We would like to thank the Children&#8217;s Hospital at Montefiore, Department of Pediatrics, for providing support for these studies. We would like to thank the National Alopecia Areata Foundation for providing financial support for conducting studies with C3H/HeJ engrafted mice.

倫理声明：動物に関わるすべての研究は施設のIACUCによって承認されなければならない（以下のデータについては、プロトコルは、モンテフィオーレIACUC、プロトコル＃11-6-240と＃13-7-100によって承認された）。撮影時まだそれらを維持する唯一の目的のために示されているように、動物が軽い麻酔を提供され、このプロトコルに必要ない痛みを伴う手続きはありません。 写真の1.取得<ol><li>印刷されたテキストと紙を用いたゲルイメージャのためのビューのフォーカスとフィールドを設定します。撮影した地域全体の光源の均一性を確認してください。均一性を確保するために、反射写真撮影のための光源を内蔵したゲルイメージャを使用しています。これはさらにグレースケールの分析には適していない動物のシルエットを作成すると同じように、透照光源を使用しないでください。 注：これは関心領域（ROI）との意志を越え、光の平均化を提供する、やや焦点の外にマウスを置きます興味のある非常に小さい（ すなわち、&lt;10ピクセル）の領域について量子的エラーを減らすのに役立ちます。関心の大きな領域は、このマイナーな焦点調節によって影響されず、またそれらは、動物の大きさの違いによって影響される。 </li><li>これは急速な麻酔効果および迅速な回復を提供し、複数の動物を撮影するために最適であるように、：ケタミン（100mg / mlの）/キシラジン（20 mg / ml）で（1 2）を使用して動物を麻酔。 注記：動物が静止画撮影を可能にするのに十分であるときに麻酔が確認された。動物は一般的に10〜15分以内にこの光麻酔から回復したように、獣医師は眼軟膏の使用をお勧めしていません。 </li><li> （できるだけ平行に近い）の垂直配向ゲルイメージャーで麻酔した動物を置きます。 <ol><li>背写真の場合は、延長手足と腹臥位で動物を配置。 </li><li>腹写真のために、動物が横方向に回転しないように注意しながら、仰臥位で動物を配置。 </li&gt; </ol></li><li>撮影した領域でのグレースケール標準を置きます。 </li><li>閉じるアクセスドア。重要：周囲光露光の変化を導入することができる。 </li><li>取得の直線範囲内の関心領域を配置露光Fストップを設定する（Fストップを読み取る）。 注：画像が飽和している場所ほとんどのシステムが表示されます。 </li><li>写真を撮る。 </li><li>変更標準および関心領域の両方を取得（参照の直線範囲内に留まるように、1からFストップを増加または減少させることによって、直線範囲内の関心領域を配置し、別の露出設定にFストップF-停止）。 </li><li>写真を撮る。 </li><li>撮影が完了すると、加温テーブルに動物を配置し、それらが伏臥を維持できるようになるまで監視。ケージに動物を返します。一度、すべての動物が完全に回復している動物施設への動物のグループを返します。 </li></ol>光の吸収2.定量&lt;/ pの&gt; <ol><li>使用した動物の画像上で関心のマーク領域は、ゲルイメージャのためのソフトウェアを提供した。 <ol><li>全動物の背ビューについては、可能な限り横方向に限り延長、下肢への上肢から延びる長方形または楕円形のイメージを使用するように、 図1Aに示すように、箱のいかなる部分も、動物の背中を越えて延びていない。 NOTE：一つは、フリーハンド描画ツールを使用して、関心のある領域をマークできた。 </li><li>骨盤領域をカバーする1と図1（b）に示すように胸のエリアをカバーする1：全体の動物腹ビューについては、2の長方形を使用しています。 </li><li>必要に応じて、関心、 すなわち、薬剤投与の位置、マークのより小さな領域のために。 </li></ol></li><li>グレースケール吸収規格上の関心のマーク領域（複数可）。 </li><li>関心のマークされた領域のそれぞれからレコード吸収。 </li></ol> 3.分析吸収レベルobtaine関心領域の研究開発は、写真間の比較のために、バックグラウンドの標準に正規化する必要があるかもしれない。吸収への曝露の関係は両対数（ すなわち、関係がログ（曝露の間に線形である）とログ（吸収））である。この関係を用いて、関心領域の吸収は、吸収が異なる時点（試験プロトコル内、すなわち、連続測定）で撮影されたものを含む別の写真の間で直接比較することを可能にする標準的なバックグラウンド値に正規化することができる。これらの補正を実行するための手順は、オプションのステップ3.1および3.2に詳述されている。 <ol><li>グレースケールの吸収標準から得られた値を使用して、ログ対ログの（オプション）プロット曲線（露出）（吸収）。 </li><li>次のように（オプション）各写真の標準の変動を調整します。 <ol><li>秒で決定されるように（参照のためおよびFストップ（ステップ1.6で決定されるように）読み取るためFストップを選択1.8）をTEP。 </li><li>読んでFストップは全く写真に標準の平均吸収を計算します。この平均値は参考基準値（RSV）である。 </li><li> （デルタ-ROI-1、デルタ-ROI-2 ... ..delta-ROI-X）規格（デルタ-S）のために、各定義されたROIのためにそれぞれの写真に写っている読書と参照F-ストップ設定間の吸収の差を計算する</li><li> *（デルタROI-X / - F-ストップ+（F-ストップを読んで、標準の吸収RSV）を読んで補正吸収（ROI-X）=吸収（ROI-X）：次のように各ROIに対する補正吸収を計算しますデルタS） </li></ol></li><li>実験データをコンパイルし、連続変数（ すなわち、ANOVA、T検定）のための標準的な統計的手法を用いてデータ解析を行う。 </li></ol>

