この原稿の数値の一部は寛大バートO.ウィリアムスの研究室によって提供されました。著者は、国立癌研究所によってサポートされていU54CA143803、CA163124、CA093900、およびCA143055を付与します。

このプロトコルは、ジョンズ・ホプキンス大学施設内動物管理使用委員会によって設定されたガイドラインを満たし、次の。 1.マイクロ解剖マウス前立腺<ol><li>二酸化炭素窒息または施設の動物の管理と使用のガイドラインに従った代替承認の方法により、マウスを安楽死させます。 </li><li> 4足のそれぞれを通ってピンを置くことによって、解剖ボードに仰臥位でマウスをピン。 </li><li> 70％エタノールで腹部をスプレーします。 </li><li>鉗子で下腹部の皮膚を持ち、皮膚を介してカットし、ペニスの開口部に約1cmの前方を腹膜するはさみを使用して、任意の臓器を切らないように注意してください。 </li><li>腹腔を露出させ、アップ腹部の両側胸郭に、下腹部から皮膚及び腹膜を切り取ります。 </li><li>膀胱と尿生殖路を特定し、優しく側に脂肪および他の器官を動かすことによって、それらを公開。 </li><li> USIngの鉗子、グリップ尿道に近いベースでの輸精管を、そしてそれを離れて引き裂きます。 </li><li>反対輸精管で繰り返します。 </li><li>同時に膀胱下の尿道と腹側前立腺（膀胱のベース下の約1cm）を切断するはさみを使用しながら、慎重にグリップ、膀胱を鉗子を使用して、それを引き上げます。 注：膀胱がプルアップされているように、（膀胱、尿道のセクション、すべての前立腺葉、および精嚢を含む）全体の尿生殖路は、それが付属します。多少の抵抗があるかもしれません。 </li><li>約5〜10 mLのリン酸緩衝生理食塩水（PBS）を含む60〜100ミリメートルペトリ皿に尿生殖路（UGT）を配置します。 </li><li>解剖顕微鏡下でプロトコルの残りの部分を実行します。 2細かい鉗子、それぞれの手で1を使用してください。 「鉛筆のような」鉗子を持ち、それらを安定するように、作業台の上に腕を休ませます。 </li><li>膀胱があるようUGTを位置決めするために微細なピンセットを使用して上、尿道が下向きされ、精嚢は、膀胱のいずれかの側に配置されています。 </li><li>グリップ1鉗子、慎重に任意の前立腺組織を離れて引き裂くことなく、他の鉗子で脂肪を引き離すと尿道。注：脂肪は周囲の組織に比べて「ピカピカ」が表示されます。 </li><li>脂肪が除去されると、前立腺の2腹葉間結合組織を引き裂くとそれらを分離するために鉗子を使用しています。 </li><li> 、腹側ローブの1を削除グリップに先端付近の単一腹葉を1鉗子を使用し、可能な限り尿道に近いベースでそのローブグリップに他の鉗子を使用します。まだ葉の基部を把持しながら、グリップに尿道を他の鉗子を使用します。しっかりと、滑らかな動きで離れて尿道から葉を引き出します。何の前立腺組織は尿道に添付残っていないことを確認してください。次の実験段階に応じて、適切な培地に葉を置きます。 <ol><li> Sを固定し、パラフィンに埋め込むには組織学的分析のための十分な、10％中性緩衝ホルマリン（NBF）8に葉を配置します。 </li><li>凍結保存のために、このような10月のように、メディアを凍結し、8を区画に葉を置きます。 </li><li>培養のために単一細胞を単離するための冷PBSで葉を置くか9フローサイトメトリー。 </li></ol></li><li>他の腹葉と繰り返し手順1.15。 </li><li>必ず精嚢を穿刺しないようになって、静かに精嚢から葉を引っ張るために鉗子を使用して、前葉の1を削除します。ローブは精嚢から独立していると、ステップ1.15で腹葉と同じ方法でそれを削除します。 </li><li>他の前方ローブと繰り返し手順1.17。 </li><li>背側前立腺が表示されていることなどの上に残存尿生殖路を反転します。 </li><li> 2背外側ローブともローブとtとの結合組織の間の結合組織を引き裂くために鉗子を使用し、背外側ローブの1を削除するには彼は精嚢領域を後方。ステップ1.15で腹葉と同様の方法で葉を削除します。 </li><li>他の背外側葉と繰り返し手順1.20。 </li></ol> 2.去勢手術注：これは生存の手術なので、無菌状態、麻酔、および疼痛管理は、このプロトコルの倫理的、正常に完了することが重要です。すべての機関の動物の管理と使用のガイドラインに従ってください。低体温症を防ぐために、外科手術の際に、このような再循環水ブランケットとして、外部熱源を使用してください。それは胸骨横臥位を維持するのに十分な意識を取り戻したまで無人の動物を放置しないでください。完全に回復するまで、他の動物の会社に手術を受けた動物を返さないでください。手術で使用する前に、すべての手術器具をオートクレーブ。 <ol><li>清潔、無菌の表面に適切な措置（ 例えば 2％吸入イソフルラン）を用いてマウスを麻酔し、メートルを配置仰臥位でウーズ。注：例えば、ケタミンのような代替的な麻酔法を、承認された動物プロトコルに応じて、また使用され得ます。 </li><li>つま先をつまんで、マウスの反射神経をチェックしてください。マウスが反応しない場合は、次のステップに進みます。それが反応しない場合は、1-3分待ってから再度確認してください。電気かみそりで手術領域を剃ります。 </li><li>無菌的に交互に70％エタノールおよびヨウ素のスクラブや制度獣医学のスタッフやIACUCによって推奨される手順を用いてマウスの腹部を準備します。 </li><li>滅菌メスを使用して、下腹部の正中線に皮膚を介して陰茎に約1.5センチ前方を1センチメートル縦切開を行います。 </li><li>腹膜を介して小切開（&lt;1センチ）ことを確認し、任意の臓器を切らないように注意してください。 </li><li>滅菌ピンセット、グリップ腹膜のカットエッジを使用し、下に腹膜腔を見ることができるように静かに持ち上げます。 </li><li>別の滅菌ピンセットを使用して、目に到達電子膀胱に対する横腹腔とグリップ左右いずれかの睾丸脂肪パッド、。精巣は、同様に出てくるまで、腹膜および皮膚内の開口部を介して脂肪パッドを引き出します。このステップの間に、他の組織や臓器を傷つけないように注意してください。 </li><li>焼灼ペンを使用して、精巣を保持している脂肪パッドを切断。焼灼ペンノートと精巣動脈を通ってゆっくりとカット：精巣動脈が速すぎてカットされている場合、そこにあまりにも多くの出血であってもよく、マウスを安楽死させなければならないことがあります。 </li><li>脂肪パッドと動脈が切断されると、精巣および脂肪パッドを削除します。 </li><li>繰り返して、他の精巣で2.6から2.9を繰り返します。注：精巣を除去するための別の方法は、はさみで血管を切断し、その後、精巣動脈を外科的に結紮することです。 </li><li>腹膜は、吸収性縫合糸で閉じ縫合糸。 </li><li>ステープル皮膚は、外科的創傷クリップで閉じました。 </li><li>痛みを管理するための鎮痛剤を投与し、そして場所5-10分間暖かいケージのクリーンケージにマウス。痛みや出血の兆候についてマウスを監視します。 </li></ol>

