Name: hemi-laryngeal setup for studying vocal fold vibration in three dimensions
Uploaded: 2017-11-25T13:00:00.000+00:00
Duration: 10 min 13 s
Description: Watch this Scientific Journal Video about Hemi-laryngeal Setup for Studying Vocal Fold Vibration in Three Dimensions at JoVE.com

この作品が支持されたオーストリアのアカデミー科学 (CTH) のアパートの付与、チェコ共和国の技術振興プロジェクトなし。TA04010877 (CTH、VH と国債)、およびチェコ科学財団 (GACR) プロジェクト (国債) に GA16 01246S のないです。義歯定着性のクリームを使用する彼の提案のためのテカムセ フィッチ w. と Ing に感謝しますP. リシュカ摘出鹿 larynges を取得するのに彼の助けをチェコ軍のフォレスト サービスから。

本稿で分析した動物の標本は、パラツキー大学オロモウツ、チェコ共和国での標準倫理的な要件に従って扱われました。彼らは赤いシカの通常の狩猟シーズン中にチェコ軍森林サービスによって捜された林で乱暴生活から生じる。
1. 片側喉頭標本の準備
注: 唯一の適切に準備された標本使わなければなりません、4に示すように。喉頭の凍結迅速な切除し-80 ° C で保存した直後に25組織分解の潜在性と力学的特性の変化を最小限に抑えるし、でいつでも実験を実行することができます。
<ol><li>
喉頭を解凍
	<ol>
<li>2 オートクレーブ バッグや防水シールと他のビニール袋に冷凍の喉頭を挿入します。袋を密封して喉頭を完全に解凍するまで 30 の ° C に加熱水のお風呂に入れます。必要な期間の範囲は、数時間から喉頭サイズおよび凍結温度によってより多くの日。</li>
	</ol>
</li>
	<li>
喉頭をクリーニング
	<ol>
<li>バッグから削除し、徹底的にそれをきれいに生理食塩液を喉頭を解凍後 (0.9 %nacl)。</li>
		<li>慎重に必要に応じて、余分な組織を削除 (すなわち外部の首の筋肉、舌骨など) なしメインの喉頭の構造の損傷や気管、空気供給管に喉頭をマウントするために十分な長さを短縮 (ca. 通常 4-5 cm)。</li>
		<li>潜在的実験には不向き、喉頭を作ることができる凍結の過程から発生するクラックや有機の変形、傷などの潜在的な生体組織異常のため喉頭組織を確認します。</li>
	</ol>
</li>
	<li>
甲状腺と輪状軟骨の露出
	<ol>
<li>正中カット片側喉頭の作成の準備で軟骨を公開するので、メスを用いた甲状腺と輪状軟骨周囲の外喉頭筋組織の部分を削除します。この準備段階は、図 1 aおよび1 bで描かれています。</li>
	</ol>
</li>
	<li>
正中甲状軟骨を切ってください。
	<ol>
<li>甲状軟骨の前部の初期垂直カットを作る。</li>
		<li>保存状態を維持する必要がある声帯に損傷を与えるために、削除される側に少しカットして慎重に置きます。可能であれば、切断用メスを使用します。軟骨が骨化すると場合、は、小さな鋸を使用します。</li>
	</ol>
</li>
	<li>
輪状軟骨を切断
	<ol>
<li>垂直方向にカットをリード (下方に) から披裂軟骨の間に放置して下甲状腺ノッチの約水平レベルに輪状軟骨。</li>
	</ol>
</li>
	<li>
喉頭に L 字切開を作成する 1 つの声帯除去
	<ol>
<li>輪状軟骨で、以前作った垂直カットの下の端から始まってを切り取って、下甲状腺切り込みに向かって新しいカットを導く水平を確認します。前方削除する起こっている喉頭の側面を隠します。</li>
		<li>甲状軟骨の内側の軟部組織を切って、甲状軟骨、声帯への損傷を避けるために、声帯の前方の添付ファイルの間にカットをリードしながら注意してください - 垂直を確認します。</li>
	</ol>
</li>
	<li>
甲状軟骨のカットの絞り込み
	<ol>
<li>以前検査した声帯の前方部分にできるだけ近いメス、のこぎり、または甲状軟骨で正確にストレート カットを適用し、として取得するために、ファイルを使用します。</li>
		<li>また後部の甲状軟骨の一部を削除、披裂の内転の芯を挿入するためのスペースを作成するために軟骨そしてこうしてボーカル折り (下記参照)。この準備段階は、図 1と1 Dで描かれています。 注: 研究の質問によって声帯全体における博覧会は上からの表示を有効にするのに必要かもしれない。このような場合は、(真) の声帯 (すなわち、心室または前庭倍、解剖学標本の指定に応じて) 上の構造を削除してください。いくつかの標本は声帯より上内側の軟喉頭部甲状軟骨との接続を失う可能性があり、声帯振動時に干渉、スプリアスの (大抵不規則な) 振動パターンを引き起こして可能性があります。このようなケースでは、その組織を慎重に除去は避けられないです。</li>
	</ol>
</li>
</ol><img alt="Figure 1" class="xfigimg" src="/files/ftp_upload/55303/55303fig1.jpg"> 図 1: ヘミ喉頭準備および取り付け。(A) と (B) クリーニング喉頭片、左声帯; 削除する前に、内側と後部のビュー(C) と (D) 準備されたヘミ喉頭 L 字切開 (左声帯除去)、内側と後部のビュー。<a href="//cloudflare.jove.com/files/ftp_upload/55303/55303fig1large.jpg" target="_blank">この図の拡大版を表示するのにはここをクリックしてください</a>。

