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Single cell deposition and patterning with a robotic system.

Lu Z, Moraes C, Ye G, Simmons CA, Sun Y - PLoS ONE (2010)

Bottom Line: By integrating computer vision and motion control algorithms, the system visually tracks a cell in real time and controls multiple positioning devices simultaneously to accurately pick up a single cell, transfer it to a desired substrate, and deposit it at a specified location.A traditional glass micropipette is used, and whole- and partial-cell aspiration techniques are investigated to manipulate single cells.Partially aspirating cells resulted in an operation speed of 15 seconds per cell and a 95% success rate.

View Article: PubMed Central - PubMed

Affiliation: Department of Mechanical and Industrial Engineering, University of Toronto, Toronto, Ontario, Canada.

ABSTRACT
Integrating single-cell manipulation techniques in traditional and emerging biological culture systems is challenging. Microfabricated devices for single cell studies in particular often require cells to be spatially positioned at specific culture sites on the device surface. This paper presents a robotic micromanipulation system for pick-and-place positioning of single cells. By integrating computer vision and motion control algorithms, the system visually tracks a cell in real time and controls multiple positioning devices simultaneously to accurately pick up a single cell, transfer it to a desired substrate, and deposit it at a specified location. A traditional glass micropipette is used, and whole- and partial-cell aspiration techniques are investigated to manipulate single cells. Partially aspirating cells resulted in an operation speed of 15 seconds per cell and a 95% success rate. In contrast, the whole-cell aspiration method required 30 seconds per cell and achieved a success rate of 80%. The broad applicability of this robotic manipulation technique is demonstrated using multiple cell types on traditional substrates and on open-top microfabricated devices, without requiring modifications to device designs. Furthermore, we used this serial deposition process in conjunction with an established parallel cell manipulation technique to improve the efficiency of single cell capture from ∼80% to 100%. Using a robotic micromanipulation system to position single cells on a substrate is demonstrated as an effective stand-alone or bolstering technology for single-cell studies, eliminating some of the drawbacks associated with standard single-cell handling and manipulation techniques.

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Single cells deposited on microfabricated substrates.(A) Single cells deposited on the flat surface of a microfabricated array of mechanically active cell culture sites. Inset shows a magnified view of an individual unit on the culture array (deposited cell marked with an arrow). (B) Cells deposited in an array of microwells via robotically controlled micropipette manipulation.
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pone-0013542-g003: Single cells deposited on microfabricated substrates.(A) Single cells deposited on the flat surface of a microfabricated array of mechanically active cell culture sites. Inset shows a magnified view of an individual unit on the culture array (deposited cell marked with an arrow). (B) Cells deposited in an array of microwells via robotically controlled micropipette manipulation.

Mentions: On a flat culture substrate, such as the array of mechanically active cell culture regions on the microdevice shown in Figure 3A, cells could be positioned within 10 µm of each other. Single cells were successfully patterned on all units of the array (Figure 3A). Cells were also successfully patterned into a more topographically complex substrate, an array of microfabricated wells (Figure 3B). Viability was confirmed using a calcein AM stain.


Single cell deposition and patterning with a robotic system.

Lu Z, Moraes C, Ye G, Simmons CA, Sun Y - PLoS ONE (2010)

Single cells deposited on microfabricated substrates.(A) Single cells deposited on the flat surface of a microfabricated array of mechanically active cell culture sites. Inset shows a magnified view of an individual unit on the culture array (deposited cell marked with an arrow). (B) Cells deposited in an array of microwells via robotically controlled micropipette manipulation.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC2958835&req=5

pone-0013542-g003: Single cells deposited on microfabricated substrates.(A) Single cells deposited on the flat surface of a microfabricated array of mechanically active cell culture sites. Inset shows a magnified view of an individual unit on the culture array (deposited cell marked with an arrow). (B) Cells deposited in an array of microwells via robotically controlled micropipette manipulation.
Mentions: On a flat culture substrate, such as the array of mechanically active cell culture regions on the microdevice shown in Figure 3A, cells could be positioned within 10 µm of each other. Single cells were successfully patterned on all units of the array (Figure 3A). Cells were also successfully patterned into a more topographically complex substrate, an array of microfabricated wells (Figure 3B). Viability was confirmed using a calcein AM stain.

Bottom Line: By integrating computer vision and motion control algorithms, the system visually tracks a cell in real time and controls multiple positioning devices simultaneously to accurately pick up a single cell, transfer it to a desired substrate, and deposit it at a specified location.A traditional glass micropipette is used, and whole- and partial-cell aspiration techniques are investigated to manipulate single cells.Partially aspirating cells resulted in an operation speed of 15 seconds per cell and a 95% success rate.

View Article: PubMed Central - PubMed

Affiliation: Department of Mechanical and Industrial Engineering, University of Toronto, Toronto, Ontario, Canada.

ABSTRACT
Integrating single-cell manipulation techniques in traditional and emerging biological culture systems is challenging. Microfabricated devices for single cell studies in particular often require cells to be spatially positioned at specific culture sites on the device surface. This paper presents a robotic micromanipulation system for pick-and-place positioning of single cells. By integrating computer vision and motion control algorithms, the system visually tracks a cell in real time and controls multiple positioning devices simultaneously to accurately pick up a single cell, transfer it to a desired substrate, and deposit it at a specified location. A traditional glass micropipette is used, and whole- and partial-cell aspiration techniques are investigated to manipulate single cells. Partially aspirating cells resulted in an operation speed of 15 seconds per cell and a 95% success rate. In contrast, the whole-cell aspiration method required 30 seconds per cell and achieved a success rate of 80%. The broad applicability of this robotic manipulation technique is demonstrated using multiple cell types on traditional substrates and on open-top microfabricated devices, without requiring modifications to device designs. Furthermore, we used this serial deposition process in conjunction with an established parallel cell manipulation technique to improve the efficiency of single cell capture from ∼80% to 100%. Using a robotic micromanipulation system to position single cells on a substrate is demonstrated as an effective stand-alone or bolstering technology for single-cell studies, eliminating some of the drawbacks associated with standard single-cell handling and manipulation techniques.

Show MeSH
Related in: MedlinePlus