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著者は、彼らが競合する金融利害関係を持たないことを宣言します。

前立腺顕微解剖は、マウス前立腺（ 図1）の葉特有の実験および分析を可能にします。遺伝子操作マウスモデルにおいて、表現型は、他で見られないもので葉に見られます。また、組織学的分析のために、このプロトコルは、純粋な前立腺組織の最大量は節中に存在する他の尿生殖路組織せず、切片と染色することができることを保証します。最後に、単一細胞実験のために、こ...

前立腺は、米国における男性の癌の最も一般的な部位です。ほぼ22万男性毎年前立腺癌と診断され、約27,000人の男性は、自分の病気1に屈するます。男性は米国1における前立腺癌の診断の1 7の生涯リスクを持っています。良性前立腺肥大（BPH）、前立腺の年齢関連非癌性拡大は、80 2以上の男性の80％に影響を及ぼし、また広範な条件である。このように、前立腺はかなりの研究の焦点です。 マウスモデルは、前立腺3の疾患を研究するために広く用いられています。全体的に、マウスの前立腺は、ヒト腺4の優れた代表であるが、マウスとヒトの前立腺解剖学と生理学5の類似点と相違点があります。両種では、前立腺が尿道を取り囲む、膀胱の基部に位置しています。ヒト前立腺腺は、単一のLOです中央、トランジション、周辺機器、および前部線維筋間質：4つのゾーンに分け、こと。前側、腹、および背外側：これとは対照的に、マウスの前立腺は尿道の周りの異なる位置に3対になった葉で構成されています。 1、それは1：マウス6で4細胞レベルでは、主な違いは、ヒトにおける細胞比率を管腔する基底細胞が約1であることです。マウス間質は、ヒトへの相対的なマウスでも異なっている - 筋層は、マウス前立腺における非常に薄いですしながら、人間は、より広範な平滑筋を持っています。マウス前立腺の適切な識別、解剖、およびハンドリングは、マウスの前立腺研究に不可欠です。 ホルモンのレベルが大幅にヒトとマウスの両方で前立腺の開発と恒常性に影響を与えます。エストロゲンは、前立腺の発達7において役割を果たすと思われるが、前立腺における最も重要なホルモンはテストステロンです。テストステロンは腺開発ANのために不可欠ですDのメンテナンス。物理的または化学的のいずれかに続いて去勢、成人前立腺apoptose内腔側細胞の約90％、および腺が収縮します。テストステロンを去勢条件下で個体に再導入された場合、前立腺全容量に自分自身を再生することができます。テストステロンはまた、アンドロゲン除去療法が一般的に使用される治療戦略である理由である、前立腺癌を駆動するように思われます。しかし、多くの前立腺癌は、アンドロゲン除去に耐性になります。また、前立腺癌の治療のためのアンドロゲン枯渇療法を受けている男性は去勢、再生条件のサイクルを受けます。マウスでは、外科的去勢は、テストステロンを再導入することにより、前立腺のホルモン再生と共に、去勢抵抗性前立腺に対するテストステロンの循環の影響を研究すると重要なツールです。 この記事では、我々が議論し、locatすることにより、適切な技術のデモンストレーションを行いますeおよびマイクロ解剖マウスの前立腺を、同様にマウスを去勢外科的します。

<table border="0" cellpadding="0" cellspacing="0" width="741">
	<tbody>
		<tr height="21">
			<td height="21" style="height:21px;width:244px;">Surgical tools</td>
			<td style="width:145px;">Roboz</td>
			<td style="width:113px;">Various</td>
			<td style="width:239px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Formaldehyde</td>
			<td>Sigma</td>
			<td>F8775</td>
			<td></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;">OCT medium</td>
			<td>VWR</td>
			<td>25608-930</td>
			<td style="width:239px;">Must be placed on dry ice to freeze, and stored at -80C</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">PBS</td>
			<td>Life Technologies</td>
			<td align="right">10010</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Isoflurane</td>
			<td>Johns Hopkins</td>
			<td></td>
			<td>Also available from vendors online</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Cautery Pen</td>
			<td>Medline</td>
			<td>ESCT002</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Silk suture</td>
			<td>AD Surgical</td>
			<td>M-S330R19</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Surgical Wound Clips</td>
			<td>Roboz</td>
			<td>RS-9265</td>
			<td></td>
		</tr>
	</tbody>
</table>

murine prostate micro-dissection and surgical castration

Mouse models are used extensively to study prostate cancer and other diseases. The mouse is an excellent model with which to study the prostate and has been used as a surrogate for discoveries in human prostate development and disease. Prostate micro-dissection allows consistent study of lobe-specific prostate anatomy, histology, and cellular characteristics in the absence of contamination of other tissues. Testosterone affects prostate development and disease. Androgen deprivation therapy is a common treatment for prostate cancer patients, but many prostate tumors become castration-resistant. Surgical castration of mouse models allows for the study of castration resistance and other facets of hormonal biology on the prostate. This procedure can be coupled with testosterone reintroduction, or hormonal regeneration of the prostate, a powerful method to study stem cell lineages in the prostate. Together, prostate micro-dissection and surgical castration opens up a multitude of opportunities for robust and consistent research of prostate development and disease. This manuscript describes the protocols for prostate micro-dissection and surgical castration in the laboratory mouse.

すべての6前立腺葉前立腺顕微解剖（ 図1）を介してマウスから除去しました。完全な尿生殖路（UGT）は、すべての前立腺葉、膀胱、精嚢、尿道（ 図1a）で構成されています。輸精管が尿道に付着するが、前立腺顕微解剖は不要であり、したがって、尿道（ 図1B-1C）を切断することによって、UGTの除去の前に取り外すことができま?...

