高分子　Vol.64　No.9　2015年9月

高分子　Vol.64　No.9

特集　超解像蛍光顕微鏡と高分子

展望　COVER STORY: Highlight Reviews

PALM用の蛍光タンパク質と生物学での応用
Photoswitchable Fluorescent Proteins for PALM: Molecular Basis and Tips for Labeling

水野　秀昭
Hideaki MIZUNO

＜要旨＞蛍光タンパク質は生細胞内で非侵襲的に分子を標識するツールである。光スイッチング蛍光タンパク質の発見により、その応用範囲は分子トラッキングや顕微鏡上でのハイライティングに広がり、また光スイッチング蛍光タンパク質はPALMの開発にも貢献した。本稿ではPALMに用いられる光スイッチング蛍光タンパク質について概説する。
Keywords: Photoswitchable Fluroescent Protein / Super-Resolution Microscopy / PALM

ページトップへ▲

非点収差を用いた高分子の三次元ダイナミックスの評価

Super-Resolution Localization Microscopy Reveals Nanoscale Dynamics of Polymers and Soft Matter

平田　修造・伊都　将司・Martin Vacha・宮坂　博
Shuzo HIRATA, Syoji ITO, Martin VACHA, Hiroshi MIYASAKA

＜要旨＞材料薄膜は厚さがナノメートルスケールになると表面や基板界面の影響を強く受け、拡散や回転運動などが膜厚方向とほかの方向とで大きく異なる。本稿では非点収差顕微鏡を用いて膜厚方向のダイナミックスを評価する手法を解説する。さらに高分子薄膜や液晶薄膜を例にとり、膜厚方向のダイナミックスを評価した最近の研究を紹介する。
Keywords: Super-Resolution Microscopy / Single Molecule / Polymer Film / Relaxation Dymanics / Diffusion / Liquid Crystal

ページトップへ▲

単一分子局在化顕微鏡の高分子鎖のコンホメーション解析への応用
Conformational Analysis of Polymer Chain by Single Molecule Localization Microscopy

青木　裕之
Hiroyuki AOKI

＜要旨＞単一分子局在化顕微鏡に開発によってナノメートルスケールの空間分解能での蛍光イメージングが実現した。これによりバルク材料内部の構造とダイナミックスを三次元的に単一分子レベルで評価することができるようになった。本稿では単一分子局在化顕微鏡の原理を概説するとともに、高分子鎖のコンホメーション解析への応用について述べる。
Keywords: Single Molecule Localization Microscopy / Super-Resolution / Single Polymer Chain / Conformation / Deformation / Fluorescence

ページトップへ▲

トピックス　COVER STORY: Topics and Products

STED顕微鏡の原理とその応用例
Principle and Application of STED Microscopy

堀田　純一
Jun-ichi HOTTA

＜要旨＞ Stimulated emission depletion (STED) microscopy is one of the super-resolution fluorescence microscopy techniques, which realize better resolution than the diffraction limit of light. In order to squeeze a fluorescent spot, STED microscopy utilize fluorescence depletion by stimulated emission process for switching off the fluorescence molecules except for the center of the donut STED beam. In this paper, principle and application of STED microscopy are discussed. The important role of patterned polymer thin films as spatical phase modulators is explained. Generation of two- and three-dimensional donut beams using polymer thin films are described. As examples of STED microscopy in the field of polymers, single molecule imaging of dendrimers, visualization of nanoscale block copolymer morphology, and STED lithography are discussed.
Keywords: STED Microscopy / Super-Resolution / Dendrimer / Block Copolymer / STED Lithography

ページトップへ▲

分子吸着を用いた超解像イメージング
Super-Resolution Fluorescence Imaging Based on Molecular Adsorption

羽渕　聡史
Satoshi HABUCHI

＜要旨＞ While super-resolution fluorescence imaging has been revolutionizing the way we study nanoscale structures, applications of the imaging technique is still limited, partly due to limited fluorophores and labeling methods available for the imaging. Here, I introduce a localization microscopy technique (PALM/STORM) based on an adsorption of fluorophores to a target structure. This method does not require a covalent labeling of a target structure with a special fluorophore whose fluorescent state can be switched, which is not a trivial issue. In this approach, the fluorophores adsorb selectively to the target structure. Fluorescent states of the fluorophore are then spatiotemporally controlled by adsorption/desorption kinetics of the fluorophore. Based on the adsorption/desorption-induced fluorescence switching, a super-resolution image is reconstructed, similar to PALM/STORM imaging. I show several examples of the super-resolution fluorescence imaging based on this approach, including micro phase separation of phospholipids in a lipid bilayer, DNA nanostructures, and polymer nanostructures.
Keywords: Super-Resolution / Fluorescence Imaging / STORM / Single-Molecule / Adsorption / Nano-Structure / Thermal Nanoimprinting