2. 片側喉頭実験
<ol><li>
ヘミ喉頭セットアップ
	<ol>
<li>喉頭に温め、加湿空気を提供する空気供給管を使用します。</li>
		<li>構造 2 は垂直に削除された喉頭部分の代替として透明板を配置しました。</li>
		<li>使用熊手が喉頭の安定性が向上し、縦のガラス プレートに残りの声帯の内転によって適切な pre-phonatory 喉頭構成を作成するため4 (図 2 a参照)。 注: 理論的には、声帯可能性がありますもが内転し縫合糸と滑車レバー システム26の重みで。しかし、このようなアプローチは、これらの著者の最高の知識にまだ試みられて hemilarynx 準備のため。</li>
	</ol>
</li>
</ol><img alt="Figure 2" class="xfigimg" src="/files/ftp_upload/55303/55303fig2.jpg">
図 2: ヘミ喉頭セットアップします。(A) 支持構造: 空気供給管、L 型のガラス プレート アレンジ、内転の熊手。(B) 片側喉頭内転の突起を持つ準備をマウントします。(C)、ヘミ-喉頭-準備の (D) クローズ アップ表示側からと、上からそれぞれ。<a href="//cloudflare.jove.com/files/ftp_upload/55303/55303fig2large.jpg" target="_blank">この図の拡大版を表示するのにはここをクリックしてください</a>。
<ol start="2"><li>
ヘミ喉頭をマウント
	<ol>
<li>義歯固定剤クリームを空気供給管をカバーし、喉頭、気管の残りの部分をマウントします。定着性のクリームは接着剤として働き、従って気密のシールを作成する、潜在的なギャップを閉じます。</li>
		<li>プラスチック製のストラップやホースのクランプを締めと気管を固定します。</li>
		<li>また定着剤の定着性声帯や喉頭生体組織の内部ソフト クリームを拡散を回避しながら、クリームと甲状軟骨のカットのエッジをカバーします。</li>
		<li>透明の板を取り付けます。</li>
	</ol>
</li>
	<li>
甲状軟骨の突起を使用して声帯の内転の安定化
	<ol>
<li>付加体をプレートに声帯、甲状軟骨を安定させる突起を使用します。</li>
		<li>定着性のクリームが設定後は、声帯振動とヘミ喉頭とガラス板の可能なリークのチェックを確立するために空気の流れを適用します。</li>
		<li>多く定着性のクリームを追加することによって、最終的に発生するギャップをシールします。</li>
	</ol>
</li>
</ol>

著者が明らかに何もありません。

ヘミ喉頭準備を共有する「従来型」の (完全な) 摘出喉頭セットアップの利点: このような実験的アプローチ、物理的、生理的な境界条件とパラメーター (声門圧力や声帯伸長) などはすることができますかなりよく制御されます。Hemilarynx の動作は相同性と完璧な完全喉頭の横方向の対称性、例外とその大きさのいくつかのパラメーターの (e.g、風量、音圧) 約 50% 削減はまだまだ内にあ?...

