この作品は、アメリカ耳鼻科学会からCAMへの助成金によって、大学病院事件医療センターからKNAとCAMへの助成金によって部分的にサポートされていました。我々は、顕微鏡カメラで私たちを提供するためのフォーカスメディカルグループ株式会社（ヒンクリー、オハイオ州）でジェフChurneyに感謝したいと思います。ビデオ制作サービスは、我々は、ビデオの準備中に彼らのコメントのためにCWRUでジョナサンフランケルとアレックスレヴィットに感謝融合クリエイティブマーケティング株式会社（ヴァルドスタ、ジョージア州）の好意により提供。

パート1：麻酔と手術の準備<ol><li>成人では、チャールズリバーインターナショナルラボラトリーズ社（ウィルミントン、MA）からメスダンカン- Hartley系モルモットにケタミンを60 mg / kgおよびキシラジン12 mg / kgの筋肉内で麻酔です。 </li><li>石油の潤滑剤は、乾燥による角膜損傷から動物を保護するために目に適用されます。 </li><li>動物は、肩に頭の上からと耳の間に剃毛されています。 </li><li>剃毛面積を7.5％のエチルアルコール70％続いてポビドンヨードを持つ最初の滅菌方法で整形処理されています。 </li><li>動物は、加温ブランケットと手術台の上に、頭と曲がる首を上昇プラットフォーム上に配置されます。動物の頭部は、外科医から指示される。首の十分な屈曲が十分に後頭隆起を公開するために必要です。 </li><li>動物は無菌的に掛けている。 </li></ol>パート2：骨の後頭部の切開と露出<ol><li>ブピバカインに加えてエピネフリン（Marcaine）0.25％は将来の切開部位（後頭部に沿って正中背部皮膚の切開）に沿って適用されます。これは、出血を制限する、ローカルの血管収縮を引き起こし、動物が目覚めの後に持続的な術後鎮痛を提供します。 </li><li>正中背部皮膚の切開は、後頭部に沿って頭皮に行われます。 <img src="/files/ftp_upload/1728/1728fig1.jpg" alt="図1" /> 図1：上記のような皮膚の切開は、背側正中線に沿って行われます。 </li><li>適切な露出を得るために、皮膚は切開の両側に小さな曲線の止血剤を置くことによって横方向に収納されています。 </li><li>水平方向の切開は、骨の後頭部を公開するために首の筋肉に沿って行われます。この段階では動物が大後頭神経の切除のために適切な麻酔にもかかわらず、ピクッと動くことができる。脊髄切除は後頭下部に水平切開と危険です。この合併症は、下層の骨が常に手術用メスの刃の下に感じられるようにすることで回避することができます。 </li><li>骨に切開後頭下筋群と、第三止血剤は、優れた筋肉の端を撤回するために使用されます。この止血剤は、外科の助手によって固定されています。 </li><li> Lempert骨膜起子は、側頭後頭縫合糸を公開するために使用されます。 </li></ol>パート3：側頭後頭縫合を掘削<ol><li>頭頂後頭縫合（図2）、1.5ミリメートルダイヤモンドバリでotologicドリルの露光時に縫合するinferio -横方向にドリルダウンするために使用されます。 </li><li>頭蓋骨の外側のテーブルの下に、板間層（頭蓋骨のテーブルの間に介在する海綿骨）の海綿状のテクスチャが見えるようになります。板間層が離れて掘削されると、頭蓋骨の内側の表は、S状静脈洞である淡い青色の色調を持つと共に、表示されます。 </li><li> S状静脈洞のマージンは、容器を貫通することなく定義されています。 </li><li> S状静脈洞を覆う骨が十分に薄くされると、より高い倍率が利用されるとストレートピックは慎重にS状静脈洞を覆う骨を除去するために使用されます。 </li></ol><img src="/files/ftp_upload/1728/1728fig2.jpg" alt="図2" /> 図2は、部分的に掘削される領域は、側頭後頭縫合糸を覆う。 第4部：管閉塞<ol><li>スケルトンS状静脈洞は、蓋を明らかにするために内側に引っ込められる。 </li><li> 0.5ミリメートルダイヤモンドバリは、蓋の内側と内リンパ嚢とダクトへのドリルダウンするために使用されます。 </li><li>内リンパ管は、まっすぐなピックを使用して骨ワックスが詰まっています。 </li><li>頭蓋骨の欠損は、Gelfoam（ファイザー社、ニューヨーク、NY）で補強されています。 </li><li> 4から0 POLYSORB（米国外科、ノーウォーク、コネチカット州）縫合糸は、筋肉を結集するために使用されます。皮膚切開を閉じるための皮下縫合の技術へのシームレスな移行は、そこです。この閉鎖で、その後縫合糸を除去する必要はないと動物は、サイトでの爪と縫合糸を抜くなるおそれがないものであること。 </li></ol>第5部：術後ケアモルモットは、体温を維持するためにインキュベーターに入れて、目を覚ましと胸骨の位置を維持することが可能になるまで30分間隔でチェックされます。動物は、呼吸速度とボディの色のために検査されます。このセンターは、それぞれ、術後感染の予防と痛みのコントロールのためにエンロフロキサシン（皮下22.7 mg / mLに、0.2 mL）およびブプレノルフィン（皮下0.3mg/mL 0.05 mLを、）を使用します。鎮痛は術後一日1と2で一度手術の日に二回与えられます。抗生物質は、無菌のsuのために必要なものではありませんが、rgery、予防的抗生物質5日間、1日2回のコースでは、術後3日目givenonce可能性があります。 合併症と管理<ol><li>呼吸抑制は、外科医は、常に動物の呼吸に注意する必要があります。呼吸が減速または停止した場合、手術を中止し、視覚的に胸部の動きを点検。動物が呼吸困難を経験したり、苦しい呼吸している場合、口と気道が閉塞されていないことを確認してください。 </li><li>出血が停止するまでS状静脈洞への浸透は、吸引チップと一緒にGelfoamと圧縮を使用してください。 </li><li>硬膜浸透は、Gelfoamの欠陥をカバー。 </li><li>前庭障害は、S状静脈洞への横方向の1mm以上のドリルでください。モルモットのすべてのデータが無効になる後半規管を、傷つけるようなリスクをして。 </li></ol>