ページトップへ▲

構造化照明法超解像顕微鏡の原理と応用
The Principle and the Applications of Structured Illumination Super Resolution Microscopy

及川　義朗
Yoshiro OIKAWA

＜要旨＞ Optical microscopy has the theoretical resolution limit of 200 nm which is defined as d=λ/2NA. Recently some ideas of breaking this limit are employed in the commercially available product, known as “super resolution microscopy”. “Structured Illumination Microscopy (SIM)” is one of those technologies that provides approx. 100 nm resolution. In the SIM method, a stripe illumination pattern is applied to capture the diffraction light. Several images with such illumination pattern are captured and they are converted into Fourier space. Then high frequency components are separated and relocated. The image frequency range has become twice bigger. Then they are converted back into the real space, and a super resolution image with some 100 nm resolution is generated. The Electron microscope is useful to observe a very small sample, but not applicable to a living sample. SIM is used in the fields of neuro-science, immunology research or regenerative medicine research. Super resolution microscopy is expanding its application field not only for bioscience, but material science and other fields.
Keywords: Microscope / Super Resolution / SIM / Structured Illumination

ページトップへ▲

赤外超解像顕微鏡法
IR Super-resolution Microscopy

酒井　誠・藤井　正明
Makoto SAKAI, Masaaki FUJII

＜要旨＞ For many years, spatial resolution was the most critical problem in IR microscopy. This is because the spatial resolution of a conventional IR microscope is restricted by the diffraction limit, which is almost the same as the wavelength of IR light, ranging from 2.5 to 25 μm. In the recent years, we have developed two novel types of far-field IR super-resolution microscopes using 2-color laser spectroscopies, transient fluorescence detected IR spectroscopy and vibrational sum-frequency generation (VSFG) spectroscopy. Applying these methods, in which both transient fluorescence and VSFG signal have a wavelength in the visible region, the image is observed at the resolution of visible light, which is about 10 times smaller than that of IR light, that is, IR super-resolution. By using these techniques, we can map the specific IR absorption with a sub-micrometer spatial resolution, visualization of the molecular structure includung molecular orientations in a non-uniform environment becomes a possibility.
Keywords: IR Super-Resolution / Microscope / 2-Color Laser Spectroscopy / IR Imaging / IR Spectroscopy / Molecular Structure / Molecular Orientation

ページトップへ▲

グローイングポリマー　Polymer Science and I: A Personal Account

受けポジ
Passive but Positive

中嶋　琢也
Takuya NAKASHIMA

＜要旨＞ I had great opportunities to be involved in the startup of new laboratories by chance. These experiences made me open to new research fields and a witty player. I’m still struggling with science to achieve a great discovery.

ページトップへ▲

高分子科学最近の進歩　Front-Line Polymer Science

高分子ミセルを鋳型とする無機中空ナノ粒子の合成
Synthesis of Inorganic Hollow Nanoparticles Templated by Polymeric Micelles

中島　謙一
Kenichi NAKASHIMA

＜要旨＞ Inorganic hollow nanoparticles have many applications from use as heat insulation and lightweight materials to drug carriers for medicinal purposes. Two synthesis techniques have been used so far; one relies on a template and the other is template-free. Template-free processes have a drawback that it is difficult to control particle size. The templates are further classified into two; one is hard and the other is soft. Hard template techniques also suffer from low product yield and shell weakness. Thus, researchers have turned to soft templates, especially polymeric micelles. In this review, we introduce recent progress in the synthesis of inorganic hollow nanoparticles focusing on a strategy of polymeric micelle templates. We demonstrate advantages of ABC triblock copolymer micelles with core-shell-corona architecture. By tailoring experimental parameters, we can readily obtain a variety of hollow nanoparticles including silica, metal-oxides, metal-phosphates, etc. Finally, we highlight the potential use of these hollow particles in drug delivery and bio-imaging.
Keywords: Inorganic Hollow Nanopartilce / Template Syntehsis / Polymeric Micelle

ページトップへ▲