在沙棘籽油提取过程中,经粉碎干燥后的籽料颗粒是一种开放体系,脂滴以游离形式存在,采用油红O(Oil Red O,ORO)对其进行染色时,常规方法一次性滴加染液的方式以及后续的洗脱步骤均会导致脂滴迁移、过染等问题出现。研究构建了逐级染色法...在沙棘籽油提取过程中,经粉碎干燥后的籽料颗粒是一种开放体系,脂滴以游离形式存在,采用油红O(Oil Red O,ORO)对其进行染色时,常规方法一次性滴加染液的方式以及后续的洗脱步骤均会导致脂滴迁移、过染等问题出现。研究构建了逐级染色法,采用少量多次滴加染液的方式,并优化了染色间隔时间为1.5 min,染色级数和染液浓度可根据样品的实际油含量进行调整,因而能够客观地显影脂滴在籽料颗粒内的分布状态。以此构建的面积占比法可用于精确测定籽料中的油脂含量,其准确度与萃取质量法相当,两者的相关系数为0.998,并具有较高重现性,相对标准偏差约为2.49%。展开更多
目的比较2种神经髓鞘染色方法在Cuprizone小鼠模型脑组织中脱髓鞘的形态学变化,为实验诊断提供病理依据。方法将Cuprizone掺入饲料中喂食小鼠4周,造成神经脱髓鞘模型。取脑后切片,分别用牢固蓝(Luxol Fast Blue)和油红O两种染色方法观...目的比较2种神经髓鞘染色方法在Cuprizone小鼠模型脑组织中脱髓鞘的形态学变化,为实验诊断提供病理依据。方法将Cuprizone掺入饲料中喂食小鼠4周,造成神经脱髓鞘模型。取脑后切片,分别用牢固蓝(Luxol Fast Blue)和油红O两种染色方法观察脑纹状体和海马两个切面胼胝体髓鞘脱失变化。结果牢固蓝和油红O两种染色方法均可显示Cuprizone模型小鼠脑内胼胝体部位有脱髓鞘发生,但在纹状体切面油红O染色法能清晰显示髓鞘脱失和炎性细胞浸润,而牢固蓝染色法显示髓鞘脱失与周围组织结构界限不明显。结论与牢固蓝染色法相比,油红O染色法能更清晰显示脱髓鞘,颜色对比明显,是一种简单易行的神经髓鞘染色方法。展开更多
AIM: To develop a method for studying myocardial area at risk(AAR) in ischemic heart disease in correlation with cardiac magnetic resonance imaging(c MRI). METHODS: Nine rabbits were anesthetized, intubated and subjec...AIM: To develop a method for studying myocardial area at risk(AAR) in ischemic heart disease in correlation with cardiac magnetic resonance imaging(c MRI). METHODS: Nine rabbits were anesthetized, intubated and subjected to occlusion and reperfusion of the left circumflex coronary artery(LCx) to induce myocardial infarction(MI). ECG-triggered c MRI with delayed en-hancement was performed at 3.0 T. After euthanasia, the heart was excised with the LCx re-ligated. Bifunctional staining was performed by perfusing the aorta with a homemade red-iodized-oil(RIO) dye. The heart was then agar-embedded for ex vivo magnetic resonance imaging and sliced into 3 mm-sections. The AAR was defined by RIO-staining and digital radiography(DR). The perfusion density rate(PDR) was derived from DR for the AAR and normal myocardium. The MI was measured by in vivo delayed enhancement(i DE) and ex vivo delayed enhancement(e DE) c MRI. The AAR and MI were compared to validate the bifunctional straining for cardiac imaging research. Linear regression with Bland-Altman agreement, one way-ANOVA with Bonferroni's multiple comparison, and paired t tests were applied for statistics.RESULTS: All rabbits tolerated well the surgical procedure and subsequent c MRI sessions. The openchest occlusion and close-chest reperfusion of the LCx, double suture method and bifunctional staining were successfully applied in all animals. The percentage MI volumes globally(n = 6) and by slice(n = 25) were 36.59% ± 13.68% and 32.88% ± 12.38% on i DE, and 35.41% ± 12.25% and 32.40% ± 12.34% on e DE. There were no significant differences for MI determination with excellent linear regression correspondence(r global = 0.89; r slice = 0.9) between i DE and e DE. The percentage AAR volumes globally(n = 6) and by slice(n = 25) were 44.82% ± 15.18% and 40.04% ± 13.64% with RIO-staining, and 44.74% ± 15.98% and 40.48% ± 13.26% by DR showing high correlation in linear regression analysis(r global = 0.99; r slice = 1.0). The mean differences of the two AAR measurements on BlandAltman were almost zero, indicating RIO-staining and DR were essentially equivalent or inter-replaceable. The AAR was significantly larger than MI both globally and slice-by-slice(P < 0.01). After correction with the background and the blank heart without bifunctional staining(n = 3), the PDR for the AAR and normal myocardium was 32% ± 15% and 35.5% ± 35%, respectively,which is significantly different(P < 0.001), suggesting that blood perfusion to the AAR probably by collateral circulation was only less than 10% of that in the normal myocardium.CONCLUSION: The myocardial area at risk in ischemic heart disease could be accurately determined postmortem by this novel bifunctional staining, which may substantially contribute to translational cardiac imaging research.展开更多
