この作品は、LAP（PI）への補助金によって支えられている。 国民健康R21 HD50558の研究所、2005から07を受賞、国立健康R01 HD045822の研究所は、2004から09を受賞。ダナ財団助成を、 イノベーションのためのカナダの財団（&quot;CFI&quot;助成金）、授与、2004から06を受賞 2008-2012、オンタリオの研究助成は、2008から2012を受賞。

パート1：参加者の前に研究室に到着<ol><li>部屋は参加者に邪魔になる可能性があります余分な記事がないことを保証すること。 </li><li> - セットアップとfNIRS日立ETG - 4000のシステムで実験的なプロトコルをロードする。 </li><li>あなたの実験パラダイムを設定します。実験的なパラダイムはEprime、プレゼンテーション、PsyscopeやMatlabベースの心理学のツールボックスを含む別のプレゼンテーションソフトウェアでプログラムすることができます。ここでは、MATLABベースの心理学のツールボックスを使用してください。 </li><li>タイミングは、このように実験的なパラダイムが完全にデータの収集とタイムアウトする必要がある、データ解析のためのキーです。 fNIRS日立ETG - 4000は、データ収集またはその逆をトリガするために実験的なパラダイムを可能にする、機能をトリガしている。 fNIRS日立ETG - 4000のプレゼンテーションプログラムのトリガテストします。トリガは、並列、逐次、またはUSBポートを使用して行うことができます。ここでは、パラレルポートを介してトリガを示す。 </li><li>前のfNIRSの調査を開始するには、参加者の背景のスクリーニングを行うことが重要です。プティト教授の研究室では、我々は参加者や親が勉強、適切な標準化されたアンケートを29に記入することによって背景のスクリーニングを実施する。 </li></ol>参加者が到着<ol start="6"><li>セッションを実施するために、専門の方法で参加者を治療することが重要です。実験が始まる前に、参加者または参加者の両親/法的保護者が同意書に署名する必要があります。それは、これらの重要でエキサイティングな実験では彼らの時間のために参加者に感謝することが重要です。 </li><li>参加者は快適に近いfNIRSの試験室に装着されている。乳児の参加者は、親の膝の上に取り付けられていない可能性があります。 </li></ol>パート2：Optodesを配置＆10月20日システムを使用する一貫性のあるデータの解釈を可能にする分析法の別のコンポーネントには、fNIRS記録プロトコルの標準化です。これはoptode配置、参加者の位置、および刺激提示ソフトウェアのトリガを伴います。プローブの正確な神経解剖学的配置の両方と関心領域（ROI）の確認、10〜20システム3,4,30を使用して実現されています。さらに、プローブの配列の定位局在は、各プローブの位置3,4に配置されたビタミンEのカプセルを実施した参加者の解剖学的MRI共同登録のスキャンにPolhemus社の高速TRAKシステムから3Dトラッキングの情報を重ねることによって参加者の頭蓋骨に確認された。最適な参加者の位置は、本体または椅子に接触することなく緩く吊り下げ、光ファイバと、リクライニングチェアで快適に、参加者を置くことに関与。 <ol><li>以下のヘッドの測定は、巻尺で撮影し、参加者のデータシートを下に書かれている。 <ul><li>周りイニオンへ[後]頭骨</li><li>トップ上Inionsへ[後]頭骨</li><li>トップ上の耳への耳</li></ul></li><li>サージカルテープは、特定のターゲットの位置をマークするために使用することができます。この実験では、FP、T3/T4、F8/F7を迎えます</li><li> Optodeの配列は、実験の目的によって指示10月20日のポイントに停泊して特定のoptodesと参加者の頭の上に配置されています。 </li></ol>パート4：Optodeアレイのテスト<ol><li>日立ETG - 4000のGUIインタフェースとプローブテストの紹介。 </li><li>信号をテストする：一度optodesが参加者の頭皮に配置され、信号品質がテストされます。 