Browsing by Author "Hee, Ping-Yu"

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In situ laser micropatterning of proteins for dynamically arranging living cells
(2013)
Okano, Kazunori
;
Matsui, Ai
;
Maezawa, Yasuyo
;
Hee, Ping-Yu
;
Matsubara, Mie
This study shows the modification of the surface of polymer-layered glass substrates to form biofunctional micropatterns through femtosecond laser ablation in an aqueous solution. Domains of micrometer size on a substrate can be selectively converted from proteinphobic (resistant to protein adsorption) to proteinphilic, allowing patterning of protein features under physiological aqueous conditions. When femtosecond laser pulses (800 nm, 1 kHz, 200-500 nJ per pulse) were focused on and scanned on the substrate, which was glass covered with the proteinphobic polymer 2-methacryloyloxyethylphosphorylcholine (MPC), the surface became proteinphilic. Surface analysis by X-ray photoelectron spectroscopy (XPS) and scanning electron microscopy (SEM) reveals that the laser ablates the MPC polymer. Extracellular matrix (ECM) proteins were bound to the laser-ablated surface by physisorption. Since femtosecond laser ablation is induced under physiological aqueous conditions, this approach can form micropatterns of functional ECM proteins with minimal damage. This method was applied to pattern collagen, laminin, and gelatin on the substrate. Removal of an ECM protein from the substrate followed by replacement with another ECM protein was achieved on demand at a specific location and time by the same laser ablation method. Living cells adhered to the fabricated domains where ECM proteins were arranged. The modification of patterning during cell culture was used to control cell migration and form arrays of different cells.
Rayleigh scattering correlation spectroscopy on diffusion dynamics of nanoparticles under intense laser irradiation
(2013)
Hee, Ping-Yu
;
Uwada, Takayuki
;
Okano, Kazunori
;
Miura, Atsushi
;
Masuhara, Hiroshi
Rayleigh scattering correlation microspectroscopy is developed and applied to study diffusion dynamics of some nanospheres in water. It was clearly found that the diffusion constant of gold nanoparticles decreased with increasing excitation laser power at the excitation wavelength of higher absorption cross section. This behavior was explained in terms of a coupling between laser trapping by the scattering excitation laser itself and laser heating of the particle. In the case of non-absorbing nanospheres such as silica and polystyrene, the excitation power dependence can be ascribed only to the laser trapping. Experimental setup is introduced, theoretical formulation is described, and future development of this measurement is considered.
Size-Dependent Optical Properties of Grana Inside Chloroplast of Plant Cells
(2017-02-09)
Uwada, Takayuki
;
Huang, Ling-Ting
;
Hee, Ping-Yu
;
Usman, Anwar
;
Masuhara, Hiroshi
Well-packed thylakoids known as grana are one of the major functional sites for photosynthesis in algae and plants. Their highly ordered structures can be considered as a few hundred nanometer-sized particles having distinct scattering cross sections from other various macromolecular organizations inside plant cells. With this background we shOw that elastic light scattering imaging and microspectroscopy is an important tool for investigating structure and organisation of grana inside a single chloroplast in plant tells. We have demonstrated this noninvasive method to identify the distribution of grana in intact fresh leaf of robust and rapidly growing Egaria densa, which is also known as Anachris and among the most popular aquarium plants. The scattering efficiency spectra of their individual grana fairly resemble cooperative absorption spectra of porphyrins and carotenoids. We found that the electronic structure of the stacked thylakoids shows granum size-dependence, indicating that size of grana is one of the critical parameters in the regulation of the photochemical functions in the thylakoid.