Abstract Details

Title A Novel Non-Injectional Method of Three-Dimensional Staining of Human Central Nervous System Microvasculature Using Cadmium

Topic Cerebrovascular Disease and Interventional Neurology

Presentation(s) P05 - (-)

Poster/Presentation
Number 239

Objective

Background BACKGROUND: Despite their advantages, there are few methods for non-injectional microvascular staining. In addition, a number of these methods are limited in their abilities for staining all segments of MVB, including arterioles, capillaries, and venules. Non-injectional methods may avoid some limitations of injectional methods such as staining closed vessels. Development of a non-injectional technique showing the entire MVB can aid in the investigation of various CNS pathologies.

Design/Methods DESIGN/METHODS: Fresh or frozen, non-fixed tissue blocks were obtained from rapidly postmortem human Alzheimer disease and control CNS. Tissues were fixed in 4% formalin for 1-2 days, followed by sectioning into 60-120 um sections under -13[deg]C. Subsequently, sections were stored in 0.9% normal saline. Sections were placed in incubation solution consisting of ammonium chloride buffer at pH 9-9.6, cadmium acetate, and adenoside triphosphate. Next, sections were transferred into a lead acetate buffered solution where cadmium phosphate precipitation formed on the endothelium was replaced by lead phosphate. Subsequently, sections were washed and transferred into sodium sulfide solution with resulting brown lead sulfide precipitate.

Results RESULTS: This method utilizing buffered cadmium solution at pH 9-9.6 allows for selective MVB staining with all of components - arterioles, capillaries, and venules. Arterioles are distinguished from venules by darker and thicker staining of their walls. Morphologically, abnormal vessels are able to be described due to adequate thickness of the sections. Glia, neurons, and other structures of brain parenchyma remain unstained allowing for utilization of thicker sections.

Conclusions CONCLUSIONS: This novel cadmium staining technique selectively stains MVB and may have a role in studying morphological changes in microvasculature in a wide range of brain disease, including but not limited to stroke and neoplasm.

Authors/Disclosures
Amaiak Chilingaryan, MD (CA) PRESENTER	No disclosure on file
Jacqueline Montes, PT, EdD, NCS (Columbia University Medical Center)	Ms. Montes has received personal compensation in the range of $10,000-$49,999 for serving on a Scientific Advisory or Data Safety Monitoring board for Biogen. Ms. Montes has received personal compensation in the range of $500-$4,999 for serving on a Scientific Advisory or Data Safety Monitoring board for F. Hoffman LaRoche. Ms. Montes has received personal compensation in the range of $500-$4,999 for serving on a Scientific Advisory or Data Safety Monitoring board for Scholar Rock. Ms. Montes has received personal compensation in the range of $500-$4,999 for serving on a Scientific Advisory or Data Safety Monitoring board for Sarepta. The institution of Ms. Montes has received research support from NIH/NICHD. The institution of Ms. Montes has received research support from Muscular Dystrophy Association. The institution of Ms. Montes has received research support from Cure SMA.
	No disclosure on file

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