Concentration dependence of fluorescence signal in a microfluidic fluorescence detector

Fluorescence detection is one of the most widely used techniques for measuring analyte concentration in biological applications. Although used frequently with bulk systems, there are important considerations in using this approach in thin (<100 μm) microfluidic lab-on-a-chip (LOC) systems in relating the measured fluorescence signal to analyte concentration. In this work, a general relationship between the photoresponse of an on-chip organic photodetector (OPD) and concentration of a fluorescent dye Rhodamine 6G in a microfluidic chip is presented. The developed theoretical model for photoresponse matches well to the extreme sub-linearity observed in measurements in a dilution series from 10 nM to 1 mM. The influence of the system factors, such as noise, detector sensitivity, and optical power were also analyzed to indicate design changes to improve the system performance and limits of detection even further.     

Design and fabrication of long focal length microlens arrays

In this paper, we present microlens arrays (MLA) with long focal length (in millimeter range) based on thermal reflow process. The focal length of microlens is usually in the same order of lens diameter or several hundred microns. To extend focal length, we made a photoresist (SU-8) MLA covered by a Polydimethylsiloxane (PDMS) film on a glass substrate. Because the refractive index difference between PDMS and photoresist interface is lower than that of air and MLA interface, light is less bended when passing through MLA and is focused at longer distance. Microlenses of diameters from 50 μm to 240 μm were successfully fabricated. The longest focal length was 2.1 mm from the microlens of 240 μm diameter. The numerical aperture (NA) was reduced 0.06, which is much lower than the smallest NA (~ 0.15) by regular thermal reflow processes. Cured PDMS has high transmittance and becomes parts of MLA without too much optical power loss. Besides, other focal lengths can be realized by modifying the refractive index different between two adjacent materials as described in this paper.     

Aberration athermal design for infrared search and trace optical system

To satisfy environment requirement of infrared search and trace optical system, an infrared diffractive/refractive hybrid optical system in 3.7–4.8 μm with 11.42° of field of view for passive athermalization is presented. The system is consisted of three lenses, including two aspheric surfaces and a diffraction surface, which has only two materials Ge and Si. The optical system has compact structure, small volume and light weight. The image quality of the system approaches the diffraction limit in the temperature range −80 °C to 160 °C. It is compatible with staring focal plane array which has a format of 320 × 240 and the pixel pitch of 30 μm. The system need not move the compensated lens repeatedly to obtain the best images from −80 °C to 160 °C and enhances the performance of target tracking and recognition.     

Optical design of a full-pupil field,non-contact and high resolution corneal curved objective lens

It has been a challenge to overcome the corneal curvature radius to design a full-pupil field, non-contact and high resolution corneal curved objective lens, which covers the cornea full-pupil field and has the ability to resolve corneal cells. In this paper, we report an optical design of a full-pupil field, non-contact corneal curved objective lens for high resolution cornea imaging. The advantages of this lens are that it has a wide field of view (FOV) with the corneal curved image surface, maintains the beam normal incidence, as well as non-contact lens imaging, and offers a cell-level lateral resolution of cornea structure. The analysis of optimization shows that the system achieves diffraction limit in a circular FOV of 4 mm diameter covering the full-pupil zone. The theoretical lateral resolution is about 2.5 μm with an image space NA of 0.16, which is sufficient to resolve corneal cells of 7 μm diameter, and the working distance is larger than 15 mm which is enough for a non-contact objective lens. So the optical design is effectively and efficiently meeting the demand of specifications.     

The nonlinear refraction and absorption dependence on the thermal effect for 4 ns pulse duration in binuclear Zn(II) phthalocyanine solution

The influence of thermal effect on the third-order nonlinear optical properties of binuclear Zn(II) phthalocyanine in chloroform solution was studied. The nonlinear refraction and absorption of the sample was measured by using Z-scan technique with 4 ns laser pulses at 532 nm wavelength. The opposite signs of the effective nonlinear refraction index were observed by changing the focal length of focusing lens from 10 cm to 20 cm in the experimental setup. Changing the focusing lens increased the beam waist radius from 7 μm to 20 μm. The nonlinear absorption coefficient was reduced about 200 times based on changing the fluence or beam waist radius. The drastic changes in the third-order nonlinear optical parameters were attributed to thermal effect. To investigate the role of thermal effects even further the effective nonlinear refraction and absorption coefficients were studied by using different repetition rates, input powers and concentrations.     

