この研究は、ヘルスリサーチおよびゲノムカナダのカナダの協会によってサポートされていました。 JSは、医学研究、ハートとカナダのストローク財団とカナダ糖尿病協会のためのアルバータヘリテージ財団からの給与支援賞を保持しています。研究室では、サポニン、透過性繊維の技術の取得中にOroborosインスツルメンツの技術支援に感謝したい。

<ol>
	<li>Saks, V. A., Veksler, V. I., Kuznetsov, A. V. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Permeabilized+cell+and+skinned+fiber+techniques+in+studies+of+mitochondrial+function+in+vivo.">Permeabilized cell and skinned fiber techniques in studies of mitochondrial function in vivo.</a> Mol Cell Biochem. 184, 81-100 (1998).</li><li>Gnaiger, E., Kuznetsov, A. V., Schneeberger, S., Heldmaier, G., Klingenspor, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Mitochondria+in+the+cold.">Mitochondria in the cold.</a> Life in the Cold. , 431-442 (2000).</li><li>Rasmussen, H. N., Rasmussen, U. F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Oxygen+solubilities+of+media+used+in+electrochemical+respiration+measurements.">Oxygen solubilities of media used in electrochemical respiration measurements.</a> Anal Biochem. 319, 105-113 (2003).</li><li>Visscher, G. D. e., Rooker, S., Jorens, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Pentobarbital+fails+to+reduce+cerebral+oxygen+consumption+early+after+non-hemorrhagic+closed+head+injury+in+rats.">Pentobarbital fails to reduce cerebral oxygen consumption early after non-hemorrhagic closed head injury in rats.</a> J Neurotrauma. 22, 793-806 (2005).</li><li>Kuznetsov, A. V., Veksler, V., Gellerich, F. N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Analysis+of+mitochondrial+function+in+situ+in+permeabilized+muscle+fibers,+tissues+and+cells.">Analysis of mitochondrial function in situ in permeabilized muscle fibers, tissues and cells.</a> Nat Protoc. 3, 965-976 (2008).</li><li>Gnaiger, E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Oxygen+conformance+of+cellular+respiration.+A+perspective+of+mitochondrial+physiology.">Oxygen conformance of cellular respiration. A perspective of mitochondrial physiology.</a> Adv Exp Med Biol. 543, 39-55 (2003).</li><li>Gnaiger, E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Capacity+of+oxidative+phosphorylation+in+human+skeletal+muscle:+new+perspectives+of+mitochondrial+physiology.">Capacity of oxidative phosphorylation in human skeletal muscle: new perspectives of mitochondrial physiology.</a> Int J Biochem Cell Biol. 41, 1837-1845 .</li><li>Sena, S., Hu, P., Zhang, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Impaired+insulin+signaling+accelerates+cardiac+mitochondrial+dysfunction+after+myocardial+infarction.">Impaired insulin signaling accelerates cardiac mitochondrial dysfunction after myocardial infarction.</a> J Mol Cell Cardiol. 46, 910-918 (2009).</li><li>Boudina, S., Sena, S., O'Neill, B. T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Reduced+mitochondrial+oxidative+capacity+and+increased+mitochondrial+uncoupling+impair+myocardial+energetics+in+obesity.">Reduced mitochondrial oxidative capacity and increased mitochondrial uncoupling impair myocardial energetics in obesity.</a> Circulation. 112, 2686-2695 (2005).</li><li>Lenaz, G., Genova, M. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Structure+and+organization+of+mitochondrial+respiratory+complexes:+a+new+understanding+of+an+old+subject.">Structure and organization of mitochondrial respiratory complexes: a new understanding of an old subject.</a> Antioxid Redox Signal. 12, 961-1008 .</li><li>Lemieux, H., Hoppel, C. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Mitochondria+in+the+human+heart.">Mitochondria in the human heart.</a> J Bioenerg Biomembr. 41, 99-106 (2009).</li><li>O, . <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Retarded+diffusion+of+ADP+in+cardiomyocytes:+possible+role+of+mitochondrial+outer+membrane+and+creatine+kinase+in+cellular+regulation+of+oxidative+phosphorylation.">Retarded diffusion of ADP in cardiomyocytes: possible role of mitochondrial outer membrane and creatine kinase in cellular regulation of oxidative phosphorylation.</a> Biochim Biophys Acta. 1144, 134-148 (1993).</li><li>Endo, M., Kitazawa, T., Morad, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=E-C+coupling+studies+in+skinned+cardiac+fibers.">E-C coupling studies in skinned cardiac fibers.</a> Biophysical Aspects of Cardiac Muscle. , 307-327 (1978).</li><li>Veksler, V. I., Kuznetsov, A. V., Sharov, V. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Mitochondrial+respiratory+parameters+in+cardiac+tissue:+a+novel+method+of+assessment+by+using+saponin-skinned+fibers.">Mitochondrial respiratory parameters in cardiac tissue: a novel method of assessment by using saponin-skinned fibers.</a> Biochim Biophys Acta. 892, 191-196 (1987).</li><li>Bangham, A. D., Horne, R. W., Glauert, A. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Action+of+saponin+on+biological+cell+membranes.">Action of saponin on biological cell membranes.</a> Nature. , 196-952 (1962).</li><li>Daum, G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Lipids+of+mitochondria.">Lipids of mitochondria.</a> Biochim Biophys Acta. 822, 1-42 (1985).</li><li>Milner, D. J., Mavroidis, M., Weisleder, N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Desmin+cytoskeleton+linked+to+muscle+mitochondrial+distribution+and+respiratory+function.">Desmin cytoskeleton linked to muscle mitochondrial distribution and respiratory function.</a> J Cell Biol. 150, 1283-1298 (2000).</li><li>Skladal, D., Sperl, W., Schranzhofer, R., Skladal, E., Gellerich, F., Wyss, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Preservation+of+mitochondrial+functions+in+human+skeletal+muscle+during+storage+in+high+energy+preservation+solution+(HEPS).">Preservation of mitochondrial functions in human skeletal muscle during storage in high energy preservation solution (HEPS).</a> What is Controlling Life?. , 268-271 (1994).</li><li>Kuznetsov, A. V., Wiedemann, F. R., Winkler, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Use+of+saponin-permeabilized+muscle+fibers+for+the+diagnosis+of+mitochondrial+diseases.">Use of saponin-permeabilized muscle fibers for the diagnosis of mitochondrial diseases.</a> Biofactors. 7, 221-223 (1998).</li><li>Gnaiger, E., Dykens, J., Will, Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Polarographic+oxygen+sensors,+the+oxygraph+and+high-resolution+respirometry+to+assess+mitochondrial+function.">Polarographic oxygen sensors, the oxygraph and high-resolution respirometry to assess mitochondrial function.</a> Drug-Induced Mitochondrial Dysfunction. , 327-352 (2008).</li></ol>