Watch this Scientific Journal Video about Murine Prostate Micro-dissection and Surgical Castration at JoVE.com

Murine Prostate Micro-dissection and Surgical Castration

この原稿は、実験用マウスにおける前立腺顕微解剖や去勢手術のためのプロトコルについて説明します。また、これらのプロトコルによって生成される代表的な結果を示しています。最後に、我々は長所と、これらのプロトコルの利用を議論します。

マウス前立腺マイクロ解剖と外科的去勢

Mouse models are used extensively to study prostate cancer and other diseases. The mouse is an excellent model with which to study the prostate and has ...

murine-prostate-micro-dissection-and-surgical-castration

Research

JoVE Journal

Medicine

45.2K ビュー.  Johns Hopkins University. この原稿は、実験用マウスにおける前立腺顕微解剖や去勢手術のためのプロトコルについて説明します。また、これらのプロトコルによって生成される代表的な結果を示しています。最後に、我々は長所と、これらのプロトコルの利用を議論します。 

ビデオ: Murine Prostate Micro-dissection and Surgical Castration

Microvascular Decompression: Salient Surgical Principles and Technical Nuances

Trigeminal neuralgia is a disorder associated with severe episodes of lancinating pain in the distribution of the trigeminal nerve. Previous reports indicate that 80-90% of cases are related to compression of the trigeminal nerve by an adjacent vessel. The majority of patients with trigeminal neuralgia eventually require surgical management in order to achieve remission of symptoms. Surgical options for management include ablative procedures (e.g., radiosurgery, percutaneous radiofrequency lesioning, balloon compression, glycerol rhizolysis, etc.) and microvascular decompression. Ablative procedures fail to address the root cause of the disorder and are less effective at preventing recurrence of symptoms over the long term than microvascular decompression. However, microvascular decompression is inherently more invasive than ablative procedures and is associated with increased surgical risks. Previous studies have demonstrated a correlation between surgeon experience and patient outcome in microvascular decompression. In this series of 59 patients operated on by two neurosurgeons (JSN and PEK) since 2006, 93% of patients demonstrated substantial improvement in their trigeminal neuralgia following the procedure&#x2014;with follow-up ranging from 6 weeks to 2 years. Moreover, 41 of 66 patients (approximately 64%) have been entirely pain-free following the operation.
In this publication, video format is utilized to review the microsurgical pathology of this disorder. Steps of the operative procedure are reviewed and salient principles and technical nuances useful in minimizing complications and maximizing efficacy are discussed.

There are many available options for management of the patient with trigeminal neuralgia. Microvascular decompression, while the most invasive of all options, is also the most effective at achieving long term remission of symptoms. Video instruction on how to maximize efficacy and minimize complications with this procedure is described.

微小血管減圧術：顕著な外科手術の原則と技術的なニュアンス

Trigeminal neuralgia is a disorder associated with severe episodes of lancinating pain in the distribution of the trigeminal nerve. Previous reports ...

医学

Evaluation of Nanoparticle Uptake in Tumors in Real Time Using Intravital Imaging

Current technologies for tumor imaging, such as ultrasound, MRI, PET and CT, are unable to yield high-resolution images for the assessment of nanoparticle uptake in tumors at the microscopic level1,2,3, highlighting the utility of a suitable xenograft model in which to perform detailed uptake analyses. Here, we use high-resolution intravital imaging to evaluate nanoparticle uptake in human tumor xenografts in a modified, shell-less chicken embryo model. The chicken embryo model is particularly well-suited for these in vivo analyses because it supports the growth of human tumors, is relatively inexpensive and does not require anesthetization or surgery 4,5. Tumor cells form fully vascularized xenografts within 7 days when implanted into the chorioallantoic membrane (CAM) 6. The resulting tumors are visualized by non-invasive real-time, high-resolution imaging that can be maintained for up to 72 hours with little impact on either the host or tumor systems. Nanoparticles with a wide range of sizes and formulations administered distal to the tumor can be visualized and quantified as they flow through the bloodstream, extravasate from leaky tumor vasculature, and accumulate at the tumor site. We describe here the analysis of nanoparticles derived from Cowpea mosaic virus (CPMV) decorated with near-infrared fluorescent dyes and/or polyethylene glycol polymers (PEG) 7, 8, 9,10,11. Upon intravenous administration, these viral nanoparticles are rapidly internalized by endothelial cells, resulting in global labeling of the vasculature both outside and within the tumor7,12. PEGylation of the viral nanoparticles increases their plasma half-life, extends their time in the circulation, and ultimately enhances their accumulation in tumors via the enhanced permeability and retention (EPR) effect 7, 10,11. The rate and extent of accumulation of nanoparticles in a tumor is measured over time using image analysis software. This technique provides a method to both visualize and quantify nanoparticle dynamics in human tumors.