optodeは明確なシグナルがない場合は、研究者がやさしくoptodeと頭皮の接続から毛を取り除く。機会にoptodesはア​​ルコール綿で拭くことが必要な場合があります。 </li></ol>パート5：実験を実行する。 <ol><li>少なくとも二つの実験者が部屋に常に存在する必要があります。fNIRS日立ETG - 4000リアルタイム読み出すと参加者を観察し、他を観察つ。参加者に焦点を当てたビデオカメラを持っておくことを高く事後観察のために推奨されます。 fNIRS日立ETG - 4000の利点は、ビデオとfNIRSの信号が同期し、共同で登録されていることです。生成されたすべての関連情報およびファイルを含むログが保持されます。 </li><li>実験的な血行動態のパラダイムを構築するのよく確立されたメソッドが存在する、すなわち設計およびイベント関連のデザインをブロックします。より完全な説明については最近のレビュー論文31を参照してください。 </li></ol>パート6：分析すべてのデータが収集された、参加者は自分の時間と参加意欲のために感謝し、ラボを残している。分析がfNIRS日立ETG - 4000上で行われていないとして、として、ではなく、データは分析のコンピュータにエクスポートされます。 <ol><li>ヘモグロビン濃度へμVからの変換。生の減衰値はレーザー強度の減衰（のようなμVで測定）で収集されるように、これらの値は、酸素と脱酸素化ヘモグロビンVALUに変換する必要があります。ES。これは、修正されたランバート - ベールの法則を使って行われます。 </li><li>修正Beer - Lambertののアプリケーションは、2つの手順で行われている。散乱は、最初のパスの長さ、信号の計算されたベースラインへのそれぞれの波長（ΔAλ（t）は ）タスク（I タスク ）の間に光強度の光学濃度を比較することによって計算される（のための減衰にわたって一定であるという仮定の下で私ベースライン ）。各波長のΔAの値と変更されたランバート-ベールの法則を解決するためにサンプリングした時点（t）を。 <table border="0"><tbody><tr><td> 式（1） <img style="float:left;" src="/files/ftp_upload/1268/eq1.jpg" alt="式（1）" /></td><td> 式（2） <img style="float:left;" src="/files/ftp_upload/1268/eq2.jpg" alt="式（2）" /></td></tr></tbody></table></li></ol> λ1∈デオキシ、λ1∈オキシ 、波長λ2∈デオキシとλ2∈酸素は、組織内の単位濃度の距離ごとに吸収するために失われた光の割合を測定する吸光係数の定数である。その結果、CのデオキシとC オキシ値は、各tに対して脱酸素化と酸素化ヘモグロビンの濃度です。 パート7：代表的な結果いくつかの明確な特性の典型的な血行動態の応答結果。酸素ヘモグロビンの応答では、特徴的なディップは、最初にあります。ニューロンの領域が活性化し、利用可能な酸素を枯渇させるようにこのディップが発生します。血流が増加するにつれて、酸素化ヘモグロビンを運ぶ、オキシヘモグロビンの応答は、定常状態のレベルに最初のベースラインレベルの上に急速に上昇する。領域はもはやアクティブにされていないときは、酸素ヘモグロビンの応答次の12から15秒以上の減衰ゆっくりとではベースラインレベルに下がります。初期のベースラインレベルに戻った血液動態反応の前に発生したアンダーシュートが時折あります。 悪い結果が適切に頭皮や過度の運動に装着されていないoptodesの形で通常です。ノイズのこれらのタイプ - &quot;きっぱりと&quot;、と呼ばれる - マイクロボルトの値が飽和し、異なるチャンネル数、状況に応じたやり方で酸素とデオキシ-応答の両方の動きとして、信号で顕著となっている。 デモ：光ファイバを振る。 統計的分析：各チャネルごとに、各参加者のと各タスクの抽出された酸素とデオキシヘモグロビンの値は、従来の統計分析に提出することができる、t -検定を含め、ANOVAs、相関等