利害の衝突は宣言されません。

サポニン透過性心臓繊維の技術は、in vitroおよびミトコンドリアOXPHOSの酸素消費量の in vivo評価の間のユニークな妥協を提供しています。この手法の利点は、細胞構造が維持されるとして単離ミトコンドリアに比べて増加した生理学的関連性などがあります。原形質膜が劣化している間、ミトコンドリア12を含む細胞内膜構造、14、筋小胞体14は 、?...

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		<div>
		<table border="1">
		<thead>
		<tr>
		<th>Material Name</th>
		<th>Type</th>
		<th>Company</th>
		<th>Catalogue Number</th>
		<th>Comment</th>
		</tr>
		</thead>
		<tbody>
		
<tr>
<td>100% Ethanol</td>
<td>&nbsp;</td>
<td>Fisher Scientific</td>
<td>HC600</td>
<td>&nbsp;</td>
<tr>
<td>70% Ethanol</td>
<td>&nbsp;</td>
<td>Fisher Scientific</td>
<td>HC-1000</td>
<td>&nbsp;</td>
<tr>
<td>Adenosine 5&#x2032;-diphosphate monopotassium salt dihydrate (ADP)</td>
<td>&nbsp;</td>
<td>Sigma</td>
<td>A5285</td>
<td>&nbsp;</td>
<tr>
<td>Albumin from bovine serum essentially fatty acid&#x2013;free</td>
<td>&nbsp;</td>
<td>Sigma</td>
<td>A-6003</td>
<td>&nbsp;</td>
<tr>
<td>Antimycin A</td>
<td>&nbsp;</td>
<td>Sigma</td>
<td>A8674</td>
<td>&nbsp;</td>
<tr>
<td>Ascobic acid</td>
<td>&nbsp;</td>
<td>Sigma</td>
<td>A4403</td>
<td>&nbsp;</td>
<tr>
<td>Adenosine 5&#x2032;-triphosphate disodium salt hydrate (ATP)</td>
<td>&nbsp;</td>
<td>Sigma</td>
<td>A2383</td>
<td>&nbsp;</td>
<tr>
<td>Atractyloside</td>
<td>&nbsp;</td>
<td>Sigma</td>
<td>A6882</td>
<td>&nbsp;</td>
<tr>
<td>Calcium carbonate</td>
<td>&nbsp;</td>
<td>Sigma</td>
<td>C4830</td>
<td>&nbsp;</td>
<tr>
<td>Carbonyl cyanide p-(trifluoromethoxy) phenylhydrazone (FCCP)</td>
<td>&nbsp;</td>
<td>Sigma</td>
<td>C2920</td>
<td>&nbsp;</td>
<tr>
<td>Cytochrome c</td>
<td>&nbsp;</td>
<td>Sigma</td>
<td>C7752</td>
<td>&nbsp;</td>
<tr>
<td>Digitonin</td>
<td>&nbsp;</td>
<td>Sigma</td>
<td>D141</td>
<td>&nbsp;</td>
<tr>
<td>Dithiothreitol</td>
<td>&nbsp;</td>
<td>Sigma</td>
<td>D9779</td>
<td>&nbsp;</td>
<tr>
<td>Ethylene glycol-bis-(2-aminoethylether)-N,N,N&#x2032;,N&#x2032;-tetraacetic acid (EGTA)</td>
<td>&nbsp;</td>
<td>Sigma</td>
<td>E4378</td>
<td>&nbsp;</td>
<tr>
<td>Glutamic acid</td>
<td>&nbsp;</td>
<td>Sigma</td>
<td>27647</td>
<td>&nbsp;</td>
<tr>
<td>HEPES</td>
<td>&nbsp;</td>
<td>Sigma</td>
<td>H4034</td>
<td>&nbsp;</td>
<tr>
<td>Imidazole</td>
<td>&nbsp;</td>
<td>Sigma</td>
<td>I5513</td>
<td>&nbsp;</td>
<tr>
<td>Ketamine</td>