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Authors have nothing to disclose.

本報告書では詳細な説明は、げっ歯類で脱毛を定量化するための新しい技術の提供されています。この技術は、ほとんどの実験室で容易に利用可能である画像取得と分析のためのゲルイメージャ、機器を利用する。測定は、技術（ 図4から5）における小さな変化に対してロバストであることが示されており、治療モデルにおける視覚脱毛（ 図6）の程度によく相関し?...

脱毛（抜け毛）は複数の原因と心理的、感情的苦痛イベントになることができます。男性型脱毛症は35歳1によって男性の約2/3に影響を及ぼし、脱毛症の最も一般的な原因である。抜け毛の同様のパターンは、多嚢胞性卵巣症候群の女性において観察することができる。これらの疾患の両方において、脱毛はアンドロゲン媒介性である。脱毛症はまた、人口2の1.7％に影響を与える自己免疫疾患、円形脱毛症、として発生する可能性があります。脱毛症は、化学療法3のようないくつかの医学的治療の副作用として発生する可能性があります。化学療法患者の割合が高い（65から85までパーセント）は、脱毛症4,5ある程度の経験する。脱毛の心理的な影響は十分に化学療法の設定で研究されている。化学療法誘発性脱毛症、不安、抑う ​​つ、負の身体イメージをもたらすことができ、自尊心と6,7-幸福の減少感を低下させた。高いPE女性のがん患者のrcentage（百分の47から58）は脱毛は、化学療法の最も外傷側面であると考えて、そして脱毛4,6の恐れのための最大8％の下落治療。脱毛8,9の心理的にも医学的結果を減少させるための治療をサポートするための男性ホルモン性脱毛症における証拠もある。同様に、円形脱毛症は、重度の心理的な影響2、及び脱毛の斑点状の性質が脱毛の他のほとんどの原因はより不快化粧結果につながることが報告されている。 穏やかな抗アンドロゲン作用（ すなわち、スピロノラクトン）を有する薬剤が脱毛症の治療などの限られた成功を収めて使用されていたが、脱毛症のための最初の効果的な薬物療法は、ミノキシジル10だった。この血圧降下は、毛髪の成長を引き起こすの観察された副作用を有しており、現在脱毛症の多くの形態のための局所療法として使用される。しかし、応答はいくつかの科目だけ遅く表示で、しばしば不完全である脱毛ではなく、実際の再成長の10る。フィナステリドは、部分的な全身のアンドロゲン遮断を犠牲にしてアンドロゲン性脱毛症の改善で、その結果、ジヒドロテストステロンのII5α還元酵素をブロック変換を入力する競合的アンタゴニストである。長期的（10年）療法との回答率は50％前後11である。全体として、この分野におけるかなりの研究にもかかわらず、脱毛のための適切な治療は依然として存在しない。 何十年もの間、科学者や臨床医は、臨床試験で頭皮毛髪の成長を測定する方法を検討した。脱毛症を治療する薬剤の開発により、毛髪の成長と、治療に対する特異的応答を測定する、信頼性の高い経済的な低侵襲手段に対する大きな必要性があった。脱毛障害を有する患者における毛髪密度の正確な定量化のための画像解析技術がtechniquの様々な方法を用いて、過去に一貫して有効な結果が得られた定義された頭皮領域14に毛量を定量化するためのデジタル画像12の解析、個々の毛髪及び皮膚病変13、および顕微鏡走査の画像解析を含むエス。 上記の方法論は、臨床試験における発毛促進介入のための有効性の改善された評価を提供している残念ながら、これらの方法は、前臨床研究において、げっ歯類の研究に適用されていない。我々の目的は、研究者は、より正確に脱毛症の種々の形態のための新たな治療アプローチを評価し、比較することを可能にするであろう、マウスにおいて、脱毛を定量化するために一貫して信頼性の高い方法を開発することである。私たちは、茶色や黒い髪を持つマウスにおける毛髪密度の迅速かつ信頼性の高い定量化が可能になりますほとんどの実験室で容易に利用可能な機器を使用して方法論を開発した。この方法はwaxinから化学療法誘発性脱毛症、円形脱毛症、および脱毛症のマウスモデルにおいて試験されているグラム。詳細なプロトコルは、マウスのC57BL / 6及びC3H / HEJ株からの検証データを含む、これらの測定を実行するために提示されている。この技術は、毛幹中での顔料の光吸収を検出することに依存しているように、白マウス又はアルビノマウスにおける発毛を検出するために使用することができない。