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The authors have nothing to disclose.

本研究では、我々は人間の認知や知覚に関連して人間の脳機能を調べるために小説、非侵襲的fNIRSの脳のイメージング技術の使用を実証。 fNIRSを脳機能イメージングは​​、一日研究室で広く利用できる可能性があること、特に幼児や子供の集団で、医師のオフィスを非侵襲的脳機能イメージングの未来を表す場合があります、と臨床医がに関する基本的な科学的知見を適用できるように学?...

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exploring cognitive functions in babies, children &amp; adults with near infrared spectroscopy

そのような言語1,2,3,4,5,6,7,8,9,10、メモリ11、及び注意12などの高次認知過程に関連皮質の活性化を調査する赤外分光法（fNIRS）の研究、近機能の爆発進行中の典型的な非定型認知20,21,22と3,4,13,14,15,16,17,18,19世界的に関わる大人、子供や乳幼児です。認知神経科学のためfNIRSを用いての現代的課題は、彼らが23,24,25,26普遍的に解釈可能であり、したがって、機能的な組織と人間のより高い認知の基礎となる神経システムに関する重要な科学的問題を進めることができるようデータの体系的分析を達成することです。 既存のニューロイメージング技術は、どちらか少ない堅牢な時間や空間分解能を持っている。ポジトロンCTと機能的磁気共鳴イメージング（PETやfMRIが）より良い空間分解能を持っているのに対し、事象関連電位と磁気脳機能イメージング（ERPやMEG）は、優れた時間分解能を持っている。オキシヘモグロビンは優先的に680nmのとデオキシヘモグロビンによって吸収される近赤外域（700〜1000 nm）を、で、光の非イオン化の波長を使用しては、優先的に830nmの（例えば、実際に、非常に波長がにハードワイヤードで吸収されるここに示したfNIRS日立ETG - 400システム）、fNIRSはよくそれは放射線と良好な空間分解能（〜4cmの深さ）を使用せずに良好な時間分解能（〜5秒）の両方を持っているので、より高い認知の研究に最適、とされないことです参加者は密閉構造27,28に存在する必要があります。快適に普通の椅子（大人、子供）に座って、あるいはお母さんのラップ（乳幼児）に座っている間皮質活動の参加者が評価できる。特に、NIRSは事実上サイレント、（デスクトップのコンピュータのサイズ）ユニークなポータブルであり、そして参加者の微妙な動きに耐えることができます。これは、必ずしもそのキーコンポーネントの一つ音声生成や手話の手の​​中に口の動きとして持っている人間の言語の神経研究の場合に特に顕著である。 血行動態応答がローカライズされている方法は、レーザーエミッターと検出器の配列によってです。検出器は、皮質表面から反射される量を検出しながらエミッターは、非電離光の既知の強度を発する。近くに一緒にoptodesを、より空間分解能、さらに離れてoptodes一方、浸透の大きい深さ。のためのfNIRS日立ETG - 4000システム最適普及/解像度optodeの配列は、2センチメートルに設定されています。 私たちの目標は、フィールドを標準化し、共通のバックグラウンドを持っている別のfNIRSのラボが世界中で有効にするためのfNIRSデータの取得と分析の手法を示すことです。

Watch this Scientific Journal Video about Exploring Cognitive Functions in Babies, Children & Adults with Near Infrared Spectroscopy at JoVE.com

Exploring Cognitive Functions in Babies, Children &amp; Adults with Near Infrared Spectroscopy

ここでは、特に子供の脳の発達を研究するには、赤外分光法（fNIRS）、認知神経科学で使用される新規非侵襲的脳機能イメージングシステムの近くに機能するためのデータ収集とデータ分析の方法を説明します。このメソッドは、データ収集およびデータの解釈と科学的発見に不可欠な分析の普遍的な標準を提供します。

近赤外分光法で赤ちゃん、子供と大人の認知機能を探る

An explosion of functional Near Infrared Spectroscopy (fNIRS) studies investigating cortical activation in relation to higher cognitive processes, such as ...

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20.4K ビュー.  University of Michigan, Ann Arbor. ここでは、特に子供の脳の発達を研究するには、赤外分光法（fNIRS）、認知神経科学で使用される新規非侵襲的脳機能イメージングシステムの近くに機能するためのデータ収集とデータ分析の方法を説明します。このメソッドは、データ収集およびデータの解釈と科学的発見に不可欠な分析の普遍的な標準を提供します。

ビデオ: Exploring Cognitive Functions in Babies, Children & Adults with Near Infrared Spectroscopy