<td>&nbsp;</td>
<td>Pfizer</td>
<td>&nbsp;</td>
<td>Ketaset</td>
</tr>
<tr>
<td>Lactobionic acid</td>
<td>&nbsp;</td>
<td>Sigma</td>
<td>153516</td>
<td>&nbsp;</td>
<tr>
<td>Magnesium chloride (MgCl2)</td>
<td>&nbsp;</td>
<td>Sigma</td>
<td>M9272</td>
<td>&nbsp;</td>
<tr>
<td>Magnesium chloride hexahydrate (MgCl2&#x2219;6H2O)</td>
<td>&nbsp;</td>
<td>Sigma</td>
<td>M9272</td>
<td>&nbsp;</td>
<tr>
<td>Malic acid</td>
<td>&nbsp;</td>
<td>Sigma</td>
<td>M1000</td>
<td>&nbsp;</td>
<tr>
<td>MES</td>
<td>&nbsp;</td>
<td>Sigma</td>
<td>M3671</td>
<td>&nbsp;</td>
<tr>
<td>N,N,N&#x2019;,N&#x2019;-Tetramethyl- pphenylenediamine Dihydrochloride (TMPD)</td>
<td>&nbsp;</td>
<td>Sigma</td>
<td>T3134</td>
<td>&nbsp;</td>
<tr>
<td>Oligomycin</td>
<td>&nbsp;</td>
<td>Sigma</td>
<td>O4876</td>
<td>&nbsp;</td>
<tr>
<td>Phosphocreatine</td>
<td>&nbsp;</td>
<td>Sigma</td>
<td>P7936</td>
<td>&nbsp;</td>
<tr>
<td>Potassium Chloride</td>
<td>&nbsp;</td>
<td>Sigma</td>
<td>P9541</td>
<td>&nbsp;</td>
<tr>
<td>Potassium Hydroxide</td>
<td>&nbsp;</td>
<td>Sigma</td>
<td>P5958</td>
<td>&nbsp;</td>
<tr>
<td>Potassium cyanide</td>
<td>&nbsp;</td>
<td>Fluka</td>
<td>60178</td>
<td>&nbsp;</td>
<tr>
<td>Potassium phosphate monobasic</td>
<td>&nbsp;</td>
<td>Sigma</td>
<td>P5655</td>
<td>&nbsp;</td>
<tr>
<td>Rotenone</td>
<td>&nbsp;</td>
<td>Sigma</td>
<td>R8875</td>
<td>&nbsp;</td>
<tr>
<td>Saponin</td>
<td>&nbsp;</td>
<td>Sigma</td>
<td>47036</td>
<td>&nbsp;</td>
<tr>
<td>Sodium Pentobarbital</td>
<td>&nbsp;</td>
<td>Ceva Sante Animale</td>
<td>1715 138</td>
<td>Conc. 54.7 mg/ml</td>
</tr>
<tr>
<td>Sodium pyruvate</td>
<td>&nbsp;</td>
<td>Sigma</td>
<td>P2256</td>
<td>&nbsp;</td>
<tr>
<td>Succinic acid</td>
<td>&nbsp;</td>
<td>Sigma</td>
<td>S3674</td>
<td>&nbsp;</td>
<tr>
<td>Sucrose</td>
<td>&nbsp;</td>
<td>Sigma</td>
<td>S7903</td>
<td>&nbsp;</td>
<tr>
<td>Taurine</td>
<td>&nbsp;</td>
<td>Sigma</td>
<td>T8691</td>
<td>&nbsp;</td>
<tr>
<td>Xylazine</td>
<td>&nbsp;</td>
<td>Bayer</td>
<td>&nbsp;</td>
<td>Rompun</td>
</tr>
<tr>
<td>ddH2O</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<tr>
<td>Ice</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<tr>
<td>Oroboros Oxygraph-2k</td>
<td>&nbsp;</td>