声は、空気パルス シーケンスに肺によって提供される安定した気流を変換する喉頭組織 (主に声帯ヒダ) を振動によって通常作成されます。音圧波形 (つまり、プライマリの音)、それらをフィルタ リングする声道音響流パルスこのシーケンスから新興興奮させるし、鼻1 から放射口からと (ある程度) に結果としての音.生成された音のスペクトル構成主、声帯振動、喉頭のバイオメカニクスと2気管気流との相互作用によって支配の質によって影響されます。臨床と研究のコンテキストの両方で、ドキュメントと声帯振動の評価は、何よりも興味の発声を学ぶとき。
ヒトでは、体内の健全な生産の間に喉頭の直接内視鏡調査は挑戦的なとは現在技術的な手段を与えられるヒト以外の哺乳類のほぼ不可能です。したがって、慎重に制御保証物理的および/または生理学的な実験的境界条件、摘出 larynges3,4の使用する順番は、多くの場合の調査のための適切な置換体内音声生成機構。
声帯振動は、複雑な 3次元現象5です。従来の調査法のような (体内) 喉頭内視鏡検査または摘出喉頭準備通常ビューを提供のみ優れた声帯の振動6、彼らはの完全な三次元解析を許可しません。声帯運動。特に、優れたビューで声帯の下 (尾) 余白は見えない振動サイクルの主要な部分の間に。これは劣る (尾) と (頭蓋) 縁声帯、声帯振動5中に通常見られる現象間の位相遅延が原因です。数理モデルから調査結果をバックアップするための直接経験的な証拠は乏しい、ジオメトリの知識と低い声の動きを折るエッジ7、およびこうして声門チャネル8,9のジオメトリ,10は喉頭・声帯組織・結果の荷重や圧力11,12間の相互作用の理解のために重要です。恒例の優れたビューから隠されている声帯振動の別の側面は、2 つの声帯間の接触の垂直 (caudo 頭蓋) 深さです。連絡先奥行き歌 (レジスタ「裏声」対「胸」)13,14で使うボーカル レジスタの潜在的な指標は、声帯ヒダの縦の厚さに関連しています。
ここで 1 つの半分は、いわゆる片側喉頭セットアップ従来 (フル) 摘出喉頭製剤の欠点を克服するために利用できる喉頭が削除されるため、残りの振動特性の評価が容易3 つの次元で声帯。驚いたことに、この 1960 年代15設定の導入、1993年16における概念の初期検証多くない所はこの有望な実験的アプローチと17,18 実験を実行しています。 ,,1920,21,22,23。これのための説明は、実行可能な片側喉頭準備の作成の難しさの発見可能性があります。従来の切除 (フル) 喉頭の準備は十分に文書化4が、このような詳細な手順がないまだ片側喉頭のセットアップを作成するため。したがって、このペーパー レッド ディアの標本から実験結果によって補われる確実に再現可能な片側喉頭セットアップを確立するためのチュートリアルを提供する目的です。
ヘミ喉頭セットアップは測定装置、高速などの「従来型」の摘出喉頭セットアップまたはサウンドの生成中または適切な喉頭の構造物の振動を適切に文書に他のイメージング技術と多くの機能を共有します。加熱、加湿空気の供給。これらの一般的なセットアップの考慮事項は、本章4国立センターの音声、音声24から技術的なレポートで詳細に説明します。この原稿の範囲を超えてこれらの指示の繰り返しになります。ここでは、ヘミ喉頭セットアップを生成するため特殊なディレクティブのみが掲載されています。