Recordings of Neural Circuit Activation in Freely Behaving Animals

The relationship between patterns of neural activity and corresponding behavioral expression is difficult to establish in unrestrained animals. Traditional non-invasive methods require at least partially restrained research subjects, and they only allow identification of large numbers of simultaneously activated neurons. On the other hand, small ensembles of neurons or individual neurons can only be measured using single-cell recordings obtained from largely reduced preparations. Since the expression of natural behavior is limited in restrained and dissected animals, the underlying neural mechanisms that control such behavior are difficult to identify. 
Here, I present a non-invasive physiological technique that allows measuring neural circuit activation in freely behaving animals. Using a pair of wire electrodes inside a water-filled chamber, the bath electrodes record neural and muscular field potentials generated by juvenile crayfish during natural or experimentally evoked escape responses. The primary escape responses of crayfish are mediated by three different types of tail-flips which move the animals away from the point of stimulation. Each type of tail-flip is controlled by its own neural circuit; the two fastest and most powerful escape responses require activation of different sets of large &#8220;command&#8221; neurons. In combination with behavioral observations, the bath electrode recordings allow unambiguous identification of these neurons and the associated neural circuits. Thus activity of neural circuitry underlying naturally occurring behavior can be measured in unrestrained animals and in different behavioral contexts.

Non-invasive measurements of neural activity patterns in freely behaving animals are obtained by combining neurophysiological recordings with high speed videography.

自由に動物を振る舞うの神経回路のアクティベーションの記録

The relationship between patterns of neural activity and corresponding behavioral expression is difficult to establish in unrestrained animals. ...

生物学

Assembly, Tuning and Use of an Apertureless Near Field Infrared Microscope for Protein Imaging

This paper aims to instruct the reader in the assembly and operation of an infrared near-field microscope for imaging beyond the diffraction limit.  The apertureless near-field microscope is a light scattering-type instrument that provides infrared spectra at circa 20 nm resolution. A complete list of components and a step-by-step protocol for use is provided.  Common errors in assembly and instrument tuning are discussed.  A representative data set that shows the secondary structure of an amyloid fibril is presented.  

The assembly of a nearfield infrared microscope for imaging protein aggregates is described.

アセンブリは、チューニングとタンパク質イメージングのためApertureless近距離赤外線顕微鏡の使用

This paper aims to instruct the reader in the assembly and operation of an infrared near-field microscope for imaging beyond the diffraction limit.  The ...

Measurement Of Neuromagnetic Brain Function In Pre-school Children With Custom Sized MEG

Magnetoencephalography is a technique that detects magnetic fields associated with cortical activity [1]. The electrophysiological activity of the brain generates electric fields  - that can be recorded using electroencephalography (EEG)- and their concomitant magnetic fields  - detected by MEG. MEG signals are detected by specialized sensors known as superconducting quantum interference devices (SQUIDs). Superconducting sensors require cooling with liquid helium at -270 &deg;C. They are contained inside a vacumm-insulated helmet called a dewar, which is filled with liquid. SQUIDS are placed in fixed positions inside the helmet dewar in the helium coolant, and a subject's head is placed inside the helmet dewar for MEG measurements. The helmet dewar must be sized to satisfy opposing constraints. Clearly, it must be large enough to fit most or all of the heads in the population that will be studied. However, the helmet must also be small enough to keep most of the SQUID sensors within range of the tiny cerebral fields that they are to measure. Conventional whole-head MEG systems are designed to accommodate more than 90% of adult heads. However adult systems are not well suited for measuring brain function in pre-school chidren whose heads have a radius several cm smaller than adults. The KIT-Macquarie Brain Research Laboratory at Macquarie University uses a MEG system custom sized to fit the heads of pre-school children. This child system has 64 first-order axial gradiometers with a 50 mm baseline[2] and is contained inside a magnetically-shielded room (MSR) together with a conventional adult-sized MEG system [3,4]. There are three main advantages of the customized helmet dewar for studying children. First, the smaller radius of the sensor configuration brings the SQUID sensors into range of the neuromagnetic signals of children's heads. Second, the smaller helmet allows full insertion of a child's head into the dewar. Full insertion is prevented in adult dewar helmets because of the smaller crown to shoulder distance in children. These two factors are fundamental in recording brain activity using MEG because neuromagnetic signals attenuate rapidly with distance. Third, the customized child helmet aids in the symmetric positioning of the head and limits the freedom of movement of the child's head within the dewar. When used with a protocol that aligns the requirements of data collection with the motivational and behavioral capacities of children, these features significantly facilitate setup, positioning, and measurement of MEG signals.