<table><tbody><tr><td>KODAK Gel Logic 100 Imaging System</td><td>Eastman Kodak Company, Rochester, NY, USA</td><td></td><td>Gel imager for obtaining and analyzing photographs, must have built in light source for reflective photography.</td></tr><tr><td>The Kodak/Tiffen Q-13 Gray Scale</td><td><a href="http://www.imatest.com/docs/q13/" target="_blank"><img src="/img/new_window.png" /> Imatest</a></td><td></td><td>Greyscale standard</td></tr><tr><td>C57BL/6J Mice</td><td>Jackson Laboratories, Bar Harbor, Maine</td><td></td><td>Mice for representative study</td></tr><tr><td>C3H/HeJ engrafted mouse</td><td>Jackson Laboratories, Bar Harbor, Maine</td><td></td><td>Mice for representative study</td></tr></tbody></table>

a new technique for quantitative analysis of hair loss in mice using grayscale analysis

脱毛症は、男性型脱毛、多嚢胞性卵巣症候群、および円形脱毛症など、さまざまな条件で発生する可能性が脱毛の一般的な形式です。脱毛症は、癌患者における化学療法の副作用として生じ得る。本研究では、私たちの目標は、研究者が正確に評価し、脱毛症のこれらの様々な形態のための新たな治療アプローチを比較することができるようになりますマウスでの脱毛を、定量化するための一貫性と信頼性の高い方法を開発することであった。この方法は、マウスに黒（または灰色）の毛髪の量にラフ比例して発生する光吸収を測定する、マウスの画像を取得し、処理するための標準的なゲルイメージャを利用しています。このようにして定量化されたデータは、その後、標準的な統計的手法（ すなわち、ANOVA、T検定）を用いて分析することができる。この方法は、ワックスの化学療法誘発性脱毛症、円形脱毛症および脱毛症のマウスモデルにおいて試験した。このレポートでは、詳細なプロトコルは、PRESですマウスのC57BL / 6及びC3H / HEJ株からの検証データを含む、これらの測定を行うために取得済み。この新技術は、相対アプリケーションのシンプルさ、ほとんどの研究室で容易に利用可能である機器への依存、そして客観、主観的な評価よりも堅牢である定量的な評価を適用するなど、多くの利点を提供しています。マウスにおける毛髪成長の定量化の改善は、脱毛症モデルの研究を改善し、前臨床試験で有望な新しい治療法の評価を容易にする。

この技術は、C3H / HEJ生着マウス、16円形脱毛症のマウスモデルを用いて検証した。これらの動物は、時間をかけて徐々に進行するグローバル脱毛を開発する。しかし、脱毛パッチで発生し、かなりの変動を導入し、定性的な評価を妨げ、次の1匹のマウスから異なる場合があります。このモデルは、有効性の評価として、マウスの異なる位置での光学密度の測定値の間の相関をテスト?...