Demonstration of a no-moving-parts axial scanning confocal microscope using liquid crystal optics

For the first time, to the best of authors’ knowledge, a no-moving-parts axial scanning confocal microscope (ASCM) system is designed and demonstrated using a combination of a large diameter liquid crystal (LC) lens and a classical microscope objective lens. By electrically controlling the 5 mm diameter LC lens, the 633 nm wavelength focal spot is moved continuously over a 48 μm range with a measured 3-dB axial resolution of 3.1 μm using a 0.65 numerical aperture (NA) micro-objective lens. The ASCM is successfully used to image an Indium Phosphide (InP) twin square optical waveguide sample with a 10.2 μm waveguide pitch and 2.3 μm height and width. Using fine analog electrical control of the LC lens, a super-fine sub-wavelength axial resolution of 270 nm is demonstrated. The proposed ASCM can be useful in various precision three-dimensional (3D) imaging and profiling applications.     

Characterization of ultra-weak fluorescence using picosecond non-collinear optical parametric amplifier

We report a technique for characterization of ultra-weak fluorescence based on a 355-nm pumped picosecond non-collinear optical parametric amplifier (OPA). In the experiment, we effectively reduced the strong super-fluorescence background by using a series of methods. With the picosecond OPA as the pre-amplifier and the gating pulse, the decay of the fluorescence of Rhodamine 6G dye in ethanol was measured and the fluorescence lifetime was found to be about 941 ps. The gain factor of this parametric fluorescence amplifier was measured to be ∼4.2 × 10⁶, while the energy detection limit was ∼160 aJ per pulse within a 15-ps gating pulse.     

Design of midwave infrared athermalization optical system with a large focal plane array

Based on the most advanced staring focal plane array which had a format of 640 × 480 and the pixel pitch of 15 μm, a set of all-sphere midwave infrared ahermalization optical system was designed. The working wavelength was in 3–5 μm, the full field of view was 8.58°, the relative aperture was 1/2, the efficient focal length (EFL) was 80 m. The opticalsystem consisted of four lenses with three kinds of material – Ge, ZnSe and Si. All surfaces were sphere, which was easier to process test, making the cost inexpensive, and it could avoid using diffractive surface and aspheric surface. The image quality of the system approaches the diffraction limit in the temperature range −60 °C-180 °C. The design results proved that, the high resolution midwave infrared optical system had compact structure, small volume, high resolution and excellent image quality, meeting the design requirements, so that it could be used for photoelectric detection and tracking system.     

Digital Micro-mirror Device-based broadband optical image sensor for robust imaging applications

To the best of the authors' knowledge, presented for the first time is the design of a robust broadband optical image sensor using a Digital Micro-mirror Device (DMD). Electronic focus control of the imaging lens and full programmability of the spatial sampling aperture shape, size, and location on the DMD plane that mechanically scans the incident incoherent optical irradiance distribution lead to imaging smartness. Dual port single-point photo-detection design provides imaging operation robustness to the global light irradiance variations such as via environmental effects, e.g., moving clouds. As the Texas Instruments (TI) DMD can provide light modulation over 400 nm to 2500 nm wavelengths, visible, Near Infrared (NIR), and Short-Wave Infrared (SWIR) bands can be simultaneously processed to generate three independent band images via three point photo-detectors. A proof-of-concept experiment in the SWIR band at 1580 nm is conducted using an incoherent heart-shaped target that is sampled using the DMD imager set for a 68.4 μm side square moving pinhole. A 60 × 60 pixel image from the proposed imager produces a 0.94 cross-correlation peak when compared to an optically attenuated heart shape image produced by a near 9 μm pixel size phosphor coated Charge Coupled Device (CCD) imager. Using the dual-detection method, robust 633 nm visible light imaging of an Air Force (AF) Chart figure is successfully demonstrated for 3 Hz global light fluctuation. Applications for the proposed imager include optical sensing in the fields of astronomy, defense, medicine, and security.     

Fabrication of Si-based waveguide splitters using photosensitive sol-gel method

In this paper, a photosensitive sol-gel method for 1 × 8 Y-branch optical splitter is reported. The poly (methyl methacrylate) (PMMA) cladding films with different thicknesses (above 10 μm) on Si substrates are prepared on Si substrates by dip-coating, the root-mean-square roughness (Rrms) of which is less than 0.7 nm. The UV photosensitive organic-inorganic composite SiO₂/ZrO₂/H core films are then coated on these PMMA cladding films using photosensitive sol-gel method. The refractive indexes of PMMA cladding films examined by spectroscopic ellipsometer are 1.4854, 1.4815 and 1.4806 at 0.85, 1.31 and 1.55 μm, while that of the SiO₂/ZrO₂/H gel films are 1.5569, 1.5489 and 1.5472 μm, respectively, which is larger than that of PMMA cladding films. Therefore, the composite structure of SiO₂/ZrO₂/H gel films on PMMA-on-Si substrates could be used to fabricate waveguide splitter. Based on the inherent photosensitivity of the SiO₂/ZrO₂/H gel film, 1 × 8 Y-branch optical power splitters with a thickness of 5 μm are fabricated by irradiating the gel film with UV light through a mask followed by dissolving the non-irradiated area in a suitable solvent. The line-width and output spacing of this splitter are 25 and 110 μm, respectively. The observed near-field pattern indicates that the light with a wavelength of 1.53-1.56 μm can be transmitted and split into eight branches in the optical splitter, which is fabricated by using the above technique.     

