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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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Related in: MedlinePlus

Multiple cell types deposited on the same substrate.Fluorescently labeled endothelial cells (Green) and fibroblasts (Red) are deposited in a pattern, demonstrating the ability to precisely manipulate multiple cell types on a single substrate.
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pone-0013542-g004: Multiple cell types deposited on the same substrate.Fluorescently labeled endothelial cells (Green) and fibroblasts (Red) are deposited in a pattern, demonstrating the ability to precisely manipulate multiple cell types on a single substrate.

Mentions: A critical advantage of this microrobotic system is the ability to manipulate multiple cell types on a single substrate. This is extremely challenging to do using conventional technologies. For this demonstration, we filled an array of microwells using the robotic system, with two cell types. Figure 4 shows fibroblasts and endothelial cells robotically deposited to form a ‘UT’ (University of Toronto) pattern in individual microwells, from which they can be transferred onto any surface using the BioFlipChip method [13]. The ability to manipulate multiple cell types can be used to answer specific biological questions, such as those related to co-culture and interacting effects between multiple cell types.


Single cell deposition and patterning with a robotic system.

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

Multiple cell types deposited on the same substrate.Fluorescently labeled endothelial cells (Green) and fibroblasts (Red) are deposited in a pattern, demonstrating the ability to precisely manipulate multiple cell types on a single substrate.
© Copyright Policy
Related In: Results  -  Collection

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

pone-0013542-g004: Multiple cell types deposited on the same substrate.Fluorescently labeled endothelial cells (Green) and fibroblasts (Red) are deposited in a pattern, demonstrating the ability to precisely manipulate multiple cell types on a single substrate.
Mentions: A critical advantage of this microrobotic system is the ability to manipulate multiple cell types on a single substrate. This is extremely challenging to do using conventional technologies. For this demonstration, we filled an array of microwells using the robotic system, with two cell types. Figure 4 shows fibroblasts and endothelial cells robotically deposited to form a ‘UT’ (University of Toronto) pattern in individual microwells, from which they can be transferred onto any surface using the BioFlipChip method [13]. The ability to manipulate multiple cell types can be used to answer specific biological questions, such as those related to co-culture and interacting effects between multiple cell types.

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