文摘在沙棘籽油提取过程中,经粉碎干燥后的籽料颗粒是一种开放体系,脂滴以游离形式存在,采用油红O(Oil Red O,ORO)对其进行染色时,常规方法一次性滴加染液的方式以及后续的洗脱步骤均会导致脂滴迁移、过染等问题出现。研究构建了逐级染色法,采用少量多次滴加染液的方式,并优化了染色间隔时间为1.5 min,染色级数和染液浓度可根据样品的实际油含量进行调整,因而能够客观地显影脂滴在籽料颗粒内的分布状态。以此构建的面积占比法可用于精确测定籽料中的油脂含量,其准确度与萃取质量法相当,两者的相关系数为0.998,并具有较高重现性,相对标准偏差约为2.49%。
文摘目的比较2种神经髓鞘染色方法在Cuprizone小鼠模型脑组织中脱髓鞘的形态学变化,为实验诊断提供病理依据。方法将Cuprizone掺入饲料中喂食小鼠4周,造成神经脱髓鞘模型。取脑后切片,分别用牢固蓝(Luxol Fast Blue)和油红O两种染色方法观察脑纹状体和海马两个切面胼胝体髓鞘脱失变化。结果牢固蓝和油红O两种染色方法均可显示Cuprizone模型小鼠脑内胼胝体部位有脱髓鞘发生,但在纹状体切面油红O染色法能清晰显示髓鞘脱失和炎性细胞浸润,而牢固蓝染色法显示髓鞘脱失与周围组织结构界限不明显。结论与牢固蓝染色法相比,油红O染色法能更清晰显示脱髓鞘,颜色对比明显,是一种简单易行的神经髓鞘染色方法。
基金Supported by The awarded grants of the KU Leuven Molecular Small Animal Imaging Center Mo SAIC(KUL EF/05/08)the Center of Excellence in vivo Molecular Imaging Research(IMIR)of KU Leuven+1 种基金a EU Project Asia-Link Cf P 2006-Europe Aid/123738/C/ACT/Multi-Proposal,No.128-498/111Jiangsu Province Natural Science Foundation,China,No.BK2010594
文摘AIM: To develop a method for studying myocardial area at risk(AAR) in ischemic heart disease in correlation with cardiac magnetic resonance imaging(c MRI). METHODS: Nine rabbits were anesthetized, intubated and subjected to occlusion and reperfusion of the left circumflex coronary artery(LCx) to induce myocardial infarction(MI). ECG-triggered c MRI with delayed en-hancement was performed at 3.0 T. After euthanasia, the heart was excised with the LCx re-ligated. Bifunctional staining was performed by perfusing the aorta with a homemade red-iodized-oil(RIO) dye. The heart was then agar-embedded for ex vivo magnetic resonance imaging and sliced into 3 mm-sections. The AAR was defined by RIO-staining and digital radiography(DR). The perfusion density rate(PDR) was derived from DR for the AAR and normal myocardium. The MI was measured by in vivo delayed enhancement(i DE) and ex vivo delayed enhancement(e DE) c MRI. The AAR and MI were compared to validate the bifunctional straining for cardiac imaging research. Linear regression with Bland-Altman agreement, one way-ANOVA with Bonferroni's multiple comparison, and paired t tests were applied for statistics.RESULTS: All rabbits tolerated well the surgical procedure and subsequent c MRI sessions. The openchest occlusion and close-chest reperfusion of the LCx, double suture method and bifunctional staining were successfully applied in all animals. The percentage MI volumes globally(n = 6) and by slice(n = 25) were 36.59% ± 13.68% and 32.88% ± 12.38% on i DE, and 35.41% ± 12.25% and 32.40% ± 12.34% on e DE. There were no significant differences for MI determination with excellent linear regression correspondence(r global = 0.89; r slice = 0.9) between i DE and e DE. The percentage AAR volumes globally(n = 6) and by slice(n = 25) were 44.82% ± 15.18% and 40.04% ± 13.64% with RIO-staining, and 44.74% ± 15.98% and 40.48% ± 13.26% by DR showing high correlation in linear regression analysis(r global = 0.99; r slice = 1.0). The mean differences of the two AAR measurements on BlandAltman were almost zero, indicating RIO-staining and DR were essentially equivalent or inter-replaceable. The AAR was significantly larger than MI both globally and slice-by-slice(P < 0.01). After correction with the background and the blank heart without bifunctional staining(n = 3), the PDR for the AAR and normal myocardium was 32% ± 15% and 35.5% ± 35%, respectively,which is significantly different(P < 0.001), suggesting that blood perfusion to the AAR probably by collateral circulation was only less than 10% of that in the normal myocardium.CONCLUSION: The myocardial area at risk in ischemic heart disease could be accurately determined postmortem by this novel bifunctional staining, which may substantially contribute to translational cardiac imaging research.