Study on the interaction of tropisetron hydrochloride and l-tryptophan by spectrofluorimetry and its analytical application

A new method to determine tropisetron hydrochloride with l-tryptophan in the medium with pH=9.0 was studied, which is based on the fluorescence quenching effect of tropisetron hydrochloride on l-tryptophan. The fluorescence quenching mechanism and various factors influencing fluorescence quenching were discussed. Under the optimum conditions, the linear range and detection limit were 0.03-12.0 and 0.01 μg/mL (correlation coefficient r=0.9970), respectively. The calibration curve equation was ΔF=6.17+12.56 C (μg/mL). RSD was 3.4% (c=4.0 μg/mL, n=5); the detection limit estimated (S/N=3) was 0.01 μg/mL. The proposed method had been successfully applied to determine tropisetron hydrochloride in real samples and the obtained results were in good agreement with the results of the official method.     

Two-dimensional optical micro-scanner on silicon technology

A two-dimensional optical micro-scanner, which main components are two mobile flat and a concave micro-mirrors, is designed such that, all optical components can be fabricated on the same substratum. The optical parameters, which physical dimensions are between 50 and 500 μm, are obtained within the geometrical optics. The optical performance is evaluated by means of the MTF and Rayleigh resolution criteria, given 80% of modulation for a frequency of 8 cycles/mm with a Gaussian source, the resolution limit is 30 μm.     

Femtosecond laser embedded grating micromachining of flexible PDMS plates

We report on the femtosecond laser micromachining of photo-induced embedded diffraction grating in flexible Poly (Dimethly Siloxane) (PDMS) plates using a high-intensity femtosecond (130 fs) Ti: sapphire laser (λ_p = 800 nm). The refractive index modifications with diameters ranging from 2 μm to 5 μm were photo-induced after the irradiation with peak intensities of more than 1 × 10¹¹ W/cm². The graded refractive index profile was fabricated to be a symmetric around from the center of the point at which femtosecond laser was focused. The maximum refractive index change (Δn) was estimated to be 2 × 10⁻³. By the X-Y-Z scanning of sample, the embedded diffraction grating in PDMS plate was fabricated successfully using a femtosecond laser.     

Optimization of spin-valve parameters for magnetic bead trapping and manipulation

Magneto-optic Kerr effect (MOKE) and magnetoresistance (MR) measurements were used to measure the switching characteristics of spin-valve (SV) arrays currently being developed to trap and release superparamagnetic beads within a fluid medium. The effect of SV size on switching observed by MOKE showed that a 1 μm×8 μm SV element was found to have optimal switching characteristics. MR measurements on a single 1 μm×8 μm SV switched with either an external applied magnetic field or a local magnetic field generated by an integrated write wire (current density ranging from 10⁶ to 10⁷ A/cm²) confirmed the MOKE findings. The 1 μm×8 μm SV low field switching was observed to be +8 and −2 mT with two stable states at zero field; the high field switching was observed to be −18 mT. The low switching fields and the large magnetic moment of the SV trap along with our observation of minimal magnetostatic effects for dense arrays are necessary design characteristics for high-force, “switchable-magnet,” microfluidic bead trap applications.     

Design of organic 3D microresonators with microfluidics coupled to thin-film processes for photonic applications

We report on the design and realization of photonic integrated devices based on 3D organic microresonators (MR) shaped by an applied fluid mechanism technique. Such an interdisciplinary approach has been judiciously achieved by combining microfluidics techniques and thin-film processes, respectively, for the realizations of microfluidic and optical chips. The microfluidic framework with flow rates control allows the fabrication of microresonators with diameters ranging from 30 to 160 μm. The resonance of an isolated sphere in air has been demonstrated by way of a modified Raman spectroscopy devoted to the excitation of Whispering Gallery Modes (WGM). Then the 3D-MR have been integrated onto an organic chip and positioned either close to the extremity of a taper or alongside a rib waveguide. Both devices have proved efficient evanescent coupling mechanisms leading to the excitation of the WGM confined at the surface of the organic 3D-MR. Finally, a band-stop filter has been used to detect the resonance spectra of organic resonators once being integrated. Such spectral resonances have been observed with an integrated configuration and characterized with a Δλ = 1.4 nm free spectral range (FSR), appearing as stemming from a 78 μm-radius MR structure.     