<table><tbody><tr><td>Surgical blades</td><td>Surgeon</td><td>Jai Surgical Ltd., New Delhi, India</td><td></td></tr><tr><td>Saw</td><td>Hand saw (Lux, 150 mm length)</td><td>Lux, Wermelskirchen, Germany</td><td></td></tr><tr><td>Thermometer</td><td>Testo 922</td><td>Testo Ltd., Hampshire, UK</td><td>K-type Probe, Operating temperature -20 to +50 &amp;deg;C</td></tr><tr><td>Autoclave bags</td><td>Autoclave bags</td><td>vwr.com, VWR International s.r.o., Stribrna Skalice, Czech republic</td><td></td></tr><tr><td>Conductive glass plates</td><td>Custom made</td><td>UPOL - Joint laboratory of Optics&lt;br/&gt; Trida 17. listopadu 50A, 772 07 Olomouc, the Czech Rep.</td><td></td></tr><tr><td>Fixative cream</td><td>Denture fixative cream</td><td>Blend-a-dent Natural</td><td></td></tr><tr><td>Prongs and fastening system</td><td>Customized Kanya Al eloxed profiles</td><td>Distributor: VISIMPEX a.s.. Seifertova 33, 750 02 Prerov, the Czech Rep.;</td><td>&amp;nbsp;Combination of Kanya RVS and PVS fastening systems (http://www.kanya.cz/) + custom made prongs</td></tr><tr><td>Mounting tube</td><td>Custom made</td><td>UPOL - Joint laboratory of Optics,&lt;br/&gt; Trida 17. listopadu 50A, 772 07 Olomouc, the Czech Rep.</td><td></td></tr><tr><td>LED Light</td><td>Verbatim 52204 LED Lamp</td><td>Mitsubishi Chemical Holdings Corporation, Tokyo, Japan</td><td></td></tr><tr><td>Camera</td><td>Canon EOS1100D</td><td>Canon Inc.</td><td>18-55 mm lens</td></tr><tr><td>Airpump</td><td>Resun LP100</td><td>Resun</td><td></td></tr><tr><td>Strobe light</td><td>ELMED Helio-Strob micro2</td><td>ELMED Dr. Ing. Mense GmbH, Heiligenhaus, Germany</td><td></td></tr><tr><td>Humidifier</td><td>Custom made</td><td>Voice Research Lab, Dept. Biophysics, Faculty of Sciences, Palacky University Olomouc, Czech republic</td><td></td></tr><tr><td>Subglottic tract</td><td>Custom made adjustable subglottic tract</td><td>Voice Research Lab, Dept. Biophysics, Faculty of Sciences, Palacky University Olomouc, Czech republic</td><td>Hampala, V., Svec, Jan, Schovanek, P., and Mandat, D. Uzitny vzor c. 25585: Model subglotickeho traktu. [Utility model no. 25585: Model of subglottal tract] (In Czech) Soukup, P. 2013-27834(CZ 25505 U1), 1-7. 24-6-2013. Praha, Urad prumysloveho vlastnictvi</td></tr></tbody></table>

hemi-laryngeal setup for studying vocal fold vibration in three dimensions

ほとんどのヒト以外の哺乳類と人間の声は、声帯の自立振動を通して喉頭で生成されます。声帯振動の直接の映像記録は、ヒト以外の哺乳類で特に困難です。別の方法としては、摘出喉頭実験は管理された生理学的および物理的な条件の下で声帯振動を調査する機会を提供します。ただし、完全喉頭の使用は単空気力との相互作用観測による振動の構造の重要な部分を除く、声帯の平面図を提供します。ここで 1 つの半分にヘミ喉頭セットアップを利用してこの制限を克服できる喉頭の半ば矢削除、自励発振時に優れたと残りの声帯の横方向ビューを提供します。
ここでは、研究室のベンチに片側喉頭の構造と、取り付けの解剖学的な準備のためのステップ バイ ステップ ガイドが与えられます。ヘミ喉頭準備の模範的な発声は、2 つの同期カメラ (スーペリアーと横方向の眺め)、示す三次元声帯運動および対応する接触面積の時間変化を取り込んだ高速ビデオ データを記載されています。このパブリケーションでヘミ喉頭セットアップのドキュメント アプリケーションと再現性の実験的研究、発声のバイオメカニクスを理解する可能性のある音声科学者を提供することで容易になります。

ヘミ喉頭準備をし、前のセクションで参照している、空気供給管に取付部のイラストは、それぞれ図 1と図 2で提供されます。
2 つのカメラ アングルから声帯振動のドキュメント
片側喉頭の声帯の気流力自立振動上から...