Watch this Scientific Journal Video about A New Technique for Quantitative Analysis of Hair Loss in Mice Using Grayscale Analysis at JoVE.com

A New Technique for Quantitative Analysis of Hair Loss in Mice Using Grayscale Analysis

Alopecia is a common form of hair loss which can occur in many different conditions, including as a side-effect of chemotherapy. We have developed a method to quantify hair loss in mice, utilizing a standard gel imager to perform a grayscale analysis, to facilitate study of promising new alopecia therapies.

グレースケール解析を用いたマウスの抜け毛の定量分析のための新たな手法

Alopecia is a common form of hair loss which can occur in many different conditions, including male-pattern hair loss, polycystic ovarian syndrome, and ...

a-new-technique-for-quantitative-analysis-hair-loss-mice-using

Universidad de Cartagena UDEC

Research

JoVE Journal

Biology

8.9K ビュー.  Children's Hospital at Montefiore. Alopecia is a common form of hair loss which can occur in many different conditions, including as a side-effect of chemotherapy. We have developed a method to quantify hair loss in mice, utilizing a standard gel imager to perform a grayscale analysis, to facilitate study of promising new alopecia therapies.

ビデオ: A New Technique for Quantitative Analysis of Hair Loss in Mice Using Grayscale Analysis

Flat Mount Imaging of Mouse Skin and Its Application to the Analysis of Hair Follicle Patterning and Sensory Axon Morphology

Skin is a highly heterogeneous tissue. Intra-dermal structures include hair follicles, arrector pili muscles, epidermal specializations (such as Merkel cell clusters), sebaceous glands, nerves and nerve endings, and capillaries. The spatial arrangement of these structures is tightly controlled on a microscopic scale - as seen, for example, in the orderly arrangement of cell types within a single hair follicle - and on a macroscopic scale - as seen by the nearly identical orientations of thousands of hair follicles within a local region of skin. Visualizing these structures without physically sectioning the skin is possible because of the 2-dimensional geometry of this organ. In this protocol, we show that mouse skin can be dissected, fixed, permeabilized, stained, and clarified as an intact two dimensional object, a flat mount. The protocol allows for easy visualization of skin structures in their entirety through the full thickness of large areas of skin by optical sectioning and reconstruction. Images of these structures can also be integrated with information about position and orientation relative to the body axes.

Mammalian skin contains a diverse array of structures - such as hair follicles and nerve endings - that exhibit distinctive patterns of spatial organization. Analyzing skin as a flat mount takes advantage of the 2-dimensional geometry of this tissue to produce full-thickness high-resolution images of skin structures.

マウス皮膚や毛包パターニングおよび感覚軸索形態の解析への応用のフラットマウントイメージング

Skin is a highly heterogeneous tissue. Intra-dermal structures include hair follicles, arrector pili muscles, epidermal specializations (such as Merkel ...

神経科学

Combined Recording of Mechanically Stimulated Afferent Output and Nerve Terminal Labelling in Mouse Hair Follicle Lanceolate Endings

A novel dissection and recording technique is described for monitoring afferent firing evoked by mechanical displacement of hairs in the mouse pinna. The technique is very cost-effective and easily undertaken with materials commonly found in most electrophysiology laboratories, or easily purchased. The dissection is simple and fast, with the mechanical displacement provided by a generic electroceramic wafer controlled by proprietary software. The same software also records and analyses the electroneurogram output. The recording of the evoked nerve activity is through a commercial differential amplifier connected to fire-polished standard glass microelectrodes. Helpful tips are given for improving the quality of the preparation, the stimulation and the recording conditions to optimize recording quality. The system is suitable for assaying the electrophysiological and optical properties of lanceolate terminals of palisade endings of hair follicles, as well as the outcomes from their pharmacological and/or genetic manipulation. An example of combining electrical recording with mechanical stimulation and labeling with a styryl pyridinium vital dye is given.

A simple and novel technique for recording afferent discharge due to mechanical stimulation of lanceolate terminals of palisade endings innervating mouse ear skin hair follicles is presented.