The advent of MEG systems sized for young children opens important new opportunities to study brain development. The new system, together with a protocol that aligns experimental requirements with the capacities of children, can be used to study cognitive and language processes in healthy, awake children aged three to six.

カスタムサイズのMEGと就学前のお子様でNeuromagnetic脳機能の計測

Magnetoencephalography is a technique that detects magnetic fields associated with cortical activity [1]. The electrophysiological activity of the brain ...

Method for the Assessment of Effects of a Range of Wavelengths and Intensities of Red/near-infrared Light Therapy on Oxidative Stress In Vitro

Red/near-infrared light therapy (R/NIR-LT), delivered by laser or light emitting diode (LED), improves functional and morphological outcomes in a range of central nervous system injuries in vivo, possibly by reducing oxidative stress. However, effects of R/NIR-LT on oxidative stress have been shown to vary depending on wavelength or intensity of irradiation. Studies comparing treatment parameters are lacking, due to absence of commercially available devices that deliver multiple wavelengths or intensities, suitable for high through-put in vitro optimization studies. This protocol describes a technique for delivery of light at a range of wavelengths and intensities to optimize therapeutic doses required for a given injury model. We hypothesized that a method of delivering light, in which wavelength and intensity parameters could easily be altered, could facilitate determination of an optimal dose of R/NIR-LT for reducing reactive oxygen species (ROS) in vitro.
Non-coherent Xenon light was filtered through narrow-band interference filters to deliver varying wavelengths (center wavelengths of 440, 550, 670 and 810nm) and fluences (8.5 x 10-3 to 3.8 x 10-1 J/cm2) of light to cultured cells. Light output from the apparatus was calibrated to emit therapeutically relevant, equal quantal doses of light at each wavelength. Reactive species were detected in glutamate stressed cells treated with the light, using DCFH-DA and H2O2 sensitive fluorescent dyes.&#160;
We successfully delivered light at a range of physiologically and therapeutically relevant wavelengths and intensities, to cultured cells exposed to glutamate as a model of CNS injury. While the fluences of R/NIR-LT used in the current study did not exert an effect on ROS generated by the cultured cells, the method of light delivery is applicable to other systems including isolated mitochondria or more physiologically relevant organotypic slice culture models, and could be used to assess effects on a range of outcome measures of oxidative metabolism.

Method for the Assessment of Effects of a Range of Wavelengths and Intensities of Red/near-infrared Light Therapy on Oxidative Stress In Vitro

Non-coherent Xenon light was passed through narrow-band interference and neutral density filters to deliver light of varying wavelength and intensity to cultured cells. This protocol was used to assess the effects of red/near-infrared light therapy on production of reactive species in vitro: no effects were observed using the tested parameters.

酸化ストレスに赤/近赤外光療法の波長および強度の範囲の効果の評価のための手法<em&gt;インビトロ</em

Red/near-infrared light therapy (R/NIR-LT), delivered by laser or light emitting diode (LED), improves functional and morphological outcomes in a range of ...

Near Infrared (NIr) Light Increases Expression of a Marker of Mitochondrial Function in the Mouse Vestibular Sensory Epithelium

Strategies for attenuating decline in balance function with increasing age are predominantly focused on physical therapies including balance tasks and exercise. However, these approaches do not address the underlying causes of balance decline. Using mice, the impact of near infrared light (NIr) on the metabolism of cells in the vestibular sensory epithelium was assessed. Data collected shows that this simple and safe intervention may protect these vulnerable cells from the deleterious effects of natural aging. mRNA was extracted from the isolated peripheral vestibular sensory epithelium (crista ampullaris and utricular macula) and subsequently transcribed into a cDNA library. This library was then probed for the expression of ubiquitous antioxidant (SOD-1). Antioxidant gene expression was then used to quantify cellular metabolism. Using transcranial delivery of NIr in young (4 weeks) and older (8 - 9 months) mice, and a brief treatment regime (90 sec/day for 5 days), this work suggests NIr alone may be sufficient to improve mitochondrial function in the vestibular sensory epithelium. Since there are currently no available, affordable, non-invasive methods of therapy to improve vestibular hair cell function, the application of external NIr radiation provides a potential strategy to counteract the impact of aging on cellular metabolism inthe vestibular sensory epithelium.

