Analysis of Penetration of Colloidal Carbon (India Black Ink) in Blood Clots by Light Microscopy
Mentor 1
Julie Oliver
Location
Union 240
Start Date
5-4-2019 12:00 PM
Description
Ischemic stroke occurs when the blood flow to the brain is reduced or blocked, usually by a thrombus, or blood clot. The only FDA-approved treatment for ischemic/thrombotic stroke patients is the administration of the drug tissue plasminogen activator (tPA). tPA targets fibrin, the protein component of blood clots. While it can reopen the blocked vessel, it can also prevent effective clotting elsewhere, potentially leading to excessive bleeding and brain hemorrhaging. Because of these side effects, there is great need for a new treatment. We are investigating a technique that targets platelets, the cellular component of blood clots, using fibrinogen-conjugated nanoparticles. Our goal is to develop a rapid screening procedure that will allow us to compare how these nanoparticles interact with samples from different blood donors. Clots were formed using various amount of thrombin in both platelet-rich plasma (PRP) and whole blood (WB) from three donors in our normal pool. We hypothesized that over a range of physiologically relevant doses of thrombin, we will see differences in the clot structure as a function of the dose of thrombin, as revealed by the penetration of colloidal carbon (India Black ink) into the clot as a substitute for nanoparticles. These clots were soaked in ink, frozen, cryosectioned and analyzed by light microscopy. Images of each clot were collected and analyzed in ImageJ using the threshold option. Preliminary results indicate that our procedure detects differences in clot structure, but it remains unclear whether the differences depend on the thrombin dose or on donor-to-donor variability. Testing additional blood donors will help us determine how effective this technique will be as a means of examining clot structure.
Analysis of Penetration of Colloidal Carbon (India Black Ink) in Blood Clots by Light Microscopy
Union 240
Ischemic stroke occurs when the blood flow to the brain is reduced or blocked, usually by a thrombus, or blood clot. The only FDA-approved treatment for ischemic/thrombotic stroke patients is the administration of the drug tissue plasminogen activator (tPA). tPA targets fibrin, the protein component of blood clots. While it can reopen the blocked vessel, it can also prevent effective clotting elsewhere, potentially leading to excessive bleeding and brain hemorrhaging. Because of these side effects, there is great need for a new treatment. We are investigating a technique that targets platelets, the cellular component of blood clots, using fibrinogen-conjugated nanoparticles. Our goal is to develop a rapid screening procedure that will allow us to compare how these nanoparticles interact with samples from different blood donors. Clots were formed using various amount of thrombin in both platelet-rich plasma (PRP) and whole blood (WB) from three donors in our normal pool. We hypothesized that over a range of physiologically relevant doses of thrombin, we will see differences in the clot structure as a function of the dose of thrombin, as revealed by the penetration of colloidal carbon (India Black ink) into the clot as a substitute for nanoparticles. These clots were soaked in ink, frozen, cryosectioned and analyzed by light microscopy. Images of each clot were collected and analyzed in ImageJ using the threshold option. Preliminary results indicate that our procedure detects differences in clot structure, but it remains unclear whether the differences depend on the thrombin dose or on donor-to-donor variability. Testing additional blood donors will help us determine how effective this technique will be as a means of examining clot structure.