Fluorescence enhancement effect for the determination of curcumin with yttrium(III)—curcumin—sodium dodecyl benzene sulfonate system

It is found that the fluorescence of curcumin is greatly enhanced by yttrium(III) (Y³⁺) in the presence of sodium dodecyl benzene sulfonate. Based on this, a sensitive fluorimetric method for the determination of curcumin in aqueous solution is proposed. In the potassium hydrogen phthalate (KHP) buffer, the fluorescence intensity of curcumin is proportional to the concentration of curcumin in the range of 7.37×10⁻⁴-0.18, 0.18-2.95 μg mL⁻¹ and the detection limit is 0.1583 ng mL⁻¹. The actual samples are satisfactorily determined. In addition, the interaction mechanism is also studied.     

An optical technique for remote focusing in microscopy

We describe the theory of a new method of optical refocusing that is particularly relevant for confocal and multiphoton microscopy systems. This method avoids the spherical aberration that is common to other optical refocusing systems. We show that aberration-free refocusing can be achieved over an axial scan range of 70 μm for a 1.4 NA objective lens. As refocusing is implemented remotely from the specimen, this method enables high axial scan speeds without mechanical interference between the objective lens and the specimen.     

High repetition rate mid-infrared generation with singly resonant optical parametric oscillator using multi-grating periodically poled MgO:LiNbO3

A high repetition rate mid-infrared singly resonant optical parametric oscillator (OPO) using MgO-doped multi-grating periodically poled LiNbO₃ (MgO:PPLN) is demonstrated. A 1064 nm Q-switched Nd:YVO₄ laser at 10 kHz repetition rate and pulse width of 17.8 ns was used to pump the OPO. The period of the quasi-phase matched (QPM) grating in the multi-grating MgO:PPLN chip varied from 25.5 to 31.5 μm in steps of 0.5 μm. This corresponds to the generation of a signal beam from 1.37 to 1.64 μm and an idler beam from 3.0 to 4.8 μm, respectively. A maximum signal power of 250 mW and idler power of 140 mW has been obtained with an input pump beam of power 1.92 W, for a grating period of 30.5 μm. A maximum optic-optic conversion efficiency of 20% and 7.4% in the idler has been observed. It has been observed that the output power increases as the period of the grating increases.     

Quadrature fringes wide-field optical coherence tomography and its applications to biological tissues

We propose and demonstrate quadrature fringes wide-field optical coherence tomography (QF WF OCT) to expand an optical Hilbert transformation to two-dimensions. This OCT simultaneously measures two quadrature interference images using a single InGaAs CCD camera to obtain en face OCT images. The axial and lateral resolutions are measured at 29 μm in air and 70 μm limited by a pixel size of camera using a superluminescent diode with a wavelength of 1.3 μm as the light source; the system sensitivity is determined to be −90 dB. The area of the en face OCT images is 4.0 mm × 4.0 mm (160 × 160  pixels). The OCT images are measured axially with steps of 10 μm. The en face OCT images of a in vivo human fingertip and a in situ rat brain are three-dimensionally measured up to the depth of about 3 mm with some degradations of a lateral resolution.     

Surface modification of poly(dimethylsiloxane) for microfluidic assay applications

The surface of a poly(dimethylsiloxane) (PDMS) film was imparted with patterned functionalities at the micron-scale level. Arrays of circles with diameters of 180 and 230 μm were functionalized using plasma oxidation coupled with aluminum deposition, followed by silanization with solutions of 3-aminopropyltrimethoxy silane (3-APTMS) and 3-mercaptopropyltrimethoxy silane (3-MPTMS), to obtain patterned amine and thiol functionalities, respectively. The modification of the samples was confirmed using X-ray photoelectron spectroscopy (XPS), gold nanoparticle adhesion coupled with optical microscopy, as well as by derivatization with fluorescent dyes. To further exploit the novel surface chemistry of the modified PDMS, samples with surface amine functionalities were used to develop a protein assay as well as an array capable of cellular capture and patterning. The modified substrate was shown to successfully selectively immobilize fluorescently labeled immunoglobulin G (IgG) by tethering Protein A to the surface, and, for the cellular arrays, C2C12 rat endothelial cells were captured. Finally, this novel method of patterning chemical functionalities onto PDMS has been incorporated into microfluidic channels. Finally, we demonstrate the in situ chemical modification of the protected PDMS oxidized surface within a microfluidic device. This emphasizes the potential of our method for applications involving micron-scale assays since the aluminum protective layer permits to functionalize the oxidized PDMS surface several weeks after plasma treatment simply after etching away the metallic thin film.