Name: 単一分子局在顕微鏡を用いた細胞核中の絶対DNA密度のマッピング (動画) | JoVE Journal
Uploaded: 2025-11-11T14:00:00.000+00:00
Duration: 10 min 57 s
Description: 学ぶ単一分子局在顕微鏡とVoronoi分割を用いた細胞核内絶対DNA密度測定法。染色質構造の空間的制約を解析し、細胞周期段階とDNA含量の関係を明らかにする。

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</ol>

<table><tbody><tr><td><強さ>細胞培養<強さ></td><td></td><td></td><td></td></tr><tr><td>&micro;-ディッシュ 35 mm、ハイグリッド500ガラス底</td><td>イビディ</td><td>81168</td><td></td></tr><tr><td>C3H 10T1/2</td><td>IMB(ニールズ・ラボ)</td><td></td><td></td></tr><tr><td>DMEM</td><td>サーモフィッシャー</td><td>12320032</td><td></td></tr><tr><td>dPBS</td><td>サーモフィッシャー</td><td>14190144</td><td></td></tr><tr><td>FBS</td><td>ライフ・テクノロジーズ</td><td>16000-044</td><td></td></tr><tr><td>ヘラ</td><td>顕微鏡コア施設(IMB)</td><td></td><td></td></tr><tr><td>HFB</td><td>IMB(Kubben Lab)</td><td></td><td></td></tr><tr><td>L-グルタミン</td><td>シグマ・オルドリッチ</td><td>G7513</td><td></td></tr><tr><td>HFBに対する非必須アミノ酸とビタミン</td><td></td><td></td><td></td></tr><tr><td>ピルビン酸ナトリウム</td><td>S8636</td><td></td><td></td></tr><tr><td><ストロング>サンプル準備</ストロング></td><td></td><td></td><td></td></tr><tr><td>カタラーゼ</td><td>メルク</td><td>2593710</td><td></td></tr><tr><td>グルコース</td><td>サーモフィッシャー</td><td>241922500</td><td></td></tr><tr><td>グルコース酸化酵素</td><td>メルク</td><td>49180</td><td></td></tr><tr><td>パラホルムアルデヒド</td><td>シグマ・オルドリッチ</td><td>158127</td><td></td></tr><tr><td>RNaseカクテル</td><td>サーモフィッシャー</td><td>AM2286</td><td></td></tr><tr><td>SYTOX オレンジ</td><td>サーモフィッシャー</td><td>S11368</td><td></td></tr><tr><td>テトラスペック蛍光微小球サンプラーキット</td><td>サーモフィッシャー</td><td>T7284</td><td></td></tr><tr><td>トライトン X-100</td><td>サーモフィッシャー</td><td>327372500</td><td></td></tr><tr><td>Software</td><td></td><td></td><td></td></tr><tr><td>バイオフォーマット</td><td>OpenMicroscopy.org</td><td></td><td>オープンソースソフトウェア https://www.openmicroscopy.org/bio-formats/</td></tr><tr><td>CellProfiler v4.2.1</td><td>CellProfiler.org</td><td></td><td>オープンソースソフトウェア https://cellprofiler.org</td></tr><tr><td>フィジー</td><td>nih.gov</td><td></td><td>オープンソースソフトウェア https://imagej.net/software/fiji/?Downloads</td></tr><tr><td>陸</td><td>nih.gov</td><td></td><td>オープンソースソフトウェア https://github.com/Jan-NM/LAND</td></tr><tr><td>マットラボ 2021</td><td>数学作品</td><td></td><td>商用ソフトウェア - 「イメージ処理ツールボックス」が必要</td></tr><tr><td>R.4...0.3</td><td>r-project.org</td><td></td><td>オープンソースソフトウェア https://www.r-project.org</td></tr><tr><td>サンダーストーム v1.3</td><td></td><td></td><td>オープンソースソフトウェア https://zitmen.github.io/thunderstorm/</td></tr><tr><td><ストロング>顕微鏡:</ストロング></td><td></td><td></td><td></td></tr><tr><td>AF 7000</td><td>ライカ</td><td></td><td></td></tr><tr><td>ライカGSD</td><td>ライカ</td><td></td><td></td></tr><tr><td>SMLM顕微鏡</td><td>クレマー研究所</td><td></td><td>S-Y博士が特注製作した。陳</td></tr></tbody></table>