Watch this Scientific Journal Video about Hemi-laryngeal Setup for Studying Vocal Fold Vibration in Three Dimensions at JoVE.com

Hemi-laryngeal Setup for Studying Vocal Fold Vibration in Three Dimensions

本稿ではヒトとヒト以外の哺乳類における発声の様々 な生物物理学的側面を調査するために声帯振動の多次元ビューを促進するヘミ喉頭標本の準備のためのプロトコル。

3 つの次元で声帯振動を勉強のため片側喉頭セットアップ

The voice of humans and most non-human mammals is generated in the larynx through self-sustaining oscillation of the vocal folds. Direct visual ...

hemi-laryngeal-setup-for-studying-vocal-fold-vibration-three

Research

JoVE Journal

Biology

10.9K ビュー.  Palacky University Olomouc. 本稿ではヒトとヒト以外の哺乳類における発声の様々 な生物物理学的側面を調査するために声帯振動の多次元ビューを促進するヘミ喉頭標本の準備のためのプロトコル。

ビデオ: Hemi-laryngeal Setup for Studying Vocal Fold Vibration in Three Dimensions

Studying Aggression in Drosophila (fruit flies)

Aggression is an innate behavior that evolved in the framework of defending or obtaining resources. This complex social behavior is influenced by genetic, hormonal and environmental factors. In many organisms, aggression is critical to survival but controlling and suppressing aggression in distinct contexts also has become increasingly important. In recent years, invertebrates have become increasingly useful as model systems for investigating the genetic and systems biological basis of complex social behavior. This is in part due to the diverse repertoire of behaviors exhibited by these organisms. In the accompanying video, we outline a method for analyzing aggression in Drosophila whose design encompasses important eco-ethological constraints. Details include steps for: making a fighting chamber; isolating and painting flies; adding flies to the fight chamber; and video taping fights. This approach is currently being used to identify candidate genes important in aggression and in elaborating the neuronal circuitry that underlies the output of aggression and other social behaviors.

Studying Aggression in Drosophila fruit flies

ショウジョウバエの勉強攻撃（ミバエ類）

Aggression is an innate behavior that evolved in the framework of defending or obtaining resources.  This complex social behavior is influenced by ...

生物学

Synthetic, Multi-Layer, Self-Oscillating Vocal Fold Model Fabrication

Sound for the human voice is produced via flow-induced vocal fold vibration. The vocal folds consist of several layers of tissue, each with differing material properties 1. Normal voice production relies on healthy tissue and vocal folds, and occurs as a result of complex coupling between aerodynamic, structural dynamic, and acoustic physical phenomena. Voice disorders affect up to 7.5 million annually in the United States alone 2 and often result in significant financial, social, and other quality-of-life difficulties. Understanding the physics of voice production has the potential to significantly benefit voice care, including clinical prevention, diagnosis, and treatment of voice disorders.
Existing methods for studying voice production include in vivo experimentation using human and animal subjects, in vitro experimentation using excised larynges and synthetic models, and computational modeling. Owing to hazardous and difficult instrument access, in vivo experiments are severely limited in scope. Excised larynx experiments have the benefit of anatomical and some physiological realism, but parametric studies involving geometric and material property variables are limited. Further, they are typically only able to be vibrated for relatively short periods of time (typically on the order of minutes).
Overcoming some of the limitations of excised larynx experiments, synthetic vocal fold models are emerging as a complementary tool for studying voice production. Synthetic models can be fabricated with systematic changes to geometry and material properties, allowing for the study of healthy and unhealthy human phonatory aerodynamics, structural dynamics, and acoustics. For example, they have been used to study left-right vocal fold asymmetry 3,4, clinical instrument development 5, laryngeal aerodynamics 6-9, vocal fold contact pressure 10, and subglottal acoustics 11 (a more comprehensive list can be found in Kniesburges et al. 12)
Existing synthetic vocal fold models, however, have either been homogenous (one-layer models) or have been fabricated using two materials of differing stiffness (two-layer models). This approach does not allow for representation of the actual multi-layer structure of the human vocal folds 1 that plays a central role in governing vocal fold flow-induced vibratory response. Consequently, one- and two-layer synthetic vocal fold models have exhibited disadvantages 3,6,8 such as higher onset pressures than what are typical for human phonation (onset pressure is the minimum lung pressure required to initiate vibration), unnaturally large inferior-superior motion, and lack of a "mucosal wave" (a vertically-traveling wave that is characteristic of healthy human vocal fold vibration).
In this paper, fabrication of a model with multiple layers of differing material properties is described. The model layers simulate the multi-layer structure of the human vocal folds, including epithelium, superficial lamina propria (SLP), intermediate and deep lamina propria (i.e., ligament; a fiber is included for anterior-posterior stiffness), and muscle (i.e., body) layers 1. Results are included that show that the model exhibits improved vibratory characteristics over prior one- and two-layer synthetic models, including onset pressure closer to human onset pressure, reduced inferior-superior motion, and evidence of a mucosal wave.