<ol>
	<li>Andrews, J. C., Bohmer, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+surgical+approach+to+the+endolymphatic+sac+and+the+cochlear+aqueduct+in+the+guinea+pig.">The surgical approach to the endolymphatic sac and the cochlear aqueduct in the guinea pig.</a> Am J Otolaryngol. 10, 61-66 (1989).</li><li>Kimura, R., Schuknecht, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Membranous+hydrops+in+the+inner+ear+of+the+guinea+pig+after+obliteration+of+the+endolymphatic+sac.">Membranous hydrops in the inner ear of the guinea pig after obliteration of the endolymphatic sac.</a> Pract Otolaryngol. , (1965).</li></ol>

動物実験は、ケースウェスタンリザーブ大学の動物実験使用の委員会が定めるガイドラインおよび規則に従って行った。

内リンパ管の外科的閉塞後の数週間では、動物対象は聴覚誘発脳幹反応によって測定されるように変動する感音難聴を経験する。聴性脳幹反応（ABR）の記録のサンプルを図3に示します。左耳は逃れたコントロールとして機能します。 <a href="https://www.jove.com/files/ftp_upload/1728/1728fig4.jpg"><img border="0" src="/files/ftp_upload/1728/1728fig3tmb.jpg" alt="図3" /></a> 図3：モルモット28週間後の...

surgical induction of endolymphatic hydrops by obliteration of the endolymphatic duct

内リンパ管の閉塞や閉塞によるモルモットの内リンパ水腫の外科的誘導（ELH）が条件と人間のメニエール病のために相互に関連付けることが重要、よく受け入れられた動物モデルである。 1965年に、ロバート木村とハロルドSchuknechtは、最初の内リンパ管の閉塞（木村1965）のための硬膜内のアプローチを説明した。有効であるが、脳の保護カバーの浸透を必要とするこの手法は、、動物の被験者における罹患率と死亡率の望ましくないレベルを計上した。その結果、アンドリュースとベーマーは予想中枢神経系（CNS）浸透。（アンドリュースとベーマー1989）に関連する合併症の少ないを生成する硬膜外アプローチを開発し、 硬膜外アプローチは、ここで説明した最初の基本的な筋層を露出させる後頭部の地域における正中切開を必要とします。我々は右側のみで動作します。上を覆う組織の適切な収縮した後、水平方向の切開は、右頭頂後頭縫合線を公開する権利後頭部の筋肉に行われます。 S状静脈洞が見えるようになるまで、骨はすぐにinferio横ずれ縫合線（図1）その後otologicドリルで掘削された。 S状静脈洞の内側陥凹は、内リンパ嚢を収容する内リンパ管の蓋を明らかにする。内リンパ嚢の領域に蓋の内側の掘削は、閉塞し、最終的にELHを生成するために骨蝋を充填した内リンパ管を、明らかに。 次の週では、動物はプログレッシブ、変動する難聴とELHの組織学的証拠を紹介します。