We present a novel approach to quantify nanoparticle localization in the vasculature of human xenografted tumors using dynamic, real-time intravital imaging in an avian embryo model.

生体内イメージングを使ってリアルタイムで腫瘍におけるナノ粒子の取り込みの評価

Current technologies for tumor imaging, such as ultrasound, MRI, PET and CT, are unable to yield high-resolution images for the assessment of nanoparticle ...

An Orthotopic Murine Model of Human Prostate Cancer Metastasis

Our laboratory has developed a novel orthotopic implantation model of human prostate cancer (PCa). As PCa death is not due to the primary tumor, but rather the formation of distinct metastasis, the ability to effectively model this progression pre-clinically is of high value. In this model, cells are directly implanted into the ventral lobe of the prostate in Balb/c athymic mice, and allowed to progress for 4-6 weeks. At experiment termination, several distinct endpoints can be measured, such as size and molecular characterization of the primary tumor, the presence and quantification of circulating tumor cells in the blood and bone marrow, and formation of metastasis to the lung. In addition to a variety of endpoints, this model provides a picture of a cells ability to invade and escape the primary organ, enter and survive in the circulatory system, and implant and grow in a secondary site. This model has been used effectively to measure metastatic response to both changes in protein expression as well as to response to small molecule therapeutics, in a short turnaround time.

This orthotopic model of human prostate cancer allows for quantification of tumor size, circulating tumor cells, and formation of distinct metastasis to the lung. As cells must escape the primary organ, enter the blood stream, and implant into a secondary site, this model effectively recapitulates the scenario in humans.

ヒト前立腺癌転移の同所マウスモデル

Our laboratory has developed a novel orthotopic implantation model of  human prostate cancer (PCa). As PCa death is not due to the primary tumor, but  ...

Surgical Fixation of Sternal Fractures: Preoperative Planning and a Safe Surgical Technique Using Locked Titanium Plates and Depth Limited Drilling

Different ways to stabilize a sternal fracture are described in literature. Respecting different mechanisms of trauma such as the direct impact to the anterior chest wall or the flexion-compression injury of the trunk, there is a need to retain each sternal fragment in the correct position while neutralizing shearing forces to the sternum. Anterior sternal plating provides the best stability and is therefore increasingly used in most cases. However, many surgeons are reluctant to perform sternal osteosynthesis due to possible complications such as difficulties in preoperative planning, severe injuries to mediastinal organs, or failure of the performed method.
This manuscript describes one possible safe way to stabilize different types of sternal fractures in a step by step guidance for anterior sternal plating using low profile locking titanium plates. Before surgical treatment, a detailed survey of the patient and a three dimensional reconstructed computed tomography is taken out to get detailed information of the fracture&#8217;s morphology. The surgical approach is usually a midline incision. Its position can be described by measuring the distance from upper sternal edge to the fracture and its length can be approximated by the summation of 60 mm for the basis incision, the thickness of presternal soft tissue and the greatest distance between the fragments in case of multiple fractures.
Performing subperiosteal dissection along the sternum while reducing the fracture, using depth limited drilling, and fixing the plates prevents injuries to mediastinal organs and vessels.
Transverse fractures and oblique fractures at the corpus sterni are plated longitudinally, whereas oblique fractures of manubrium, sternocostal separation and any longitudinally fracture needs to be stabilized by a transverse plate from rib to sternum to rib. Usually the high convenience of a patient is seen during follow up as well as a precise reconstruction of the sternal morphology.

Here we present a method to stabilize sternal fractures by using locked titanium plates in a low profile design. Performing subperiosteal dissection along the sternum while reducing the fracture, using depth limited drilling, and fixing the plates provides a safe surgical way.

胸骨骨折の外科的固定：手術計画とロックされたチタン板を使用した安全な手術法と深さリミテッド掘削

Different ways to stabilize a sternal fracture are described in literature. Respecting different mechanisms of trauma such as the direct impact to the ...