Near Infrared NIr Light Increases Expression of a Marker of Mitochondrial Function in the Mouse Vestibular Sensory Epithelium

Mitochondrial dysfunction is a hallmark of cellular senescence. This paper uses non-invasive near-infrared (NIr) treatment to improve mitochondrial function in the aging mouse vestibular sensory epithelium.

近赤外（NIR）の光は、マウス前庭感覚上皮におけるミトコンドリア機能のマーカーの発現を増加させる

Strategies for attenuating decline in balance function with increasing age are predominantly focused on physical therapies including balance tasks and ...

Functional Reconstitution and Channel Activity Measurements of Purified Wildtype and Mutant CFTR Protein

The Cystic Fibrosis Transmembrane Conductance Regulator (CFTR) is a unique channel-forming member of the ATP Binding Cassette (ABC) superfamily of transporters. The phosphorylation and nucleotide dependent chloride channel activity of CFTR has been frequently studied in whole cell systems and as single channels in excised membrane patches. Many Cystic Fibrosis-causing mutations have been shown to alter this activity. While a small number of purification protocols have been published, a fast reconstitution method that retains channel activity and a suitable method for studying population channel activity in a purified system have been lacking. Here rapid methods are described for purification and functional reconstitution of the full-length CFTR protein into proteoliposomes of defined lipid composition that retains activity as a regulated halide channel. This reconstitution method together with a novel flux-based assay of channel activity is a suitable system for studying the population channel properties of wild type CFTR and the disease-causing mutants F508del- and G551D-CFTR. Specifically, the method has utility in studying the direct effects of phosphorylation, nucleotides and small molecules such as potentiators and inhibitors on CFTR channel activity. The methods are also amenable to the study of other membrane channels/transporters for anionic substrates.

Described here is a rapid and effective procedure for functional reconstitution of purified wild-type and mutant CFTR protein that preserves activity for this chloride channel, which is defective in Cystic Fibrosis. Iodide efflux from reconstituted proteoliposomes mediated by CFTR allows studies of channel activity and the effects of small molecules.

精製された野生型および変異型CFTRタンパク質の機能的再構成とチャネル活性の測定

The Cystic Fibrosis Transmembrane Conductance Regulator (CFTR) is a unique channel-forming member of the ATP Binding Cassette (ABC) superfamily of ...

The ChIP-exo Method: Identifying Protein-DNA Interactions with Near Base Pair Precision

Chromatin immunoprecipitation (ChIP) is an indispensable tool in the fields of epigenetics and gene regulation that isolates specific protein-DNA interactions. ChIP coupled to high throughput sequencing (ChIP-seq) is commonly used to determine the genomic location of proteins that interact with chromatin. However, ChIP-seq is hampered by relatively low mapping resolution of several hundred base pairs and high background signal. The ChIP-exo method is a refined version of ChIP-seq that substantially improves upon both resolution and noise. The key distinction of the ChIP-exo methodology is the incorporation of lambda exonuclease digestion in the library preparation workflow to effectively footprint the left and right 5&#39; DNA borders of the protein-DNA crosslink site. The ChIP-exo libraries are then subjected to high throughput sequencing. The resulting data can be leveraged to provide unique and ultra-high resolution insights into the functional organization of the genome. Here, we describe the ChIP-exo method that we have optimized and streamlined for mammalian systems and next-generation sequencing-by-synthesis platform.

Here, we present a protocol to achieve near base pair resolution of protein-DNA interactions. This is obtained by exonuclease treatment of DNA fragments selectively enriched by chromatin immunoprecipitation (ChIP-exo) followed by high throughput sequencing.

ChIP-エキソ方法：近く塩基対の精度でタンパク質 -  DNA相互作用を同定します

Chromatin immunoprecipitation (ChIP) is an indispensable tool in the fields of epigenetics and gene regulation that isolates specific protein-DNA ...

Production of Near-Infrared Sensitive, Core-Shell Vaccine Delivery Platform

Vaccine delivery strategies that can limit the exposure of cargo to organic solvent while enabling novel release profiles are crucial for improving immunization coverage worldwide. Here, a novel injectable, ultraviolet- curable and delayed burst release- enabling vaccine delivery platform called polybubbles is introduced. Cargo was injected into polyester-based polybubbles that were formed in 10% carboxymethycellulose -based aqueous solution. This paper includes protocols to maintain spherical shape of the polybubbles and optimize cargo placement and retention to maximize the amount of cargo within the polybubbles. To ensure safety, chlorinated solvent content within the polybubbles were analyzed using neutron activation analysis. Release studies were conducted with small molecules as cargo within the polybubble to confirm delayed burst release. To further show the potential for on-demand delivery of the cargo, gold nanorods were mixed within the polymer shell to enable near-infrared laser activation.