Watch this Scientific Journal Video about Surgical Induction of Endolymphatic Hydrops by Obliteration of the Endolymphatic Duct at JoVE.com

Surgical Induction of Endolymphatic Hydrops by Obliteration of the Endolymphatic Duct

このビデオでは、外科的内リンパ水腫を生成するためにモルモットの内リンパ管を妨害する方法を示しています。

内リンパ管の閉塞による内リンパ水腫の外科的誘導

Surgical induction of endolymphatic hydrops (ELH) in the guinea pig by obliteration and obstruction of the endolymphatic duct is a well-accepted animal ...

surgical-induction-endolymphatic-hydrops-obliteration-endolymphatic

Research

JoVE Journal

Biology

11.3K ビュー.  Case Western Reserve University. このビデオでは、外科的内リンパ水腫を生成するためにモルモットの内リンパ管を妨害する方法を示しています。

ビデオ: Surgical Induction of Endolymphatic Hydrops by Obliteration of the Endolymphatic Duct

Imaging Effector Memory T cells in the Ear After Induction of Adoptive DTH

Delayed type hypersensitivity (DTH) is an immune reaction in which the main players are CCR7- effector / memory T lymphocytes. Here, we demonstrate a method for inducing and recording the progress of a DTH reaction in the rat ear. This is followed by a demonstration of the preparation of rat ear tissue for two-photon imaging of the CCR7- effector / memory T cell response.
An adoptive DTH is induced by the intraperitoneal injection of GFP-labeled Ova-specific CCR7- effector / memory T cell line (Beeton, C J. Visualized Experiments, Issue 8). Cells are then allowed to equilibrate in the rat for 48 hours before challenge by injecting one ear with saline (control ear) and the other with a 1:1 mix of Ova and Ova conjugated to Texas-Red (Ova-TR) to allow visualization of resident antigen-presenting cells.
We describe a method of tissue preparation useful for imaging the motility of cells within the deep dermal layer during an immune response, in conjunction with visualization of collagen fibers by second harmonic generation. Ear tissue is cut into 5 x 5 mm squares (slightly larger is better) and mounted onto plastic cover slips using Vetbond&trade;, which are then secured using silicone grease in an imaging chamber and superfused by oxygen-bubbled tissue culture medium at 37&deg;C.

Here we demonstrate a method for inducing and recording the progress of a delayed type-hypersensitivity (DTH) reaction in the rat ear. This is followed by a demonstration of the preparation of rat ear tissue for two-photon imaging of the effector / memory T cell response.

養子DTHの誘導後の耳のイメージングエフェクターメモリーT細胞

Delayed type hypersensitivity (DTH) is an immune reaction in which the main players are CCR7- effector / memory T lymphocytes. Here, we demonstrate a ...

生物学

Assay for Neural Induction in the Chick Embryo

The chick embryo is a valuable tool in the study of early embryonic development. Its transparency, accessibility and ease of manipulation, make it an ideal tool for studying the formation and initial patterning of the nervous system. This video demonstrates how to graft organizer tissue into a host, a method by which Hensen s node (the organizer in the chick embryo) is grafted to a host competent ectoderm. The organizer graft instructs overlying na ve tissue to adopt a neural fate via neural inducing signals. This mechanism is referred to as neural induction, and constitutes the initial step in the formation of brain and spinal cord in amniotes. This method is essentially used for the characterization of putative neural inducing molecules in chick. This video demonstrates the different steps in the assay for neural induction; First, the donnor embryo is explanted and pinned on a dish. Then, the host embryo is prepared for New culture. The graft is excised and transplanted to the host area pellucida margin. The host is cultured for 18-22 hrs. The assembly is fixed and processed for further applications (e.g. in situ hybridization). This method was originally devised by Waddington 1,2 and Gallera 3,4.