<td>Oroboros Instruments</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<tr>
<td>Kimwipes</td>
<td>&nbsp;</td>
<td>VWR</td>
<td>21905-026</td>
<td>&nbsp;</td>
<tr>
<td>15ml polypropylene centrifuge tubes</td>
<td>&nbsp;</td>
<td>VWR</td>
<td>89004-368</td>
<td>&nbsp;</td>
<tr>
<td>50ml polypropylene centrifuge tubes</td>
<td>&nbsp;</td>
<td>VWR</td>
<td>89004-364</td>
<td>&nbsp;</td>
<tr>
<td>Straight Jewelers Forceps</td>
<td>&nbsp;</td>
<td>George Tiemann &amp; Co.</td>
<td>160-50B</td>
<td>&nbsp;</td>
<tr>
<td>Curved Jewelers Forceps</td>
<td>&nbsp;</td>
<td>George Tiemann &amp; Co.</td>
<td>160-57B</td>
<td>&nbsp;</td>
<tr>
<td>Straight Surgery Scissors</td>
<td>&nbsp;</td>
<td>George Tiemann &amp; Co.</td>
<td>105-402</td>
<td>&nbsp;</td>
<tr>
<td>Sterile Surgical Blade</td>
<td>&nbsp;</td>
<td>VWR</td>
<td>BD371610</td>
<td>&nbsp;</td>
<tr>
<td>0.45-&mu;m Syringe filters</td>
<td>&nbsp;</td>
<td>VWR</td>
<td>CA28145-485</td>
<td>&nbsp;</td>
<tr>
<td>pH meter</td>
<td>&nbsp;</td>
<td>VWR</td>
<td>CA11388-308</td>
<td>&nbsp;</td>
<tr>
<td>Glass Petri dishes</td>
<td>&nbsp;</td>
<td>VWR</td>
<td>89000-300</td>
<td>&nbsp;</td>
<tr>
<td>12-well Polystyrene Tissue Culture Plates</td>
<td>&nbsp;</td>
<td>VWR</td>
<td>82050-926</td>
<td>&nbsp;</td>
<tr>
<td>Plate Stirrer</td>
<td>&nbsp;</td>
<td>VWR</td>
<td>97042-594</td>
<td>&nbsp;</td>
<tr>
<td>Fisherbrand Microbars</td>
<td>&nbsp;</td>
<td>Fisher Scientific</td>
<td>14-511-67</td>
<td>&nbsp;</td>
<tr>
<td>Weigh Scale</td>
<td>&nbsp;</td>
<td>VWR</td>
<td>CA11278-162</td>
<td>&nbsp;</td>
<tr>
<td>10&mu;l Hamilton Micro Syringe</td>
<td>&nbsp;</td>
<td>Fisher Scientific</td>
<td>14-815-1</td>
<td>&nbsp;</td>
<tr>
<td>25&#x03BC;l Hamilton Micro Syringe</td>
<td>&nbsp;</td>
<td>Fisher Scientific</td>
<td>14-824-7</td>
<td>&nbsp;</td>
<tr>
<td>50&mu;l Hamilton Micro Syringe</td>
<td>&nbsp;</td>
<td>Fisher Scientific</td>
<td>14-824-5</td>
<td>&nbsp;</td>
<tr>
<td>Nalgene Squeeze Bottles</td>
<td>&nbsp;</td>
<td>Wilkem Scientific</td>
<td>LNA2407-1000</td>
<td>&nbsp;</td>
<tr>
<td>Polystyrene Weighing Dishes</td>
<td>&nbsp;</td>
<td>VWR</td>
<td>89106-750</td>
<td>&nbsp;</td>
<tr>
<td>Dissecting Microscope</td>
<td>&nbsp;</td>
<td>Olympus</td>
<td>&nbsp;</td>
<td>&nbsp;</td>		</tbody>
		</table>
		</div>

