Cell culture arrays using magnetic force-based cell patterning for dynamic single cell analysis

Abstract

In order to understand the behavior of individual cells, single cell analyses have attracted attention since most cell-based assays provide data with values averaged across a large number of cells. Techniques for the manipulation and analysis of single cells are crucial for understanding the behavior of individual cells. In the present study, we have developed single cell culture arrays using magnetic force and a pin holder, which enables the allocation of the magnetically labeled cells on arrays, and have analyzed their dynamics. The pin holder was made from magnetic soft iron and contained more than 6000 pillars on its surface. The pin holder was placed on a magnet to concentrate the magnetic flux density above the pillars. NIH/3T3 fibroblasts that were labeled with magnetite cationic liposomes (MCLs) were seeded into a culture dish, and the dish was placed over the pin holder with the magnet. The magnetically labeled cells were guided on the surface where the pillars were positioned and allocated on the arrays with a high resolution. Single-cell patterning was achieved by adjusting the number of cells seeded, and the target cell was collected by a micromanipulator after removing the pin holder with the magnet. Furthermore, change in the morphology of magnetically patterned cells was analyzed by microscopic observation, and cell spreading on the array was observed with time duration. Magnetic force-based cell patterning on cell culture arrays would be a suitable technique for the analysis of cell behavior in studies of cell-cell variation and cell-cell interactions.

Cellular self-organization on micro-structured surfaces

“…The number of trapped cells per adhesive spot follows a Poisson distribution. [25][26][27][28] As a result, such arrays exhibit a broad distribution of cell numbers on the adhesive islands. For an expected value of l ¼ 1 (one cell per site), the maximal single-cell occupancy achievable is 37%.…”

Exploitation of physical and chemical constraints for three-dimensional microtissue construction in microfluidics

“…In the third type of trapping, magnetic particles are selectively attached to cells and used in microfluidic devices for rare cell types separation, 88,89 and recently, in tissue engineering 49,90 ͓Fig. 2͑f͔͒.…”

“…Ino et al 90 devised cell culture arrays using magnetic patterning and utilizing magnetic cationic liposomes for dynamic single cell analysis. For this application, gradients of the magnetic field were achieved using a pin holder and used for a 3D cell culture array.…”

Identification of antigen‐specific B cells by concurrent monitoring of intracellular Ca2+ mobilization and antigen binding with microwell array chip system equipped with a CCD imager

“…After that, the MAC-CCD system detected the position of each microwell on the chip again. Fifteen seconds after the stimulation, the fluorescence images of the fluo-4 signals were acquired on the whole microwell area every 3 s for an additional 45 s (scanning points [6][7][8][9][10][11][12][13][14][15][16][17][18][19][20]. The cells inside the wells did not move out of the wells after four gentle washes.…”