Name: micropipette aspiration of substrate-attached cells to estimate cell stiffness
Uploaded: 2012-09-27T12:00:00
Description: Watch this Scientific Journal Video about Micropipette Aspiration of Substrate-attached Cells to Estimate Cell Stiffness at JoVE.com

1。ガラスマイクロピペットを引っ張る 機器：マイクロピペットプラー、マイクロフォージ。 ガラス：Boroscillicateガラスキャピラリー（〜1.5ミリメートル外径は、〜1.4ミリメートル内径）。 <ol><li> マイクロピペットは、電気生理学的記録のためにガラス微小電極を準備するために使用されているのと同じ基本?...

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Microaspirationは、細胞膜に負圧を適用し、明確に定義された圧力に応答して膜の変形能を測定することにより細胞の剛性/変形を推定するためのシンプルで再現性の高い方法を提供します。これは、最初の細胞分裂21その後赤血球1の機械的特性を見るためのメカニズムへの洞察を提供するために、ウニの卵の弾性的性質を特徴づけるためにミッチソンとスワン（1954）によって開...

<table border="1"><tbody><tr><td> 試薬の名称 </td><td> 会社 </td><td> カタログ/モデル番号 </td><td> 注釈 </td></tr><tr><td>サッターピペットプラー</td><td>サターインスツルメンツ</td><td> P-97 </td><td></td></tr><tr><td>マイクロフォージ</td><td>ナリシゲ</td><td> MF-830 </td><td></td></tr><tr><td>倒立蛍光顕微鏡</td><td>ツァイス</td><td> AXIOVERT 200M </td><td>顕微鏡は、好ましく3D/deconvolution機能を装備する必要があります。 </td></tr><tr><td>ビデオカメラ</td><td>ツァイス</td><td> AxioCam MRM </td><td></td></tr><tr><td>画像取得sotware </td><td>ツァイス</td><td> AxioVisionの</td><td></td></tr><tr><td>空気圧トランスデューサ</td><td> BioTekの</td><td> DPM-1B </td><td> DPM1B空変換器のテスターは現在FLUKEことによって見つけることができます。 </td></tr><tr><td>ピペットグラス</tD&gt; <td>リッチランド</td><td>カスタマイズされたガラス</td><td>ピペットは1.2内径と外径1.6を使用してカスタマイズされた。 </td></tr><tr><td> DiIを染料</td><td>インビトロジェン</td><td> D282 </td><td> DMSOによく溶ける</td></tr></tbody></table>

micropipette aspiration of substrate-attached cells to estimate cell stiffness

研究の数が増え、個々の細胞の生体力学的特性は、細胞の増殖、分化、移動および細胞間相互作用を含む複数の細胞機能において重要な役割を果たしていることを示している。細胞のバイオメカニクスの2つの重要なパラメータは、細胞変形や剛性と力を収縮し、生成する細胞の能力である。ここでは、細胞表面にガラスマイクロピペット、マイクロピペット吸引またはMicroaspirationと呼ばれる技術によって適用された負圧に応じて、膜変形の度合いを測定することにより、セルの剛性を推定するために迅速かつ簡単な方法を説明します。 Microaspirationその後、空気圧トランスデューサに接続されており、近くに持って来られる、非常に小さなチップ（細胞または組織サンプルの大きさに応じて2から50μmの直径）でマイクロピペットを作成するためにガラスキャピラリーを引っ張ることによって行われます顕微鏡下で細胞の付近。時ピペットの先端が細胞に触れると、負圧工程は、細胞膜上の明確に定義された圧力を発生させる空気圧トランスデューサによりピペットに適用されます。圧力に応じて、メンブレンをピペット内に吸引され、ピペットにプログレッシブ膜変形または &quot;膜投影&quot;は、時間の関数として測定される。この実験的なアプローチの基本的な原理は、定義された機械的な力に応じて、膜変形の程度は膜剛性の関数であるということです。硬めの膜であり、膜変形と短い定常吸引length.The技術の遅い速度は、単離された両方のサスペンションと、基板に接続された、大規模な細胞小器官内の細胞、およびリポソーム上で実行することができます。 分析は、異なる細胞集団や実験条件の与えられた圧力の下で達成最大の膜の変形を比較することによって行われる。 &quot;剛性係数は、&quot; ESです印加圧力の関数としての膜変形の吸引された長さをプロットすることによりtimated。さらに、データはさらに材料の剛性を特徴づける細胞のヤング率（E）は、最も一般的なパラメータを推定するために分析することができる。真核細胞の細胞膜は膜脂質二重層は、サブ膜細胞骨格によって、それが膜の機械的足場を構成し、変形を支配する細胞骨格であるunderliedされる二成分系とみなすことができることに注意することが重要です携帯封筒の。このアプローチは、したがって、サブ膜細胞骨格の力学的特性をプロービングすることができます。