マウスの毛包披針形エンディングで機械的に刺激された求心性の出力および神経ターミナルラベリングの複合記録

A novel dissection and recording technique is described for monitoring afferent firing evoked by mechanical displacement of hairs in the mouse pinna. The ...

Optical Monitoring of Living Nerve Terminal Labeling in Hair Follicle Lanceolate Endings of the Ex Vivo Mouse Ear Skin

A novel dissection and recording technique is described for optical monitoring staining and de-staining of lanceolate terminals surrounding hair follicles in the skin of the mouse pinna. The preparation is simple and relatively fast, reliably yielding extensive regions of multiple labeled units of living nerve terminals to study uptake and release of styryl pyridinium dyes extensively used in studies of vesicle recycling. Subdividing the preparations before labeling allows test vs. control comparisons in the same ear from a single individual. Helpful tips are given for improving the quality of the preparation, the labeling and the imaging parameters. This new system is suitable for assaying pharmacologically and mechanically-induced uptake and release of these vital dyes in lanceolate terminals in both wild-type and genetically modified animals. Examples of modulatory influences on labeling intensity are given.

Optical Monitoring of Living Nerve Terminal Labeling in Hair Follicle Lanceolate Endings of the Ex Vivo Mouse Ear Skin

Here we describe a novel preparation for imaging live lanceolate sensory terminals of palisade endings that innervate mouse ear skin hair follicles during staining and destaining with styryl pyridinium dyes.

の毛包披針形語尾に神経末端標識リビングの光監視<em&gt; ex vivoで</em&gt;マウスの耳皮膚

A novel dissection and recording technique is described for optical monitoring staining and de-staining of lanceolate terminals surrounding hair follicles ...

Bioluminescence-Based Tumor Quantification Method for Monitoring Tumor Progression and Treatment Effects in Mouse Lymphoma Models

Although bioluminescence imaging (BLI) shows promise for monitoring tumor burden in animal models of cancer, these analyses remain mostly qualitative. Here we describe a method for bioluminescence imaging to obtain a semi-quantitative analysis of tumor burden and treatment response. This method is based on the calculation of a luminoscore, a value that allows comparisons of two animals from the same or different experiments. Current BLI instruments enable the calculation of this luminoscore, which relies mainly on the acquisition conditions (back and front acquisitions) and the drawing of the region of interest (manual markup around the mouse). Using two previously described mouse lymphoma models based on cell engraftment, we show that the luminoscore method can serve as a noninvasive way to verify successful tumor cell inoculation, monitor tumor burden, and evaluate the effects of in situ cancer treatment (CpG-DNA). Finally, we show that this method suits different experimental designs. We suggest that this method be used for early estimates of treatment response in preclinical small-animal studies.

Bioluminescence imaging is a well-known tool for localizing tumors and metastases, but quantification of these images often requires complex calculations and particular instruments. We describe the easy-to-use luminoscore method, based on precise acquisition conditions, requiring no calculations, and enabling tumor burden and treatment response to be monitored in mouse models.

マウスリンパ腫モデルで腫瘍の進行と治療効果を監視するための生物発光に基づく腫瘍の定量法

Although bioluminescence imaging (BLI) shows promise for monitoring tumor burden in animal models of cancer, these analyses remain mostly qualitative. ...

医学

Analysis of Craniomaxillofacial Malformations in Mice Using Three-dimensional Microcomputed Tomography

To model craniofacial malformations caused by vitamin A deficiency (VAD), we expressed a dominant-negative retinoid receptor mutation in osteoblasts to specifically inhibit RAR transcriptional activity in mice. This approach allowed us to investigate the effects of VAD on cranial hypomineralization, mandibular deformity, and clavicular hypoplasia in clinical cases. In this study, microcomputed tomography (microCT) scanning of the craniomaxillofacial region of mice represented a valuable tool for studying the growth and development of this animal model. The manual estimation of images is both time-consuming and inaccurate. Hence, here, we present a straightforward, efficient, and accurate approach for segmenting and quantifying the microCT images of each craniomaxillofacial bone. MicroCT software was used to slice the mandible, frontal bone, parietal bone, nasal bone, premaxilla, maxilla, interparietal bone, and occipital bone of mice and measure their corresponding lengths and widths. This segmentation method can be applied to study growth and development in developmental biology, biomedicine, and other related sciences and allows researchers to analyze the effects of genetic mutations on individual craniofacial bones.