Neural induction is the first step in the formation of the brain. It is a mechanism by which Hensen's node (organizer), instructs adjacent tissue to adopt a neural fate, i.e. to give rise to the nervous system. This video demonstrates an assay for neural induction in chick embryo.

鶏胚における神経誘導のためのアッセイ

The chick embryo is a valuable tool in the study of early embryonic development. Its transparency, accessibility and ease of manipulation, make it an ...

Microsurgical Clip Obliteration of Middle Cerebral Aneurysm Using Intraoperative Flow Assessment

Cerebral aneurysms are abnormal widening or ballooning of a localized segment of an intracranial blood vessel.  Surgical clipping is an important treatment for aneurysms which attempts to exclude blood from flowing into the aneurysmal segment of the vessel while preserving blood flow in a normal fashion.  Improper clip placement may result in residual aneurysm with the potential for subsequent aneurysm rupture or partial or full occlusion of distal arteries resulting in cerebral infarction.  Here we describe the use of an ultrasonic flow probe to provide quantitative evaluation of arterial flow before and after microsurgical clip placement at the base of a middle cerebral artery aneurysm.  This information helps ensure adequate aneurysm reconstruction with preservation of normal distal blood flow.

Description of the surgical obliteration of a cerebral aneurysm utilizing an ultrasonic flow probe to assess arterial flow prior to and after aneurysm clip placement.

術中フローアセスメントを用いた中大脳動脈瘤の顕微クリップ閉塞

Cerebral aneurysms are abnormal widening or ballooning of a localized segment of an intracranial blood vessel.  Surgical clipping is an important ...

Isolation of Basal Cells and Submucosal Gland Duct Cells from Mouse Trachea

The large airways are directly in contact with the environment and therefore susceptible to injury from toxins and infectious agents that we breath in 1. The large airways therefore require an efficient repair mechanism to protect our bodies. This repair process occurs from stem cells in the airways and isolating these stem cells from the airways is important for understanding the mechanisms of repair and regeneration. It is also important for understanding abnormal repair that can lead to airway diseases 2. The goal of this method is to isolate a novel stem cell population from the mouse tracheal submucosal gland ducts and to place these cells in in vitro and in vivo model systems to identify the mechanisms of repair and regeneration of the submucosal glands 3. This production shows methods that can be used to isolate and assay the duct and basal stem cells from the large airways 3.This will allow us to study diseases of the airway, such as cystic fibrosis, asthma and chronic obstructive pulmonary disease. Currently, there are no methods for isolation of submucosal gland duct cells and there are no in vivo models to study the regeneration of submucosal glands.

Here we demonstrate our protocol for isolation of basal and submucosal gland duct cells from mouse tracheas. We also demonstrate the method of injecting stem cells into the dorsal mouse fat pad to create an in vivo model of submucosal gland regeneration.

マウス気管から基底細胞および粘膜下腺管細胞の単離

The large airways are directly in contact with the environment and therefore susceptible to injury from toxins and infectious agents that we breath in 1. ...

Culturing Primary Rat Inner Medullary Collecting Duct Cells

Arginine-vasopressin (AVP) facilitates water reabsorption by renal collecting duct principal cells and thereby fine-tunes body water homeostasis. AVP binds to vasopressin V2 receptors (V2R) on the surface of the cells and thereby induces synthesis of cAMP. This stimulates cellular signaling processes leading to changes in the phosphorylation of the water channel aquaporin-2 (AQP2). Protein kinase A phoshorylates AQP2 and thereby triggers the translocation of AQP2 from intracellular vesicles into the plasma membrane facilitating water reabsorption from primary urine. Aberrations of AVP release from the pituitary or AVP-activated signaling in principal cells can cause central or nephrogenic diabetes insipidus, respectively; an elevated blood plasma AVP level is associated with cardiovascular diseases such as chronic heart failure and the syndrome of inappropriate antidiuretic hormone secretion.
Here, we present a protocol for cultivation of primary rat inner medullary collecting duct (IMCD) cells, which express V2R and AQP2 endogenously. The cells are suitable for elucidating molecular mechanisms underlying the control of AQP2 and thus to discover novel drug targets for the treatment of diseases associated with dysregulation of AVP-mediated water reabsorption. IMCD cells are obtained from rat renal inner medullae and are used for experiments six to eight days after seeding. IMCD cells can be cultured in regular cell culture dishes, flasks and micro-titer plates of different formats, the procedure only requires a few hours, and is appropriate for standard cell culture laboratories.

