Arthur Chiou

Size-dependent endocytosis of gold nanoparticles studied by three-dimensional mapping of plasmonic scattering images

BackgroundUnderstanding the endocytosis process of gold nanoparticles (AuNPs) is important for the drug delivery and photodynamic therapy applications. The endocytosis in living cells is usually studied by fluorescent microscopy. The fluorescent labeling suffers from photobleaching. Besides, quantitative estimation of the cellular uptake is not easy. In this paper, the size-dependent endocytosis of AuNPs was investigated by using plasmonic scattering images without any labeling.ResultsThe scattering images of AuNPs and the vesicles were mapped by using an optical sectioning microscopy with dark-field illumination. AuNPs have large optical scatterings at 550-600 nm wavelengths due to localized surface plasmon resonances. Using an enhanced contrast between yellow and blue CCD images, AuNPs can be well distinguished from cellular organelles. The tracking of AuNPs coated with aptamers for surface mucin glycoprotein shows that AuNPs attached to extracellular matrix and moved towards center of the cell. Most 75-nm-AuNPs moved to the top of cells, while many 45-nm-AuNPs entered cells through endocytosis and accumulated in endocytic vesicles. The amounts of cellular uptake decreased with the increase of particle size.ConclusionsWe quantitatively studied the endocytosis of AuNPs with different sizes in various cancer cells. The plasmonic scattering images confirm the size-dependent endocytosis of AuNPs. The 45-nm-AuNP is better for drug delivery due to its higher uptake rate. On the other hand, large AuNPs are immobilized on the cell membrane. They can be used to reconstruct the cell morphology.

A comparative study of living cell micromechanical properties by oscillatory optical tweezers

Micromechanical properties of biological cells are crucial for cells functions. Despite extensive study by a variety of approaches, an understanding of the subject remains elusive. We conducted a comparative study of the micromechanical properties of cultured alveolar epithelial cells with an oscillatory optical tweezer-based cytorheometer. In this study, the frequency-dependent viscoelasticity of these cells was measured by optical trapping and forced oscillation of either a submicron endogenous intracellular organelle (intra-cellular) or a 1.5microm silica bead attached to the cytoskeleton through trans-membrane integrin receptors (extra-cellular). Both the storage modulus and the magnitude of the complex shear modulus followed weak power-law dependence with frequency. These data are comparable to data obtained by other measurement techniques. The exponents of power-law dependence of the data from the intra- and extra- cellular measurements are similar; however, the differences in the magnitudes of the moduli from the two measurements are statistically significant.

Shift-tolerance property of an optical double-random phase-encoding encryption system

We investigate the shift-tolerance property of the decrypting phase mask in an optical double-random phase-encoding encryption system. A simple method for improving the shift tolerance of the phase mask is proposed. We demonstrate how the robustness to data loss of the encrypted image extends the shift tolerance of the decrypting phase mask. The signal-to-noise ratio is calculated. Both a computer simulation and an experiment are presented.

One-dimensional jumping optical tweezers for optical stretching of bi-concave human red blood cells

We report the experimental demonstration of optical stretching of individual bio-concave human red blood cells (RBCs) with one-dimensional jumping optical tweezers. We trapped a RBC in isotonic buffer solution in a conventional stationary single-beam gradient-force optical trap and discretely scanned the trapping beam with an acousto-optic modulator such that the focal point of the trapping beam jumped back-and-forth between two fixed points. At the jumping frequency on the order of a 100 Hz and higher, and the jumping distance in the range of a few microns, the bi-concave RBC was stably trapped and stretched. The elongation of the stretched RBC was measured as a function of the beam-scanning amplitude, and the experimental results were explained qualitatively by a theoretical model.

Dynamic deformation of red blood cell in Dual-trap Optical Tweezers

Three-dimensional dynamic deformation of a red blood cell in a dual-trap optical tweezers is computed with the elastic membrane theory and is compared with the experimental results. When a soft particle is trapped by a laser beam, the particle is deformed depending on the radiation stress distribution whereas the stress distribution on the particle in turn depends on the deformation of its morphological shape. We compute the stress re-distribution on the deformed cell and its subsequent deformations recursively until a final equilibrium state solution is achieved. The experiment is done with the red blood cells in suspension swollen to spherical shape. The cell membrane elasticity coefficient is obtained by fitting the theoretical prediction with the experimental data. This approach allows us to evaluate up to 20% deformation of cells shape.