Here, we describe in depth a method, which is based on microcomputed tomography, to segment and measure 3D models of craniomaxillofacial bones in mice, for better assessment of craniomaxillofacial bone development in mice than what is possible with current methods.

3次元マイクロコンピュータ断層撮影法によるマウス頭蓋顎顔面奇形の解析

To model craniofacial malformations caused by vitamin A deficiency (VAD), we expressed a dominant-negative retinoid receptor mutation in osteoblasts to ...

発生生物学

Rapid Genetic Analysis of Epithelial-Mesenchymal Signaling During Hair Regeneration

Hair follicle morphogenesis, a complex process requiring interaction between epithelia-derived keratinocytes and the underlying mesenchyme, is an attractive model system to study organ development and tissue-specific signaling. Although hair follicle development is genetically tractable, fast and reproducible analysis of factors essential for this process remains a challenge. Here we describe a procedure to generate targeted overexpression or shRNA-mediated knockdown of factors using lentivirus in a tissue-specific manner. Using a modified version of a hair regeneration model 5, 6, 11, we can achieve robust gain- or loss-of-function analysis in primary mouse keratinocytes or dermal cells to facilitate study of epithelial-mesenchymal signaling pathways that lead to hair follicle morphogenesis. We describe how to isolate fresh primary mouse keratinocytes and dermal cells, which contain dermal papilla cells and their precursors, deliver lentivirus containing either shRNA or cDNA to one of the cell populations, and combine the cells to generate fully formed hair follicles on the backs of nude mice. This approach allows analysis of tissue-specific factors required to generate hair follicles within three weeks and provides a fast and convenient companion to existing genetic models.

Tissue-specific analysis of a hair follicle regeneration model using lentivirus to mediate gain- or loss-of-function.

髪の再生の間に上皮間葉シグナル伝達の迅速遺伝子解析

Hair follicle morphogenesis, a complex process requiring interaction between epithelia-derived keratinocytes and the underlying mesenchyme, is an ...

生物学

Digital Planimetry for Assessing Wound Closure Kinetics in a Mouse Model

Chronic wounds, due to their high prevalence, are a serious global health concern. Effective therapeutic strategies can significantly accelerate healing, thereby reducing the risk of complications and alleviating the economic burden on healthcare systems. Although numerous experimental studies have investigated wound healing, most rely on qualitative observations or quantitative direct measurements. The objective of this study was to standardize an indirect wound measurement method using digital planimetry, incorporating digital scaling and segmentation. This approach addresses the lack of detailed, step-by-step methodologies for accurate wound assessment. A photodocumentation booth was designed and constructed, and computer-assisted digital planimetry tools were employed to minimize variability in measurements of the wound area, perimeter, and the distance from the wound center to its edges. A circular traumatic wound (5 mm in diameter) was created on the dorsal midline at the shoulder blade level of male CD1 mice (n = 4, 10 weeks old, 30-35 g). Wound evolution was photodocumented for 14 days using the custom-designed photo booth, which controlled lighting conditions, focal distance, and subject positioning. Scaling and wound measurements were performed using segmentation in ImageJ software, and statistical analysis was conducted using statistical analysis software. The kinetics of wound closure showed a slight increase in wound size and perimeter between day 0 and day 2, followed by a gradual decrease until complete closure by day 14. The photodocumentation booth and computer-assisted digital planimetry enabled quantitative measurements with minimal variability. In conclusion, these tools provide a reliable and reproducible method for evaluating wound closure kinetics in pre-clinical models.

Wounds represent a global health challenge. This study developed a standardized photo booth utilizing digital planimetry to minimize wound measurement variability. Monitoring wounds in mice over 14 days revealed an initial increase in wound area and perimeter, followed by gradual closure. This methodology may aid in evaluating wound closure kinetics in pre-clinical models.

マウスモデルにおける創傷閉鎖速度を評価するためのデジタルプラニメトリー

Chronic wounds, due to their high prevalence, are a serious global health concern. Effective therapeutic strategies can significantly accelerate healing, ...