Arginine-vasopressin (AVP) controls fine-tuning of body water homeostasis through facilitating water reabsorption by renal principal cells. Here, we present a protocol for the cultivation of primary rat inner medullary collecting duct cells suitable for the elucidation of molecular mechanisms underlying AVP-mediated water reabsorption.

管細胞の収集培養初代ラットインナー髄

Arginine-vasopressin (AVP) facilitates water reabsorption by renal collecting duct principal cells and thereby fine-tunes body water homeostasis. AVP ...

Induction and Analysis of Epithelial to Mesenchymal Transition

Epithelial to mesenchymal transition (EMT) is essential for proper morphogenesis during development. Misregulation of this process has been implicated as a key event in fibrosis and the progression of carcinomas to a metastatic state. Understanding the processes that underlie EMT is imperative for the early diagnosis and clinical control of these disease states. Reliable induction of EMT in vitro is a useful tool for drug discovery as well as to identify common gene expression signatures for diagnostic purposes. Here we demonstrate a straightforward method for the induction of EMT in a variety of cell types. Methods for the analysis of cells pre- and post-EMT induction by immunocytochemistry are also included. Additionally, we demonstrate the effectiveness of this method through antibody-based array analysis and migration/invasion assays.

A straightforward method is described for the induction of epithelial to mesenchymal transition (EMT) in a variety of cell types. Methods for the analysis of cells in EMT states by immunocytochemistry are included.

間葉転換を誘導し、上皮の分析

Epithelial  to mesenchymal transition (EMT) is essential for proper morphogenesis during  development. Misregulation of this  process has been implicated ...

Isolation of Human Hepatocytes by a Two-step Collagenase Perfusion Procedure

The liver, an organ with an exceptional regeneration capacity, carries out a wide range of functions, such as detoxification, metabolism and homeostasis. As such, hepatocytes are an important model for a large variety of research questions. In particular, the use of human hepatocytes is especially important in the fields of pharmacokinetics, toxicology, liver regeneration and translational research. Thus, this method presents a modified version of a two-step collagenase perfusion procedure to isolate hepatocytes as described by Seglen 1.
Previously, hepatocytes have been isolated by mechanical methods. However, enzymatic methods have been shown to be superior as hepatocytes retain their structural integrity and function after isolation. This method presented here adapts the method designed previously for rat livers to human liver pieces and results in a large yield of hepatocytes with a viability of 77&plusmn;10%. The main difference in this procedure is the process of cannulization of the blood vessels. Further, the method described here can also be applied to livers from other species with comparable liver or blood vessel sizes.

A modified two-step collagenase perfusion procedure for isolation of human hepatocytes is described. This method can also be applied to other mammalian livers. The isolated hepatocytes are available in high yield and viability, making them a suitable model for scientific research in areas such as liver regeneration, pharmacokinetics and toxicology.

二段階コラゲナーゼ灌流手順によって、ヒト肝細胞の分離

The liver, an organ with an exceptional regeneration capacity, carries out a wide range of functions, such as detoxification, metabolism and homeostasis. ...

In Vivo 4-Dimensional Tracking of Hematopoietic Stem and Progenitor Cells in Adult Mouse Calvarial Bone Marrow

Through a delicate balance between quiescence and proliferation, self renewal and production of differentiated progeny, hematopoietic stem cells (HSCs) maintain the turnover of all mature blood cell lineages. The coordination of the complex signals leading to specific HSC fates relies upon the interaction between HSCs and the intricate bone marrow microenvironment, which is still poorly understood[1-2].
We describe how by combining a newly developed specimen holder for stable animal positioning with multi-step confocal and two-photon in vivo imaging techniques, it is possible to obtain high-resolution 3D stacks containing HSPCs and their surrounding niches and to monitor them over time through multi-point time-lapse imaging. High definition imaging allows detecting ex vivo labeled hematopoietic stem and progenitor cells (HSPCs) residing within the bone marrow. Moreover, multi-point time-lapse 3D imaging, obtained with faster acquisition settings, provides accurate information about HSPC movement and the reciprocal interactions between HSPCs and stroma cells.
Tracking of HSPCs in relation to GFP positive osteoblastic cells is shown as an exemplary application of this method. This technique can be utilized to track any appropriately labeled hematopoietic or stromal cell of interest within the mouse calvarium bone marrow space.

