• Journal of the Korean Society for Precision Engineering
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• v.22 no.4
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• pp.84-91
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• 2005

We measured the pitch of one-dimensional (ID) grating specimens using a metrological atomic force microscope (M-AFM). The ID grating specimens a.e often used as a magnification standard in nano-metrology, such as scanning probe microscopy (SPM) and scanning electron microscopy (SEM). Thus, we need to certify the pitch of grating specimens fur the meter-traceability in nano-metrology. To this end, an M-AFM was setup at KRISS. The M-AFM consists of a commercial AFM head module, a two-axis flexure hinge type nanoscanner with built-in capacitive sensors, and a two-axis heterodyne interferometer to establish the meter-traceability directly. Two kinds of ID grating specimens, each with the nominal pitch of 288 nm and 700 nm, were measured. The uncertainty in pitch measurement was evaluated according to Guide to the Expression of Uncertainty in Measurement. The pitch was calculated from 9 line scan profiles obtained at different positions with 100 ㎛ scan range. The expanded uncertainties (k = 2) in pitch measurement were 0.10 nm and 0.30 nm for the specimens with the nominal pitch of 288 nm and 700 nm. The measured pitch values were compared with those obtained using an optical diffractometer, and agreed within the range of the expanded uncertainty of pitch measurement. We also discussed the effect of averaging in the measurement of mean pitch using M-AFM and main components of uncertainty.

• Journal of the Optical Society of Korea
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• v.18 no.5
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• pp.551-557
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• 2014

In this study, we demonstrated multimodal nonlinear optical (NLO) microscopy integrated simultaneously with two-photon excitation fluorescence (TPEF), second-harmonic generation (SHG), and coherent anti-Stokes Raman scattering (CARS) in order to obtain targeted cellular and label-free images in an immunofluorescence assay of the atherosclerotic aorta from apolipoprotein E-deficient mice. The multimodal NLO microscope used two laser systems: picosecond (ps) and femtosecond (fs) pulsed lasers. A pair of ps-pulsed lights served for CARS (817 nm and 1064 nm) and SHG (817 nm) images; light from the fs-pulsed laser with the center wavelength of 720 nm was incident into the sample to obtain autofluorescence and targeted molecular TPEF images for high efficiency of fluorescence intensity without cross-talk. For multicolor-targeted TPEF imaging, we stained smooth-muscle cells and macrophages with fluorescent dyes (Alexa Fluor 350 and Alexa Fluor 594) for an immunofluorescence assay. Each depth-sectioned image consisted of $512{\times}512$ pixels with a field of view of $250{\times}250{\mu}m^2$, a lateral resolution of $0.4{\mu}m$, and an axial resolution of $1.3{\mu}m$. We obtained composite multicolor images with conventional label-free NLO images and targeted TPEF images in atherosclerotic-plaque samples. Multicolor 3-D imaging of atherosclerotic-plaque structural and functional composition will be helpful for understanding the pathogenesis of cardiovascular disease.

• Journal of the Korean Society for Precision Engineering
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• v.21 no.11
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• pp.75-82
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• 2004

A metrological atomic force microscope (M-AFM) was developed fur the length measurements of nanometer range, through the modification of a commercial AFM. To eliminate nonlinearity and crosstalk of the PZT tube scanner of the commercial AFM, a two-axis flexure hinge scanner employing built-in capacitive sensors is used for X-Y motion instead of PZT tube scanner. Then two-dimensional displacement of the scanner is measured using two-axis heterodyne laser interferometer to ensure the meter-traceability. Through the measurements of several specimens, we could verify the elimination of nonlinearity and crosstalk. The uncertainty of length measurements was estimated according to the Guide to the Expression of Uncertainty in Measurement. Among several sources of uncertainty, the primary one is the drift of laser interferometer output, which occurs mainly from the variation of refractive index of air and the thermal stability. The Abbe error, which is proportional to the measured length, is another primary uncertainty source coming from the parasitic motion of the scanner. The expanded uncertainty (k =2) of length measurements using the M-AFM is √(4.26)$^2$+(2.84${\times}$10$^{-4}$ ${\times}$L)$^2$(nm), where f is the measured length in nm. We also measured the pitch of one-dimensional grating and compared the results with those obtained by optical diffractometry. The relative difference between these results is less than 0.01 %.

• Korean Journal of Optics and Photonics
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• v.28 no.1
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• pp.9-15
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• 2017

We precisely measured and analyzed the dynamics of peptide-antibody interactions, using an ellipsometric biosensor based on a silicon substrate. To reduce the signal error due to the imperfect flatness of the substrate for extremely low concentrations of peptide, we fabricated the biosensor with a silicon substrate coated with Dextran SAM, instead of a glass prism coated with a thin metallic thin film. At an injection speed of $100{\mu}l/min$ of buffer liquid, we detected the dynamics of antibody-Dextran SAM or peptide-antibody fixed on biosensor, respectively. We detected the dynamics of antibody-Dextran SAM interactions down to a low concentration of 5 ng per liter, and we precisely measured the dynamics of association and dissociation of peptide and antibody down to 100 nM of peptide. We obtained the rate constants for association and dissociation from fitting the data by using deduced dynamical equation. As a result, we obtained an equilibrium constant for dissociation of 97 nM of peptide-antibody complex, which belongs to Class I.