A Novel Murine Model of Arteriovenous Fistula Failure: The Surgical Procedure in Detail

The arteriovenous fistula (AVF) still suffers from a high number of failures caused by insufficient remodeling and intimal hyperplasia from which the exact pathophysiology remains unknown. In order to unravel the pathophysiology a murine model of AVF-failure was developed in which the configuration of the anastomosis resembles the preferred situation in the clinical setting. A model was described in which an AVF is created by connecting the venous end of the branch of the external jugular vein to the side of the common carotid artery using interrupted sutures. At a histological level, we observed progressive stenotic intimal lesions in the venous outflow tract that is also seen in failed human AVFs. Although this procedure can be technically challenging due to the small dimensions of the animal, we were able to achieve a surgical success rate of 97% after sufficient training. The key advantage of a murine model is the availability of transgenic animals. In view of the different proposed mechanisms that are responsible for AVF failure, disabling genes that might play a role in vascular remodeling can help us to unravel the complex pathophysiology of AVF failure.

Here we present a murine model of arteriovenous fistula (AVF) failure in which a clinically relevant anastomotic configuration is incorporated. This model can be used to study the pathophysiology and to test possible therapeutic interventions.

動静脈瘻の障害の新規マウスモデル：詳細の外科的手順

The arteriovenous fistula (AVF) still suffers from a high number of failures caused by insufficient remodeling and intimal hyperplasia from which the ...

Pre-clinical Orthotopic Murine Model of Human Prostate Cancer

To study the multifaceted biology of prostate cancer, pre-clinical in vivo models offer a range of options to uncover critical biological information about this disease. The human orthotopic prostate cancer xenograft mouse model provides a useful alternative approach for understanding the specific interactions between genetically and molecularly altered tumor cells, their organ microenvironment, and for evaluation of efficacy of therapeutic regimens. This is a well characterized model designed to study the molecular events of primary tumor development and it recapitulates the early events in the metastatic cascade prior to embolism and entry of tumor cells into the circulation. Thus it allows elucidation of molecular mechanisms underlying the initial phase of metastatic disease. In addition, this model can annotate drug targets of clinical relevance and is a valuable tool to study prostate cancer progression. In this manuscript we describe a detailed procedure to establish a human orthotopic prostate cancer xenograft mouse model.

Prostate cancer is the second most common cause of cancer-related deaths in the United States. An orthotopic cancer model provides a useful approach to understand the biology of prostate cancer and to evaluate the efficacy of therapeutic regimens. This protocol describes detailed steps necessary to establish an orthotopic prostate cancer mouse model.

ヒト前立腺癌の前臨床同所性マウスモデル

To study the multifaceted biology of prostate cancer, pre-clinical in vivo models offer a range of options to uncover critical biological information ...

Intravital Microscopy of Tumor-associated Vasculature Using Advanced Dorsal Skinfold Window Chambers on Transgenic Fluorescent Mice

Tumor and tumor vessel development, as well as tumor response to therapeutics, are highly dynamic biological processes. Histology provides static information and is often not sufficient for a correct interpretation. Intravital evaluation, in which a process is followed in time, provides extra and often unexpected information. With the creation of transgenic animals expressing cell-specific markers and live cell tracers, improvements to imaging equipment, and the development of several imaging chambers, intravital microscopy has become an important tool to better understand biological processes. This paper describes an experimental design for the investigation of tumor vessel development and of therapeutic effects in a spatial and temporal manner. Using this setup, the stage of vessel development, tip cell and lumen formation, blood flow, extravasation, an established vascular bed, and vascular destruction can be visualized and followed. Furthermore, therapeutic effects, intratumoral fate, and the localization of chemotherapeutic compounds can also be followed.

This protocol describes the intravital imaging of transgenic mice expressing cell-specific fluorescent markers. Intravital imaging provides a non-invasive method for high-resolution observations in living animals using implantable windows through which the microscopic visualization of processes is possible. This method is especially useful to study longitudinal processes.

腫瘍関連血管系を使用しての生体顕微鏡が背高度な蛍光トランスジェニック マウスの皮下脂肪のウィンドウの部屋

Tumor and tumor vessel development, as well as tumor response to therapeutics, are highly dynamic biological processes. Histology provides static ...