In Vivo 4-Dimensional Tracking of Hematopoietic Stem and Progenitor Cells in Adult Mouse Calvarial Bone Marrow

The nature of the interactions between hematopoietic stem and progenitor cells (HSPCs) and bone marrow niches is poorly understood. Custom hardware modifications and a multi-step acquisition protocol allow the use of two-photon and confocal microscopy to image ex vivo labeled HSPCs homed within bone marrow areas, tracking interactions and movement.

成体マウス頭蓋冠骨髄中の造血幹細胞および前駆細胞のin vivo 4次元トラッキングで

Through a delicate balance between quiescence and proliferation, self renewal and production of differentiated progeny, hematopoietic stem cells (HSCs) ...

Fecal Glucocorticoid Analysis: Non-invasive Adrenal Monitoring in Equids

Adrenal activity can be assessed in the equine species by analysis of feces for corticosterone metabolites. During a potentially aversive situation, corticotrophin releasing hormone (CRH) is released from the hypothalamus in the brain. This stimulates the release of adrenocorticotrophic hormone (ACTH) from the pituitary gland, which in turn stimulates release of glucocorticoids from the adrenal gland. In horses the glucocorticoid corticosterone is responsible for several adaptations needed to support equine flight behaviour and subsequent removal from the aversive situation. Corticosterone metabolites can be detected in the feces of horses and assessment offers a non-invasive option to evaluate long term patterns of adrenal activity. Fecal assessment offers advantages over other techniques that monitor adrenal activity including blood plasma and saliva analysis. The non-invasive nature of the method avoids sampling stress which can confound results. It also allows the opportunity for repeated sampling over time and is ideal for studies in free ranging horses. This protocol describes the enzyme linked immunoassay (EIA) used to assess feces for corticosterone, in addition to the associated biochemical validation.

Adrenal activity can be assessed in the equine species by analysis of feces for corticosterone metabolites. The method offers a non-invasive option to assess long term patterns in both domestic and free ranging horses. This protocol describes the enzyme linked immunoassay involved and the associated biochemical validation.

糞便グルココルチコイド分析：ウマにおける非侵襲性副腎モニタリング

Adrenal activity can be assessed in the equine species by analysis of feces for corticosterone metabolites. During a potentially aversive situation, ...

Partial Bile Duct Ligation in the Mouse: A Controlled Model of Localized Obstructive Cholestasis

In rodents, complete bile duct ligation (cBDL) of the common bile duct is an established surgical technique for studying obstructive cholestasis and bile duct proliferation. However, long-term experiments can lead to increased morbidity and mortality. In select mouse strains with underlying liver disease, meaningful comparisons can be made even with ligation of a single lobe of the liver, which can reduce animal losses and expenses. Here, we describe partial bile duct ligation (pBDL) in the mouse, in which only the left hepatic bile duct is ligated, causing biliary obstruction in the left lobe but not the remaining lobes. With careful microsurgical technique, pBDL experiments can be cost-effective, since the unligated lobe serves as an internal control to the ligated lobes, when subjected to the same conditions in the same animal. Unlike cBDL, a separate sham-operated control group is not necessary. pBDL is highly useful to directly compare localized versus systemic effects of cholestasis and other retained bile components. pBDL can also be repurposed as a novel method to investigate mechanisms related to medications and cell migration.

Here, we present partial bile duct ligation as a surgical model of liver injury and regeneration in rodents.

マウスの部分胆管結紮: ローカライズされた閉塞性胆汁うっ滞の制御モデル

In rodents, complete bile duct ligation (cBDL) of the common bile duct is an established surgical technique for studying obstructive cholestasis and bile ...