, it was not responsible for the gelation of the mixed suspension. The PEG chains, involved in the helix formation of the PLLA and PDLA, should form helices with opposite helical senses to aggregate and lead the gelation of the system.

Reversed phase ion-pairing chromatography of an oligolysine mixture in different mobile phases: effort of searching critical chromatography conditions.

Our earlier study [J. Chromatogr. A 1218 (2011) 7765] on separation of an oligolysine mixture consisting of chains with 2-8 lysine residues (number of lysine residues, dp=2-8) by ion-pairing reversed-phase chromatography using heptafluorobutyric acid (HFBA) as an ion pairing reagent at fixed mobile phase acetonitrile (ACN) content was extended to isocratic elution conditions with different ACN percentages. The present work explored how manipulating the mobile phase HFBA concentration ([HFBA]) and %-ACN content influences separations of the oligolysine mixture. The closed pairing model was used to analyze variation of the retention factor as a function of [HFBA]. The partition coefficient of the paired peptide decreased with increasing %-ACN. Pairing of HFBA to oligolysine was cooperative, and the effect increased when %-ACN in the mobile phase was lowered. A plot of the partition coefficient as a function of %-ACN for oligolysines varying in dp converged at one ACN content, indicating a critical condition in which components of different dp co-elute.

Cooperative effect in ion pairing of oligolysine with heptafluorobutyric acid in reversed-phase chromatography.

The retention behavior of an oligolysine mixture, consisting of two to eight residues, was examined at different concentrations of heptafluorobutyric acid (HFBA) in the mobile phase using a C18 column. A single ion record (SIR) mode of the mass spectrometer produced a distinct retention time for each oligomer component. As the concentration of HFBA increased, the retention time of each oligomer increased. Furthermore, the increase in retention time is chain-length dependent such that, the longer the oligomer chain, the more rapid was the rate that retention time increased. A closed pairing model that presumes an equilibrium between the unpaired state and the paired state with a fixed number of HFBA molecules was used to analyze the retention factor as a function of [HFBA]. Curve fitting gave estimates of the ion-pairing equilibrium constant (K(ip,m)), the distribution constant of paired oligolysine (K(D,ip)), and the number of paired HFBA for each oligolysine (n). The plot of the fraction of paired oligolysine in the mobile phase, estimated from K(ip,m) and n as a function of [HFBA], revealed a cooperative effect. In contrast, an open pairing model that assumes independent pairing of HFBA with each residue failed to describe the observed retention behavior.

Coupled whispering gallery modes in a multilayer-coated microsphere.

Whispering gallery modes in a microsphere coated with three layers of high, low, and high refractive indices (RIs) are considered. Coastal modes and inland modes, centered on the outer and inner high-RI layers, exist as different radial modes. At right values of RI and thickness of the three layers, an inland mode and a coastal mode couple to produce a radial distribution of the photonic field that resembles bonding and antibonding orbitals. The coupling occurs across a layer of the middle low-RI layer, much thicker than the wavelength of light. The coupling is analyzed in a quantum-mechanical analog of a one-dimensional particle in a double-well potential.

Detection of protein orientation on the silica microsphere surface using transverse electric/transverse magnetic whispering gallery modes.

The state of adsorbed protein molecules can be examined by comparing the shifts in a narrow line resonance wavelength of transverse electric (TE) and transverse magnetic (TM) whispering gallery modes (WGM) when the molecules adsorb onto a transparent microsphere that houses WGM. In adsorption of bovine serum albumin (BSA) onto an aminopropyl-modified silica microsphere, the TM/TE shift ratio indicated highly anisotropic polarizability of BSA in the direction normal to the surface, most likely ascribed to anchoring the heart-shaped protein molecule by one of its tips. The polarization-dependent resonance shift was confirmed when the surrounding refractive index was uniformly changed by adding salt, which would simulate adsorption of large objects.

Nanolayer characterization through wavelength multiplexing of a microsphere resonator.

We optically characterize nanolayer (<150 nm) formation in situ on a silica microsphere in an aqueous environment by simultaneously following the shifts of whispering-gallery modes at two wavelengths. This approach was inspired by layer perturbation theory, which indicates that these two measurements can be used to determine independently both the thickness and the optical dielectric constant. The theory is verified for extreme cases and used to characterize a biophysically relevant hydrogel nanolayer with an extremely small excess refractive index of 0.0012.

Multiplexed DNA quantification by spectroscopic shift of two microsphere cavities.

We have developed a novel, spectroscopic technique for high-sensitivity, label-free DNA quantification. We demonstrate that an optical resonance (whispering gallery mode) excited in a micron-sized silica sphere can be used to detect and measure nucleic acids. The surface of the silica sphere is chemically modified with oligonucleotides. We show that hybridization to the target DNA leads to a red shift of the optical resonance wavelength. The sensitivity of this resonant technique is measured as 6 pg/mm(2) mass loading, higher as compared to most optical single-pass devices such as surface plasmon resonance biosensors. Furthermore, we show that each microsphere can be identified by its unique resonance wavelength. Specific, multiplexed DNA detection is demonstrated by using two microspheres. The multiplexed signal from two microspheres allows us to discriminate a single nucleotide mismatch in an 11-mer oligonucleotide with a high signal-to-noise ratio of 54. This all-photonic whispering gallery mode biosensor can be integrated on a semiconductor chip that makes it an easy to manufacture, analytic component for a portable, robust lab-on-a-chip device.