TAILIEUCHUNG - Atomic Force Microscopy in Cell Biology Episode 1 Part 7

Tham khảo tài liệu 'atomic force microscopy in cell biology episode 1 part 7', kỹ thuật - công nghệ, cơ khí - chế tạo máy phục vụ nhu cầu học tập, nghiên cứu và làm việc hiệu quả | 106 Martin Benoit Fig. 9 Light microscopic image of a single cell on the sensor cells on the surface are out of focus and schematics of a force experiment Benoit et al. 2000 . interactions. Particularly in the last part of this deadhesion force trace the typical pattern for tether formation appears Hochmuth et al. 1996 . Adhesion of the nondeveloped cells used in this experiment is known to be Ca2 dependent Beug Katz Stein et al. 1973 . To test this Ca2 sensitivity 5 mM EDTA a chelating agent was added to the buffer. As illustrated at the bottom of Fig. 10B the adhesion is drastically reduced. Within the duration of the experiments this low amount of EDTA did not affect the cells integrity. Since the cells tend to move on the surface of the dish it is necessary to check the cell contact by the built-in light microscope and readjust the positioning of the cells. After growth-phase cells were brought together by contact forces of 30-40 pN applied for only s less than 20 of the de-adhesion traces showed binding between the cells Fig. 10A . The histogram of the deadhesion forces showed a broad distribution with a maximum at about 50 pN. The low frequency of these de-adhesion events implies that based on Poisson statistics more than 90 of the contacts should reflect single binding events. Thus the width of the force distribution most likely reflects a multitude of molecular species involved in the Ca2 -dependent adhesion. In the presence of 5 mM EDTA 96 of the cells did not establish detectable adhesion within s even when they were brought into contact with an increased force of 90 pN Fig. 10B . On the basis Fig. 10 Undeveloped cells lacking the CSA molecule express several Ca2 -dependent adhesion molecules A B . Experiments in PBS A result in a typical rupture force spectrum derived from 5760 traces inset after contact for s at 35 pN. Below a representative trace from a prolonged contact for 20 s at 150 pN. Experiments in 5 mM EDTA B result in a force .

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