Pharmaceutical Sciences Faculty Presentations

Development of a Method for Imaging Erythrocytes Using Atomic Force Microscope

Document Type

Poster Session

Event Date

9-2012

Conference/Event

American College of Clinical Pharmacology Annual Meeting

Location

San Diego, CA

Abstract

Statement of Purpose, Innovation or Hypothesis: Imaging living or fixed cells in their natural environment has been the focus of nano-biotechnology research involving Atomic Force Microscope (AFM). Because of its superior sensitivity in measuring molecular forces with accuracy, the AFM has emerged as a powerful tool for investigating structure to function related properties of cells. These studies require specific sample preparation techniques that allow the AFM tip to scan the surface topography of a cell. Given the complexities of preparing the sample under optimal conditions, the current study investigated a method for imaging erythrocytes in their native state using AFM.

Description of Methods and Materials: For this purpose, blood from healthy human volunteer is collected with sodium citrate and stored at -20 degree Centigrade until used. Erythrocytes are separated by centrifuging at 13200 rpm for 14 minutes at 4 degree Centigrade. Then they are washed with normal saline and plated on a clean glass slide and allowed to stabilize at room temperature. A NanoScience EasyScan Flex AFM system with a scan head for liquid measurements is used for imaging these erythrocytes.

Data and Results: On the glass slide, a total of 50 µm, 20 µm and 10 µm area were scanned and the images of erythrocytes were captured at different resolutions. These images will be presented that were nei-ther deformed nor ruptured due to the interactions between AFM tip and erythrocytes.

Interpretation, Conclusion or Significance: As the erythrocyte membrane is unique in its biconcave shape that is retained by its cyto-skeleton, high resolution images by AFM may play an important role in clinical differential diagnosis as well as understanding the interactions between cytoskeleton and cell membranes.

Keywords

Erythrocytes, atomic force microscope

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