This article describes the protocols used to produce a novel vaccine delivery platform, &#34;polybubbles,&#34; to enable delayed burst release. Polyesters including poly(lactic-co-glycolic acid) and polycaprolactone were used to form the polybubbles and small molecules and antigen were used as cargo.

近赤外感受性コアシェルワクチンデリバリープラットフォームの製造

Vaccine delivery strategies that can limit the exposure of cargo to organic solvent while enabling novel release profiles are crucial for improving ...

Exploring Adipose Tissue Structure by Methylsalicylate Clearing and 3D Imaging

Obesity is a major worldwide public health issue that increases the risk to develop cardiovascular diseases, type-2 diabetes, and liver diseases. Obesity is characterized by an increase in adipose tissue (AT) mass due to adipocyte hyperplasia and/or hypertrophia, leading to profound remodeling of its three-dimensional structure. Indeed, the maximal capacity of AT to expand during obesity is pivotal to the development of obesity-associated pathologies. This AT expansion is an important homeostatic mechanism to enable adaptation to an excess of energy intake and to avoid deleterious lipid spillover to other metabolic organs, such as muscle and liver. Therefore, understanding the structural remodeling that leads to the failure of AT expansion is a fundamental question with high clinical applicability. In this article, we describe a simple and fast clearing method that is routinely used in our laboratory to explore the morphology of mouse and human white adipose tissue by fluorescent imaging. This optimized AT clearing method is easily performed in any standard laboratory equipped with a chemical hood, a temperature-controlled orbital shaker and a fluorescent microscope. Moreover, the chemical compounds used are readily available. Importantly, this method allows one to resolve the 3D AT structure by staining various markers to specifically visualize the adipocytes, the neuronal and vascular networks, and the innate and adaptive immune cells distribution.

Here, we describe a simple, inexpensive and fast clearing method to resolve the 3D structure of both mouse and human white adipose tissue using a combination of markers to visualize vasculature, nuclei, immune cells, neurons, and lipid-droplet coat proteins by fluorescent imaging.

メチルサリチル酸除去と3Dイメージングによる脂肪組織構造の探索

Obesity is a major worldwide public health issue that increases the risk to develop cardiovascular diseases, type-2 diabetes, and liver diseases. Obesity ...

Dissecting Mechanoenzymatic Properties of Processive Myosins with Ultrafast Force-Clamp Spectroscopy

Ultrafast force-clamp spectroscopy (UFFCS) is a single molecule technique based on laser tweezers that allows the investigation of the chemomechanics of both conventional and unconventional myosins under load with unprecedented time resolution. In particular, the possibility to probe myosin motors under constant force right after the actin-myosin bond formation, together with the high rate of the force feedback (200 kHz), has shown UFFCS to be a valuable tool to study the load dependence of fast dynamics such as the myosin working stroke. Moreover, UFFCS enables the study of how processive and non-processive myosin-actin interactions are influenced by the intensity and direction of the applied force.
By following this protocol, it will be possible to perform ultrafast force-clamp experiments on processive myosin-5 motors and on a variety of unconventional myosins. By some adjustments, the protocol could also be easily extended to the study of other classes of processive motors such as kinesins and dyneins. The protocol includes all the necessary steps, from the setup of the experimental apparatus to sample preparation, calibration procedures, data acquisition and analysis.

Presented here is a comprehensive protocol to perform ultrafast force-clamp experiments on processive myosin-5 motors, which could be easily extended to the study of other classes of processive motors. The protocol details all the necessary steps, from the setup of the experimental apparatus to sample preparation, data acquisition and analysis.

超高速フォースクランプ分光法によるミオシンのメカノ酵素的性質の解剖

Ultrafast force-clamp spectroscopy (UFFCS) is a single molecule technique based on laser tweezers that allows the investigation of the chemomechanics of ...