Application of Atomic Force Microscope in Imaging Ciliary Structures of Tetrahymena Thermophilia
American College of Clinical Pharmacology Annual Meeting
Statement of Purpose, Innovation or Hypothesis: Cilia are antenna-like structures that are considered as useless evolutionary vestige, similar to tailbone. But recent research indicates that cilia are cellular organelles that have functional significance in maintaining health. Ciliary dysfunctions have been implicated in a number of clinical disorders like polycystic kidney disease and are now grouped together as ciliopathies. As tetrahymena thermophilia is a highly ciliated single celled protozoan, it was taken as an experimental model to characterize cilia using Atomic Force Microscope.
Description of Methods and Materials: Tetrahymena cells were grown at 25 degree centigrade in the axenic medium of Dentler without shaking for one week. Cells were then collected and washed for three times with PBS buffer. Then 1uL of cells were fixed on to the glass slide and placed in low temperature incubator at 15 degree centigrade until imaging is completed.
Data and Results: Atomic Force microscope with a scan head for liquid measurements is used for imaging these fixed cells. On the glass slide, a total of 100 μm area along with a 0.01nm z drive resolution were scanned and the images of cell membrane were captured at 390nm, 195nm, 98nm and 39nm resolutions.
Interpretation, Conclusion or Significance: The results show the structure of cell membrane interspersed with transmembrane proteins and protruding ciliary structures. Further studies are being conducted to quantify the structural features of cilia in order to evaluate the use of tetrahymena as model organism for ciliopathies.
Tetrahymena, ciliary structures
Woo, Kyong Ha; Chen, Xidong; and Injeti, Elisha R., "Application of Atomic Force Microscope in Imaging Ciliary Structures of Tetrahymena Thermophilia" (2011). Pharmaceutical Sciences Faculty Presentations. 20.