Watch this Scientific Journal Video about Micropipette Aspiration of Substrate-attached Cells to Estimate Cell Stiffness at JoVE.com

Micropipette Aspiration of Substrate-attached Cells to Estimate Cell Stiffness

ここでは、セルの剛性を測定するために迅速かつ簡単な方法を説明します。このアプローチの一般的な原則は、細胞表面にマイクロピペットを介して適用、明確に定義された負圧に応答して膜の変形を測定することです。この方法は、基板に接続された細胞の生体力学的特性を研究するための強力なツールを提供します。

セルの剛性を推定するために基板に接続された細胞のマイクロピペット吸引

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Transplantation of Induced Pluripotent Stem Cell-derived Mesoangioblast-like Myogenic Progenitors in Mouse Models of Muscle Regeneration

筋肉再生のマウスモデルにおける誘導多能性幹細胞由来メソアンジオ芽細胞様筋原性前駆細胞の移植

Patient-derived iPSCs could be an invaluable source of cells for future autologous cell therapy protocols. iPSC-derived myogenic stem/progenitor cells ...

Ordering Single Cells and Single Embryos in 3D Confinement: A New Device for High Content Screening

シングルセルおよび3D閉じ込めにおける単一胚を注文：ハイコンテントスクリーニングのための新しいデバイスを

Biological cells are usually observed on flat (2D) surfaces. This condition is not physiological, and phenotypes and shapes are highly variable. Screening ...

Simple Polyacrylamide-based Multiwell Stiffness Assay for the Study of Stiffness-dependent Cell Responses

剛性依存性細胞応答の研究のための簡単​​なポリアクリルアミド系マルチウェル剛性アッセイ

Currently, most of the in vitro cell research is performed on rigid tissue culture polystyrene (~1 GPa), while most cells in the body are attached to a ...

A Method to Estimate Cadaveric Femur Cortical Strains During Fracture Testing Using Digital Image Correlation

破壊を使用してデジタル画像相関法を試験中に死体大腿骨皮質系統を推定する方法

This protocol describes the method using digital image correlation to estimate cortical strain from high speed video images of the cadaveric femoral ...

Human Pluripotent Stem Cell Culture on Polyvinyl Alcohol-Co-Itaconic Acid Hydrogels with Varying Stiffness Under Xeno-Free Conditions

Xeno 無料条件下におけるさまざまな剛性とポリビニル アルコール Co イタコン酸ゲルのひと多能性幹細胞文化

The effect of physical cues, such as the stiffness of biomaterials on the proliferation and differentiation of stem cells, has been investigated by ...

Stiffness Measurement of Soft Silicone Substrates for Mechanobiology Studies Using a Widefield Fluorescence Microscope

広視野蛍光顕微鏡を用いたメカノバイオロジー研究の柔らかいシリコーン基板の剛性測定

Soft tissues in the human body typically have stiffness in the kilopascal (kPa) range. Accordingly, silicone and hydrogel flexible substrates have been ...

Single-Cell Optical Action Potential Measurement in Human Induced Pluripotent Stem Cell-Derived Cardiomyocytes

ヒト人工多能性幹細胞由来心筋細胞における単細胞光学作用電位測定

Conventional intracellular microelectrode techniques to quantify cardiomyocyte electrophysiology are extremely complex, labor intensive, and typically ...