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Imaging Biological Samples with Optical Microscopy01:18

Imaging Biological Samples with Optical Microscopy

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Optical microscopy uses optic principles to provide detailed images of samples. Antonie van Leeuwenhoek designed the first compound optical microscope in the 17th century to visualize blood cells, bacteria, and yeast cells. In 1830, Joseph Jackson Lister created an essentially modern light microscope. The 20th century saw the development of microscopes with enhanced magnification and resolution.
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Phase-Contrast Microscopes
In-phase-contrast microscopes, interference between light directly passing through a cell and light refracted by cellular components is used to create high-contrast, high-resolution images without staining. It is the oldest and simplest type of microscope that creates an image by altering the wavelengths of light rays passing through the specimen. Altered wavelength paths are created using an annular stop in the condenser. The annular stop produces a hollow cone of...
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Confocal microscopy is an advanced microscopic technique. The prime advantage of the confocal microscope over other microscopy techniques is its ability to block the out-of-focus light from the illuminated samples using pinholes. It is widely used with fluorescence optics to obtain high-resolution, sharp contrast images. Unlike optical microscopes, confocal microscopes use a focused beam of light laser to scan the entire sample surface at different z-planes. These microscopes are, therefore,...
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Total Internal Reflection Fluorescence Microscopy01:05

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Total internal reflection fluorescence microscopy or TIRF is an advanced microscopic technique used to visualize fluorophores in samples close to a solid surface with a higher refractive index, such as a glass coverslip. TIRF only allows fluorophores in proximity to the solid surface to be excited. When light from a medium with a lower refractive index (such as air) hits the glass coverslip at a critical angle, the light undergoes total internal reflection stead of passing through the glass.
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Super-resolution fluorescence microscopy (SRFM) provides a better resolution than conventional fluorescence microscopy by reducing the point spread function (PSF). PSF is the light intensity distribution from a point that causes it to appear blurred. Due to PSF, each fluorescing point appears bigger than its actual size, and it is the PSF interference of nearby fluorophores that causes the blurred image. Various approaches to achieving higher resolution through SRFM have recently been...
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マルチコンポーネント,リアルタイムイメージングのためのフローロゲンサイクロプロペノン

Tyler K Heiss1, Robert S Dorn1, Andrew J Ferreira1

  • 1Department of Chemistry, University of California, Irvine, California 92697, United States.

Journal of the American Chemical Society
|April 20, 2022
PubMed
まとめ
この要約は機械生成です。

サイクロプロペノンとフォスフィンを用いた新しいフローロゲンバイオオートホーゴン反応は,生細胞における生物分子のリアルタイムの可視化を可能にします. 細胞画像と生物学的標的の追跡に最適です 細胞画像と生物学的標的の追跡に最適です

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科学分野:

  • 化学生物学
  • バイオオートゴーナル・ケミストリー
  • 分子イメージング

背景:

  • フロオロゲン生物対称反応は,リアルタイムで生物分子の視覚化に不可欠です.
  • 既存の方法は,交叉反応性や信号強化が低いため,生細胞との互換性が欠けていることが多い.
  • 細胞イメージングには,高信号対ノイズ比を持つ新しい化学反応が必要である.

研究 の 目的:

  • 活体細胞のイメージングのための新しいフローロゲンバイオオートホーガン反応を開発する.
  • 信号の活性度が高く 交互反応が少ない
  • マルチカラーでリアルタイムの 画像処理を可能にします

主な方法:

  • バイオオルトゴン反応のためのサイクロプロペノンレポーターとフォスフィンパートナーの開発.
  • 地域選択的活性化とサイクル化を含む反応機構の調査.
  • 実験室内および生体細胞内での探査性能の評価
  • 多成分イメージングのための他のフッ素反応との互換性の評価.

主要な成果:

  • キュマリン製品を形成するサイクロプロペノンとフォスフィンの間の新しいフッ素反応が確立された.
  • 反応は1600倍以上のシグナルオンを示し, 報告された最高値の1つです.
  • 開発されたバイオオートホーゴナルモチーフは,試験管内および細胞環境で成功裏に検証されました.
  • この化学反応は他のフッ素反応と相容れていて,同時に画像を撮ることが可能になった.

結論:

  • サイクロプロペノン-フォスフィン反応は,バイオオートホゴン化学の非常に効率的で敏感な方法を提供します.
  • この反応は,ネイティブの細胞環境でリアルタイムで生物分子を追跡する能力を大幅に向上させます.
  • 開発された化学は,生細胞画像と多成分分析の範囲を拡大するための貴重なツールです.