単一分子局在顕微鏡を用いた細胞核中の絶対dna密度のマッピング

Mapping Absolute DNA Density in Cell Nuclei using Single-molecule Localization Microscopy

単一分子局在顕微鏡を用いた細胞核中の絶対DNA密度のマッピング

Within the cell nucleus, silent genes are generally located in chromatin areas of high density called heterochromatin, whereas active genes can be mostly ...

mapping-absolute-dna-density-cell-nuclei-using-single-molecule

Research

JoVE Journal

Biology

生物学

単一分子顕微鏡は、核輸送への新たな洞察を提供approch。...

核輸送の1分子イメージング

私たちは、直接標識蛍光プローブを用いた3次元のDNA...

直接標識プローブを用いた堅牢な3DのDNA FISH

単一分子追跡と組み合わさ光活性化局在化顕微鏡（PALM）は、生大腸菌細胞におけるタンパク質-DNA相互作用の直接観察及び定量化を可能にする。...

光活性化単一分子追跡を使用してライブ細菌細胞におけるタンパク質-DNA相互作用を可視化する

蛍光標識された細胞の多重化された画像ベースの分析を可能に柔軟インフォマティクスワークフローがある発表。ワークフローは、核と細胞質のマーカーを定量化し、これらの区画の間のマーカー座を計算します。手順は、96ウェルフォーマットで、間接免疫蛍光法によってマーカーを検出するためのsiRNAで信頼性の高い方法論を用いた細胞の摂動のために提供される。...

高コンテンツ、オープンソースソフトウェアでサポートされているユニバーサル顕微鏡データからの蛍光マーカー、個々の細胞定量のワークフロー

新たに発見された 5-メチルシトシン (oxi mCs), 5-ヒドロキシメチルシトシン (5hmC), 5-formylcytosine (5 fC) の酸化フォームと 5-carboxylcytosine (5caC) は、独自の機能的役割の異なる DNA 変更を表す場合があります。ここの可視化から oxi mCs...

DNA 変更染色: 共焦点画像の数値解析

3D単分子局在顕微鏡は、生きた細菌細胞における蛍光標識タンパク質の空間位置および運動軌道をプローブするために利用される。本明細書に記載される実験およびデータ分析プロトコルは、プールされた単一分子軌道に基づいてサイトソリックタンパク質の一般的な拡散挙動を決定する。...

単一分子トラッキング顕微鏡 - 細胞細胞分子の拡散状態を決定するツール

ヒト初等細胞の核アーキテクチャを研究する3D多色DNA FISHツール

このプロトコルは、細胞膜タンパク質のナノスケール空間的組織を解消するための単一抗体標識(SAL)を記述しています。抗体とエピトープの累積相互作用を単一分子レベルで活用することで、膜SAL(mSAL)は局所的なエピトープ分布をマッピングしつつ、同時にネイティブ細胞環境における抗体結合挙動を捉えます。...

膜タンパク質の単一分子局在顕微鏡法による単一抗体標識法

私たちは、ユークロマチン、ヘテロクロマチン、RNAP...

高密度核環境下のクロマチン調査のための多ラベル単分子局在顕微鏡プロトコル

このプロトコルは、DNA内在蛍光スペクトルのサンプル準備、データ収集、解析に関する詳細な操作を提供します。...

内在コントラストを用いたDNA分子の分光超分解能イメージング

このプロトコルは、最適化された分離、固定、標識を用いてクロマチンおよびRNAポリメラーゼIIの信頼性の高いナノスケール可視化を実現し、単離シロイナズナの核の単分子局在顕微鏡イメージングの効率的なワークフローを提供します。この手法は、他のクロマチン関連タンパク質や高分解能イメージングのためのヒストン修飾にも容易に適応可能です。...

シロイナナズナ核における単一分子局在顕微鏡の効率的なプロトコル

学ぶ単一分子局在顕微鏡とVoronoi分割を用いた細胞核内絶対DNA密度測定法。染色質構造の空間的制約を解析し、細胞周期段階とDNA含量の関係を明らかにする。

単一分子局在顕微鏡を用いた細胞核中の絶対DNA密度のマッピング

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