Abstract：
BACKGROUND: This study aimed to develop a modified histochemical staining technique to successfully identify arterial and venous segments of brain microvessels.
METHODS: Gelatin/red ink-alkaline phosphatase-oil red O (GIAO) staining was developed from the traditional gelatin-ink perfusion method. Oil red Chinese ink for brush writing and painting mixed with gelatin was used to label cerebral vascular lumens. Subsequently, alkaline phosphatase staining was used to label endothelial cells on the arterial segments of cerebral microvessels. Thereafter, the red ink color in vessel lumens was highlighted with oil red O staining.
RESULTS: The arterial segments of the brain microvessels exhibited red lumens surrounded by dark blue walls, while the venous segments were bright red following GIAO staining. Meanwhile, the nerve fiber bundles were stained brownish-yellow, and the nuclei appeared light green under light microscope. After cerebral infarction, we used GIAO staining to determine angiogenesis features and detected notable vein proliferation inside the infarct core. Moreover, GIAO staining in conjunction with hematoxylin staining was performed to assess the infiltration of foamy macrophages.
CONCLUSIONS: Red Chinese ink enabled subsequent multiple color staining on brain section. Oil red O was introduced to improved the resolution and contrast between arterial and venous segments of microvessels.
CONCLUSIONS: With excellent resolution, GIAO staining effectively distinguished arterial and venous segments of microvessels in both normal and ischemic brain tissue. GIAO staining, as described in the present study, will be useful for histological investigations of microvascular bed alterations in a variety of brain disorders.
METHODS: Gelatin/red ink-alkaline phosphatase-oil red O (GIAO) staining was developed from the traditional gelatin-ink perfusion method. Oil red Chinese ink for brush writing and painting mixed with gelatin was used to label cerebral vascular lumens. Subsequently, alkaline phosphatase staining was used to label endothelial cells on the arterial segments of cerebral microvessels. Thereafter, the red ink color in vessel lumens was highlighted with oil red O staining.
RESULTS: The arterial segments of the brain microvessels exhibited red lumens surrounded by dark blue walls, while the venous segments were bright red following GIAO staining. Meanwhile, the nerve fiber bundles were stained brownish-yellow, and the nuclei appeared light green under light microscope. After cerebral infarction, we used GIAO staining to determine angiogenesis features and detected notable vein proliferation inside the infarct core. Moreover, GIAO staining in conjunction with hematoxylin staining was performed to assess the infiltration of foamy macrophages.
CONCLUSIONS: Red Chinese ink enabled subsequent multiple color staining on brain section. Oil red O was introduced to improved the resolution and contrast between arterial and venous segments of microvessels.
CONCLUSIONS: With excellent resolution, GIAO staining effectively distinguished arterial and venous segments of microvessels in both normal and ischemic brain tissue. GIAO staining, as described in the present study, will be useful for histological investigations of microvascular bed alterations in a variety of brain disorders.
摘要：
背景：这项研究旨在开发一种改良的组织化学染色技术，以成功鉴定脑微血管的动脉和静脉段。
方法：明胶/红墨水-碱性磷酸酶-油红O（GIAO）染色是从传统的明胶-墨水灌注法发展而来的。油红中国笔墨和明胶混合用于标记脑血管管腔。随后，碱性磷酸酶染色用于标记脑微血管动脉段上的内皮细胞。此后,用油红O染色突出显示血管腔中的红色墨水颜色。
结果：脑微血管的动脉段显示出被深蓝色壁包围的红色管腔，而静脉节段在GIAO染色后呈鲜红色。同时,神经纤维束被染成棕黄色，在光学显微镜下原子核呈浅绿色。脑梗塞后，我们使用GIAO染色来确定血管生成特征,并检测到梗死核心内明显的静脉增生.此外，进行GIAO染色结合苏木精染色以评估泡沫巨噬细胞的浸润。
结论：红色中国墨水能够在大脑切片上进行随后的多色染色。引入油红O以提高微血管的动脉和静脉段之间的分辨率和对比度。
结论：具有出色的分辨率，GIAO染色可有效区分正常和缺血脑组织中微血管的动脉和静脉段。GIAO染色,如本研究中所述，对于各种脑部疾病的微血管床改变的组织学研究很有用。
方法：明胶/红墨水-碱性磷酸酶-油红O（GIAO）染色是从传统的明胶-墨水灌注法发展而来的。油红中国笔墨和明胶混合用于标记脑血管管腔。随后，碱性磷酸酶染色用于标记脑微血管动脉段上的内皮细胞。此后,用油红O染色突出显示血管腔中的红色墨水颜色。
结果：脑微血管的动脉段显示出被深蓝色壁包围的红色管腔，而静脉节段在GIAO染色后呈鲜红色。同时,神经纤维束被染成棕黄色，在光学显微镜下原子核呈浅绿色。脑梗塞后，我们使用GIAO染色来确定血管生成特征,并检测到梗死核心内明显的静脉增生.此外，进行GIAO染色结合苏木精染色以评估泡沫巨噬细胞的浸润。
结论：红色中国墨水能够在大脑切片上进行随后的多色染色。引入油红O以提高微血管的动脉和静脉段之间的分辨率和对比度。
结论：具有出色的分辨率，GIAO染色可有效区分正常和缺血脑组织中微血管的动脉和静脉段。GIAO染色,如本研究中所述，对于各种脑部疾病的微血管床改变的组织学研究很有用。