The methodology for fabricating synthetic vocal fold models is described. The models are life-sized and mimic the multi-layer structure of the human vocal folds. Results show the models to self-oscillate at pressures comparable to lung pressure and demonstrate flow-induced vibratory responses that are similar to those of human vocal folds.

合成、マルチレイヤ、自己発振声帯モデルの作製

Sound for the human voice is produced via flow-induced vocal fold vibration. The vocal folds consist of several layers of tissue, each with differing ...

生物工学

Chip-based Three-dimensional Cell Culture in Perfused Micro-bioreactors

We have developed a chip-based cell culture system for the three-dimensional cultivation of cells. The chip is typically manufactured from non-biodegradable polymers, e.g., polycarbonate or polymethyl methacrylate by micro injection molding, micro hot embossing or micro thermoforming. But, it can also be manufactured from bio-degradable polymers. Its overall dimensions are 0.7 1 x 20 x 20 x 0.7 1 mm (h x w x l). The main features of the chips used are either a grid of up to 1156 cubic micro-containers (cf-chip) each the size of 120-300 x 300 x 300 &mu; (h x w x l) or round recesses with diameters of 300 &mu; and a depth of 300 &mu; (r-chip). The scaffold can house 10 Mio. cells in a three-dimensional configuration. For an optimal nutrient and gas supply, the chip is inserted in a bioreactor housing. The bioreactor is part of a closed steril circulation loop that, in the simplest configuration, is additionaly comprised of a roller pump and a medium reservoir with a gas supply. The bioreactor can be run in perfusion, superfusion, or even a mixed operation mode. We have successfully cultivated cell lines as well as primary cells over periods of several weeks. For rat primary liver cells we could show a preservation of organotypic functions for more than 2 weeks. For hepatocellular carcinoma cell lines we could show the induction of liver specific genes not or only slightly expressed in standard monolayer culture. The system might also be useful as a stem cell cultivation system since first differentiation experiments with stem cell lines were promising.

We describe a chip-based platform for the three-dimensional cultivation of cells in micro-bioreactors. One chip can house up to 10 Mio. cells that can be cultivated under precisely defined conditions with regard to fluid flow, oxygen tension etc. in a sterile, closed circulation loop.

灌流マイクロバイオリアクターのチップベースの三次元細胞培養

We have developed a chip-based cell culture system for the three-dimensional cultivation of cells. The chip is typically manufactured from ...

Investigating the Three-dimensional Flow Separation Induced by a Model Vocal Fold Polyp

The fluid-structure energy exchange process for normal speech has been studied extensively, but it is not well understood for pathological conditions. Polyps and nodules, which are geometric abnormalities that form on the medial surface of the vocal folds, can disrupt vocal fold dynamics and thus can have devastating consequences on a patient&#39;s ability to communicate. Our laboratory has reported particle image velocimetry (PIV) measurements, within an investigation of a model polyp located on the medial surface of an in&#160;vitro driven vocal fold model, which show that such a geometric abnormality considerably disrupts the glottal jet behavior. This flow field adjustment is a likely reason for the severe degradation of the vocal quality in patients with polyps. A more complete understanding of the formation and propagation of vortical structures from a geometric protuberance, such as a vocal fold polyp, and the resulting influence on the aerodynamic loadings that drive the vocal fold dynamics, is necessary for advancing the treatment of this pathological condition. The present investigation concerns the three-dimensional flow separation induced by a wall-mounted prolate hemispheroid with a 2:1 aspect ratio in cross flow, i.e. a model vocal fold polyp, using an oil-film visualization technique. Unsteady, three-dimensional flow separation and its impact of the wall pressure loading are examined using skin friction line visualization and wall pressure measurements.

Vocal fold polyps can disrupt vocal fold dynamics and thus can have devastating consequences on a patient&#39;s ability to communicate. Three-dimensional flow separation induced by a wall-mounted model polyp and its impact on the wall pressure loading are examined using particle image velocimetry, skin friction line visualization, and wall pressure measurements.