生物工学

Non-invasive Skeletal Muscle Quantification in Small Animals Using Micro-computed Tomography

Skeletal muscle size, mass, and composition are critical properties for studying metabolic and muscle-related diseases, as they directly impact the understanding of disease progression and treatment outcomes. Quantifying a live animal's lean, adipose, and skeletal mass is important in metabolic, physiology, pharmacologic, and geroscience studies. However, obtaining accurate body composition measurements, especially of lean mass, remains challenging due to the inherent limitations of conventional assessment techniques. Micro-computed tomography (micro-CT) is a non-invasive radiological technique that enables high-resolution visualization of internal structures in small animal models. A standardized micro-CT method can significantly enhance translational research with more reliable and impactful results, particularly during aging studies or at different time points within the same animal. Despite its potential, the lack of standardization in image acquisition and analysis methods significantly hinders the comparability of results across different studies. Herein, we present a comprehensive and detailed low-cost protocol for lean mass analysis using micro-CT to address these challenges and promote consistency in research involving small animal models.

This protocol uses micro-computed tomography to enable the cost-effective quantification of lean mass, including skeletal muscle and visceral tissue, adipose tissue, and skeletal tissue in small animals. It distinguishes between lean and adipose tissue, which offers significant advantages for biomedical research, particularly in translational research in small animals.

マイクロコンピュータ断層撮影法を用いた小動物における非侵襲的骨格筋定量化

Skeletal muscle size, mass, and composition are critical properties for studying metabolic and muscle-related diseases, as they directly impact the ...

Non-Invasive Visualization of Nailbed Microvascular Morphology in Mice Using Capillaroscopy

Imaging microcapillary networks of the skin in humans using nailfold capillaroscopy (NFC) has underscored the importance of microcirculation as a target organ system in critical systemic illnesses. Nailfold capillaroscopy is applied clinically to detect peripheral microvascular dysfunction and abnormalities in a range of systemic conditions, including rheumatic, cardiac, ocular (e.g., glaucoma), and endocrine disorders (e.g., hypertension and diabetes mellitus). NFC is useful not only in detecting peripheral systemic microvasculature disruption but also in assessing drug efficacy. However, translating clinical NFC findings to animal disease models can be challenging. Detecting microvascular dysfunction or abnormalities in animals is often invasive (e.g., endoscopic), carried out ex vivo (e.g., post-mortem imaging of tissues), or expensive, requiring specialized equipment such as those used in microcomputed tomography and photoacoustic imaging techniques. Developing quick, non-invasive, and inexpensive techniques to image peripheral microvasculature in animal models of disease is warranted to decrease research expenses and increase translatability to the clinic.
Capillaroscopy has previously been used to visualize the nailfold microvasculature in animal models, including in guinea pigs and mice, thus demonstrating the capability of capillaroscopy as a non-invasive imaging tool in animal models. This study provides a protocol that applies capillaroscopy to a mouse nailbed, allowing researchers to easily and inexpensively assess the morphology of its microvasculature. Representative images of typical nailbed microvascular architecture in wild-type mice using two commonly used laboratory strains, SV129/S6 and C57/B6J, are provided. Further studies using this method are essential for applying nailbed capillaroscopy to a wide range of mouse disease models with peripheral microvascular abnormalities.

Capillaroscopy is an accessible tool for direct, inexpensive, and non-invasive visualization of microvasculature. The goal of this protocol is to enable researchers to use capillaroscopy for the visualization of peripheral microvascular morphology in the nailbeds of mice.

キャピラスコープ法を用いたマウスの爪床微小血管形態の非侵襲的可視化

Imaging microcapillary networks of the skin in humans using nailfold capillaroscopy (NFC) has underscored the importance of microcirculation as a target ...

A Technique for Human Skin Grafting in a Mouse Model

Source: Moss, M. I. et al., <a href="https://appjove.unicartagenaproxy.elogim.com/v/64040">Xenograft Skin Model to Manipulate Human Immune Responses&#160;In Vivo.</a>&#160;J. Vis. Exp. (2022)
This video demonstrates a method for grafting human skin onto an immunodeficient mouse with a suppressed neutrophil response. The method involves applying human split-thickness skin to the mouse&#39;s graft bed, sealing it, and covering it for recovery. Eventually, the human skin adheres to the mouse&#39;s skin, establishing vascularization and circulation, resulting in a successful skin xenograft.

マウスモデルにおけるヒト皮膚移植技術

All procedures involving animal models have been reviewed by the local institutional animal care committee and the JoVE veterinary review board.
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