respirometric oxidative phosphorylation assessment in saponin-permeabilized cardiac fibers

ミトコンドリアはエネルギー代謝、酸化ストレス、アポトーシス、老化、ミトコンドリアencephalomyopathiesおよび薬物毒性に関与しているとして、ミトコンドリアの機能の調査は、心臓生理学の重要なパラメータを表します。これを考えると、心臓のミトコンドリアの機能を測定する技術が求められています。ミトコンドリアの機能を測定するための統合的アプローチを採用して一つの技術は呼吸計測の酸化的リン酸化（OXPHOS）分析です。 呼吸計測OXPHOSアセスメントの原則は、クラーク電極を利用して酸素濃度を測定することを中心としています。透過処理繊維束は、酸素、密閉チャンバーの低下で酸素濃度を消費するように。選択された基質阻害剤 - 脱共役剤の滴定のプロトコルを使用して、電子は、ミトコンドリアの機能の評価を可能にする、電子伝達鎖の特定部位に用意されています。ミトコンドリアの機能の呼吸計測分析に先立ち、機械的および化学的準備のテクニックは、筋線維の筋鞘を透過するために利用されています。化学物質の透過性は、細胞構造を維持しながら選択的に細胞膜に穴をサポニン採用しています。 本論文では、徹底的にミトコンドリアOXPHOSを評価するために酸素消費量測定用のサポニン肌の心臓の繊維を準備するために必要な手順を説明します。また、アドバイスだけでなく、特定の基質のトラブルシューティングを、電子輸送チェーンの特定の部位でミトコンドリアの機能を決定するために使用されることがあります阻害剤とuncouplersが提供されています。重要なのは、記述されているプロトコルは、簡単に様々な動物モデルおよびヒト試料の心臓および骨格組織に適用されることがあります。

Watch this Scientific Journal Video about Respirometric Oxidative Phosphorylation Assessment in Saponin-permeabilized Cardiac Fibers at JoVE.com

Respirometric Oxidative Phosphorylation Assessment in Saponin-permeabilized Cardiac Fibers

呼吸計測の酸化的リン酸化の解析と組み合わせてサポニン透過処理繊維の調製物は、ミトコンドリアの機能の統合的な評価を提供します。生理的および病的状態におけるミトコンドリア呼吸はミトコンドリアの相互作用、形態および生化学を含む様​​々な規制の影響を反映することができます。

サポニン透過性心臓繊維の呼吸計測の酸化的リン酸化の評価

Investigation of mitochondrial function represents an important parameter of cardiac physiology as mitochondria are involved in energy metabolism, ...

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JoVE Journal

Biology

24.5K ビュー.  University of Calgary. 呼吸計測の酸化的リン酸化の解析と組み合わせてサポニン透過処理繊維の調製物は、ミトコンドリアの機能の統合的な評価を提供します。生理的および病的状態におけるミトコンドリア呼吸はミトコンドリアの相互作用、形態および生化学を含む様​​々な規制の影響を反映することができます。