Using In Vitro Live-cell Imaging to Explore Chemotherapeutics Delivered by Lipid-based Nanoparticles

Conventional imaging techniques can provide detailed information about cellular processes. However, this information is based on static images in an otherwise dynamic system, and successive phases are easily overlooked or misinterpreted. Live-cell imaging and time-lapse microscopy, in which living cells can be followed for hours or even days in a more or less continuous fashion, are therefore very informative. The protocol described here allows for the investigation of the fate of chemotherapeutic nanoparticles after the delivery of doxorubicin (dox) in living cells. Dox is an intercalating agent that must be released from its nanocarrier to become biologically active. In spite of its clinical registration for more than two decades, its uptake, breakdown, and drug release are still not fully understood. This article explores the hypothesis that lipid-based nanoparticles are taken up by the tumor cells and are slowly degraded. Released dox is then translocated to the nucleus. To prevent fixation artifacts, live-cell imaging and time-lapse microscopy, described in this experimental procedure, can be applied.

Using In Vitro Live-cell Imaging to Explore Chemotherapeutics Delivered by Lipid-based Nanoparticles

Live-cell imaging is a powerful tool to visualize dynamic processes. The examination of fixed cells provides only static pictures, which can lead to misinterpretation and confusion about the process. This work presents a method to study uptake, drug release, and intracellular localization of liposomal nanoparticles in living cells.

体外配信生細胞イメージング化学療法を探求する脂質ナノ粒子によってを使用してください。

Conventional imaging techniques can provide detailed information about cellular processes. However, this information is based on static images in an ...

Surgical Treatment for Benign Prostatic Hyperplasia: Holmium Laser Enucleation of the Prostate (HoLEP).

Benign prostatic hyperplasia (BPH) commonly occurs among elderly males. The condition causes symptoms in the lower urinary tract, thereby reducing the quality of life. BPH includes 2 major treatments: medication and surgery. Surgical treatment is often the most effective and final intervention. Holmium laser enucleation of the prostate (HoLEP), one of the most efficient surgical procedures for BPH, is conducted transurethrally. During surgery, the most important and most difficult part is to locate the surgical capsule of the prostate, which is consistent to the idea of anatomic surgery. Different skills may be needed in treating different parts of the prostate. HoLEP tends to be efficient and safe with good hemostatic properties, and able to treat bladder calculus, which may be the complications of BPH. The technique is particularly suitable for prostate of varying volumes and sizes. Indeed, HoLEP also presents certain disadvantages, such as a long learning curve and costly equipment. Regardless, this method may be the "new standard" and the excellent method for the surgical treatment of BPH.

Surgical Treatment for Benign Prostatic Hyperplasia: Holmium Laser Enucleation of the Prostate HoLEP.

Here we present a safe and efficient protocol, holmium laser enucleation of the prostate, to treat benign prostatic hyperplasia.

前立腺肥大症の外科治療: ホルミウム レーザー核出術 (HoLEP) 前立腺の。

Benign prostatic hyperplasia (BPH) commonly occurs among elderly males. The condition causes symptoms in the lower urinary tract, thereby reducing the ...

Micro-dissection of Enamel Organ from Mandibular Incisor of Rats Exposed to Environmental Toxicants

Enamel defects resulting from environmental conditions and ways of life are public health concerns because of their high prevalence. These defects result from altered activity of cells responsible for enamel synthesis named ameloblasts, which present in enamel organ. During amelogenesis, ameloblasts follow a specific and precise sequence of events of proliferation, differentiation, and death. A rat continually growing incisors is a suitable experimental model to study ameloblast activity and differentiation stages in physiological and pathological conditions. Here, we describe a reliable and consistent method to micro-dissect enamel organ of rats exposed to environmental toxicants. The micro-dissected dental epithelia contain secretion- and maturation-stage ameloblasts that may be used for qualitative experiments, such as immunohistochemistry assays and in situ hybridization, as well as for quantitative analyses such as RT-qPCR, RNA-seq, and Western blotting.

Understanding enamel formation and possible alterations requires the study of ameloblast activity. Here, we describe a reliable and consistent method to micro-dissect enamel organs containing secretion- and maturation-stage ameloblasts that may be used for further quantitative and qualitative experimental procedures.

環境有害物質に暴露したラットの下顎からエナメル器官の微細解剖

Enamel defects resulting from environmental conditions and ways of life are public health concerns because of their high prevalence. These defects result ...