Media 4: Multi-focus excitation coherent anti-Stokes Raman scattering (CARS) microscopy and its applications for real-time imaging

Originally published in Optics Express on 08 June 2009 (oe-17-12-9526

Schematic of cavity-reflection-enhanced absorption microscopy (CREAM).

<p>(A) Optical system of CREAM. SCLaser, supercontinuum laser; RC, reflective collimator; SPF, short pass filter; L1 to L5, spherical achromatic lenses; PH, pinhole; CM, cavity mirror; PZT, piezo xy-stage; SU, spectrum unit; CCD, charge coupled-device. Dotted box area indicates optical cavity. We set optical cavity length L = 120 mm for concentric-type optical cavity. (B) Focused spot images of before (left) and after (right) adjustment in the center of cavity measured by CCD camera. Scale bar represents 50 μm.</p

Media 1: Multi-focus excitation coherent anti-Stokes Raman scattering (CARS) microscopy and its applications for real-time imaging

Originally published in Optics Express on 08 June 2009 (oe-17-12-9526

Spectral Fingerprinting of Individual Cells Visualized by Cavity-Reflection-Enhanced Light-Absorption Microscopy

<div><p>The absorption spectrum of light is known to be a “molecular fingerprint” that enables analysis of the molecular type and its amount. It would be useful to measure the absorption spectrum in single cell in order to investigate the cellular status. However, cells are too thin for their absorption spectrum to be measured. In this study, we developed an optical-cavity-enhanced absorption spectroscopic microscopy method for two-dimensional absorption imaging. The light absorption is enhanced by an optical cavity system, which allows the detection of the absorption spectrum with samples having an optical path length as small as 10 μm, at a subcellular spatial resolution. Principal component analysis of various types of cultured mammalian cells indicates absorption-based cellular diversity. Interestingly, this diversity is observed among not only different species but also identical cell types. Furthermore, this microscopy technique allows us to observe frozen sections of tissue samples without any staining and is capable of label-free biopsy. Thus, our microscopy method opens the door for imaging the absorption spectra of biological samples and thereby detecting the individuality of cells.</p></div

Media 3: Multi-focus excitation coherent anti-Stokes Raman scattering (CARS) microscopy and its applications for real-time imaging

Originally published in Optics Express on 08 June 2009 (oe-17-12-9526

Media 2: Multi-focus excitation coherent anti-Stokes Raman scattering (CARS) microscopy and its applications for real-time imaging

Originally published in Optics Express on 08 June 2009 (oe-17-12-9526

Observation of human rectal tumor model and mouse normal rectal tissues.

<p>(A) Hematoxylin-Eosin stained mouse normal rectal tissues (left) and human rectal tumor model (right). Scale bar represents 0.5 mm. (B) Cavity-reflection enhanced absorption images of mouse normal rectal tissues (left) and human rectal tumor model (right). Scale bar represents 100 μm. Pseudo-colored images were constructed by merging the 3 colors as follows: blue for 450 to 500 nm, green for 500 to 550 nm, and red for 550 to 600 nm of averaged images. (C) Cavity-reflection-enhanced absorption spectrum of mouse normal rectal tissues (black) and human rectal tumor model (red).</p

Light-absorption spectrum of Venus yellow fluorescent protein by CREAM.

<p>Various concentrations of Venus fluorescent protein with different sample thicknesses of (A) 100-μm and (B) 10-μm.</p

Absorption images and spectra of various cell types.

<p>(A) Cavity-reflection-enhanced absorption images for cell types as indicated. Pseudo-colored images were constructed by merging the 3 colors as follows: blue for 450 to 500 nm, green for 500 to 550 nm, and red for 550 to 600 nm of averaged images. Scale bars represent 100 μm. (B) Averaged spectrum of each cell type (n = 10 for each cell type). Colors correspond to the different cell types in Fig 3B. (C) Scatter plot of PCA scores by PC1 and PC2 components. Error bars represent standard deviation.</p

Nanometer-precision surface metrology of millimeter-size stepped objects using full-cascade-linked synthetic-wavelength digital holography using a line-by-line full-mode-extracted optical frequency comb

Digital holography (DH) is a powerful tool for surface profilometry of objects with sub-wavelength precision. In this article, we demonstrate full-cascade-linked synthetic-wavelength DH (FCL-SW-DH) for nanometer-precision surface metrology of millimeter-size stepped objects. 300 modes of optical frequency comb (OFC) with different wavelengths are sequentially extracted at a step of mode spacing from a 10GHz-spacing, 3.72THz-spanning electro-optic modulator OFC (EOM-OFC). The resulting 299 synthetic wavelengths and a single optical wavelength are used to generate a fine-step wide-range cascade link covering within a wavelength range of 1.54 um to 29.7 mm. We determine the 0.1000mm-stepped surface with axial uncertainty of 6.1 nm within the maximum axial range of 14.85 mm