モデルの声帯ポリープによる三次元流分離の調査

The fluid-structure energy exchange process for normal speech has been studied extensively, but it is not well understood for pathological conditions. ...

Construction and Characterization of a Novel Vocal Fold Bioreactor

In vitro engineering of mechanically active tissues requires the presentation of physiologically relevant mechanical conditions to cultured cells. To emulate the dynamic environment of vocal folds, a novel vocal fold bioreactor capable of producing vibratory stimulations at fundamental phonation frequencies is constructed and characterized. The device is composed of a function generator, a power amplifier, a speaker selector and parallel vibration chambers. Individual vibration chambers are created by sandwiching a custom-made silicone membrane between a pair of acrylic blocks. The silicone membrane not only serves as the bottom of the chamber but also provides a mechanism for securing the cell-laden scaffold. Vibration signals, generated by a speaker mounted underneath the bottom acrylic block, are transmitted to the membrane aerodynamically by the oscillating air. Eight identical vibration modules, fixed on two stationary metal bars, are housed in an anti-humidity chamber for long-term operation in a cell culture incubator. The vibration characteristics of the vocal fold bioreactor are analyzed non-destructively using a Laser Doppler Vibrometer (LDV). The utility of the dynamic culture device is demonstrated by culturing cellular constructs in the presence of 200-Hz sinusoidal vibrations with a mid-membrane displacement of 40 &#181;m. Mesenchymal stem cells cultured in the bioreactor respond to the vibratory signals by altering the synthesis and degradation of vocal fold-relevant, extracellular matrix components. The novel bioreactor system presented herein offers an excellent in vitro platform for studying vibration-induced mechanotransduction and for the engineering of functional vocal fold tissues.

A novel vocal fold bioreactor capable of delivering physiologically relevant, vibratory stimulation to cultured cells is constructed and characterized. This dynamic culture device, when combined with a fibrous poly(&#949;-caprolactone) scaffold, creates a vocal fold-mimetic environment that modulates the behaviors of mesenchymal stem cells.

新規声帯バイオリアクターの構築とキャラクタリゼーション

In vitro engineering of mechanically active tissues requires the presentation of physiologically relevant mechanical conditions to cultured cells. To ...

Microinjection Techniques for Studying Mitosis in the Drosophila melanogaster Syncytial Embryo

This protocol describes the use of the Drosophila melanogaster syncytial embryo for studying mitosis1. Drosophila has useful genetics with a sequenced genome, and it can be easily maintained and manipulated. Many mitotic mutants exist, and transgenic flies expressing functional fluorescently (e.g. GFP) - tagged mitotic proteins have been and are being generated. Targeted gene expression is possible using the GAL4/UAS system2. 

The Drosophila early embryo carries out multiple mitoses very rapidly (cell cycle duration, &asymp;10 min). It is well suited for imaging mitosis, because during cycles 10-13, nuclei divide rapidly and synchronously without intervening cytokinesis at the surface of the embryo in a single monolayer just underneath the cortex. These rapidly dividing nuclei probably use the same mitotic machinery as other cells, but they are optimized for speed; the checkpoint is generally believed to not be stringent, allowing the study of mitotic proteins whose absence would cause cell cycle arrest in cells with a strong checkpoint. Embryos expressing GFP labeled proteins or microinjected with fluorescently labeled proteins can be easily imaged to follow live dynamics (Fig. 1). In addition, embryos can be microinjected with function-blocking antibodies or inhibitors of specific proteins to study the effect of the loss or perturbation of their function3. These reagents can diffuse throughout the embryo, reaching many spindles to produce a gradient of concentration of inhibitor, which in turn results in a gradient of defects comparable to an allelic series of mutants. Ideally, if the target protein is fluorescently labeled, the gradient of inhibition can be directly visualized4. It is assumed that the strongest phenotype is comparable to the null phenotype, although it is hard to formally exclude the possibility that the antibodies may have dominant effects in rare instances, so rigorous controls and cautious interpretation must be applied. Further away from the injection site, protein function is only partially lost allowing other functions of the target protein to become evident.

Microinjection Techniques for Studying Mitosis in the Drosophila melanogaster Syncytial Embryo

This protocol describes the use of microinjection and high resolution imaging in the Drosophila melanogaster syncytial embryo to study mitosis.