Local scattering stress distribution on surface of a spherical cell in optical stretcher

We calculate stress distribution on the surface of a spherical cell trapped by two counter propagating beams in the optical stretcher in the ray optics regime. We demonstrate that the local scattering stress is perpendicular to the spherical refractive surface regardless of incident angle, polarization and the reflectance and transmittance at the surface. We explain the apparition of peaks in the stress distribution, which were not revealed in the existing theory. We consider the divergence of the incident beams from the fibers, and express the stress distribution as a function of fiber-to-cell distance. The new theory can predict the cells deformation more precisely.

Three-dimensional optical force field on a Chinese hamster ovary cell in a fiber-optical dual-beam trap

We used a fiber-optical dual-beam trap (single-mode fiber, lambda = 532nm, trapping power ~ 22mW, the distance between the two fiber end-faces = 125mum) to capture a Chinese hamster ovary (CHO) cell with a diameter of approximately 15mum and tracked its three-dimensional Brownian motion via a pair of orthogonal quadrant photodiodes. By analyzing the Brownian motion of the trapped CHO cell, we determined the force constants of the optical force field on the CHO cell to be k(x)=6.75 pN/mum, k(y)=5.53 pN/mum, k(z)=1.96 pN/mum, and k(x)=2.91 pN/mum, k(y)=2.7 pN/mum, k(z)=0.79 pN/mum, respectively, before and after the CHO cell was treated with latrunculin, a toxic drug known to disrupt the cytoskeleton of the cell.

Lateral shifting sensitivity of a ground glass for holographic encryption and multiplexing using phase conjugate readout algorithm

An encryption-selectable holographic storage algorithm with use of random phase encoding and angular multiplexing is proposed and demonstrated. The pattern for storage can be encrypted and multiplexed with use of a ground glass, which is also used to decrypt the pattern. A phase conjugation of the reference beam is used to generate phase conjugate waves of the stored patterns. The lateral shifting sensitivity of the ground glass used to decrypt the information is theoretically analyzed and is compared with experimental measurement. The shifting tolerance for various ground glasses under imperfection phase reconstruction in the decryption processing is studied.

Optofluidic platform for real-time monitoring of live cell secretory activities using Fano resonance in gold nanoslits.

An optofluidic platform for real-time monitoring of live cell secretory activities is constructed via Fano resonance in a gold nanoslit array. Large-area and highly sensitive gold nanoslits with a period of 500 nm are fabricated on polycarbonate films using the thermal-annealed template-stripping method. The coupling between gap plasmon resonance in the slits and surface plasmon polariton Bloch waves forms a sharp Fano resonance with intensity sensitivity greater than 11 000% per refractive index unit. The nanoslit array is integrated with a cell-trapping microfluidic device to monitor dynamic secretion of matrix metalloproteinase 9 (MMP-9) from human acute monocytic leukemia cells in situ. Upon continuous lipopolysaccharide (LPS) stimulation, MMP-9 secretion is detected within 2 h due to ultrahigh surface sensitivity and close proximity of the sensor to the target cells. In addition to the advantage of detecting early cell responses, the sensor also allows interrogation of cell secretion dynamics. Furthermore, the average secretion per cell measured using our system well matches previous reports while it requires orders of magnitude less cells. The optofluidic platform may find applications in fundamental studies of cell functions and diagnostics based on secretion signals.

Three-dimensional tracking of Brownian motion of a particle trapped in optical tweezers with a pair of orthogonal tracking beams and the determination of the associated optical force constants

We report the first experimental results on quantitative mapping of three-dimensional optical force field on a silica micro-particle and on a Chinese hamster ovary cell trapped in optical tweezers by using a pair of orthogonal laser beams in conjunction with two quadrant photo-diodes to track the particles (or the cells) trajectory, analyze its Brownian motion, and calculate the optical force constants in a three-dimensional parabolic potential model. For optical tweezers with a 60x objective lens (NA = 0.85), a trapping beam wavelength lambda = 532nm, and a trapping optical power of 75mW, the optical force constants along the axial and the transverse directions (of the trapping beam) were measured to be approximately 1.1x10-8N/m and 1.3x10-7N/m, respectively, for a silica particle (diameter = 2.58microm), and 3.1x10-8 N/m and 2.3x10-7 N/m, respectively, for a Chinese hamster ovary cell (diameter ~ 10 microm to 15microm). The set of force constants (Kx, Ky, and Kz ) completely defines the optical force field E(x, y, z) = [Kx x2 + Ky y2 + Kz z2]/2 (in the parabolic potential approximation) on the trapped particle. Practical advantages and limitations of using a pair of orthogonal tracking beams are discussed.