に有糸分裂研究のためのマイクロインジェクション技術キイロショウジョウバエ合胞体胚

This protocol describes the use of the Drosophila melanogaster syncytial embryo  for studying mitosis1.  Drosophila has useful genetics with a sequenced ...

多層シリコーン声帯モデルの3Dプリンティング

Manufacturing Process for Non-Adhesive Super-Soft Vocal Fold Models

This study aims to develop super-soft, non-sticky vocal fold models for voice research. The conventional manufacturing process of silicone-based vocal fold models results in models with undesirable properties, such as stickiness and reproducibility issues. Those vocal fold models are prone to rapid aging, leading to poor comparability across different measurements. In this study, we propose a modification to the manufacturing process by changing the order of layering the silicone material, which leads to the production of non-sticky and highly consistent vocal fold models. We also compare a model produced using this method with a conventionally manufactured vocal fold model that is adversely affected by its sticky surface. We detail the manufacturing process and characterize the properties of the models for potential applications. The outcomes of the study demonstrate the efficacy of the modified fabrication method, highlighting the superior qualities of our non-sticky vocal fold models.&#160;The findings contribute to the development of realistic and reliable vocal fold models for research and clinical applications.

著者スポットライト:音声およびロボットアプリケーションのための合成声帯モデルの製造における進歩

This study demonstrates the manufacturing of non-sticky and super-soft vocal fold models by introducing a specific way to create the vocal fold layers, providing a detailed description of the manufacturing procedure, and characterizing the properties of the models.

非粘着性超軟質ボーカルフォールドモデルの製造工程

This study aims to develop super-soft, non-sticky vocal fold models for voice research. The conventional manufacturing process of silicone-based vocal ...

工学

Patch Clamp and Perfusion Techniques for Studying Ion Channels Expressed in Xenopus oocytes

The protocol presented here is designed to study the activation of the large conductance, voltage- and Ca2+-activated K+ (BK) channels. The protocol may also be used to study the structure-function relationship for other ion channels and neurotransmitter receptors1. BK channels are widely expressed in different tissues and have been implicated in many physiological functions, including regulation of smooth muscle contraction, frequency tuning of inner hair cells and regulation of neurotransmitter release2-6. BK channels are activated by membrane depolarization and by intracellular Ca2+ and Mg2+ 6-9. Therefore, the protocol is designed to control both the membrane voltage and the intracellular solution. In this protocol, messenger RNA of BK channels is injected into Xenopus laevis oocytes (stage V-VI) followed by 2-5 days of incubation at 18&deg;C10-13. Membrane patches that contain single or multiple BK channels are excised with the inside-out configuration using patch clamp techniques10-13. The intracellular side of the patch is perfused with desired solutions during recording so that the channel activation under different conditions can be examined. To summarize, the mRNA of BK channels is injected into Xenopus laevis oocytes to express channel proteins on the oocyte membrane; patch clamp techniques are used to record currents flowing through the channels under controlled voltage and intracellular solutions.

Patch Clamp and Perfusion Techniques for Studying Ion Channels Expressed in Xenopus oocytes

Ionic current of BK channels is recorded using patch clamp techniques. BK channels are expressed in Xenopus oocytes by injecting messenger RNA. The intracellular solution during patch clamp recordings is controlled by a perfusion system.

イオンチャネル研究のためのパッチクランプおよび灌流のテクニックで表さアフリカツメガエル卵母細胞

The protocol presented here is designed to study the activation of the large conductance, voltage- and Ca2+-activated K+ (BK) channels. The protocol may ...

3 つの次元で声帯振動を勉強のため片側喉頭セットアップ

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ショウジョウバエの勉強攻撃（ミバエ類）

合成、マルチレイヤ、自己発振声帯モデルの作製

灌流マイクロバイオリアクターのチップベースの三次元細胞培養

モデルの声帯ポリープによる三次元流分離の調査

新規声帯バイオリアクターの構築とキャラクタリゼーション

に有糸分裂研究のためのマイクロインジェクション技術キイロショウジョウバエ合胞体胚

非粘着性超軟質ボーカルフォールドモデルの製造工程

非粘着性超軟質ボーカルフォールドモデルの製造工程

非粘着性超軟質ボーカルフォールドモデルの製造工程

イオンチャネル研究のためのパッチクランプおよび灌流のテクニックで表さアフリカツメガエル卵母細胞