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A microfluidic device for the hydrodynamic immobilisation of living fission yeast cells for super-resolution imaging.

Bell L, Seshia A, Lando D, Laue E, Palayret M, Lee SF, Klenerman D - Sens Actuators B Chem (2014)

Bottom Line: We describe a microfluidic device designed specifically for the reversible immobilisation of Schizosaccharomyces pombe (Fission Yeast) cells to facilitate live cell super-resolution microscopy.The novel flow design captures and holds cells in a well-defined array with minimal effect on the normal growth kinetics.Cells are held over several hours and can continue to grow and divide within the device during fluorescence imaging.

View Article: PubMed Central - PubMed

Affiliation: Cambridge Nanoscience Centre, 11 J J Thomson Avenue, Cambridge CB3 0FF, United Kingdom.

ABSTRACT

We describe a microfluidic device designed specifically for the reversible immobilisation of Schizosaccharomyces pombe (Fission Yeast) cells to facilitate live cell super-resolution microscopy. Photo-Activation Localisation Microscopy (PALM) is used to create detailed super-resolution images within living cells with a modal accuracy of >25 nm in the lateral dimensions. The novel flow design captures and holds cells in a well-defined array with minimal effect on the normal growth kinetics. Cells are held over several hours and can continue to grow and divide within the device during fluorescence imaging.

No MeSH data available.


Related in: MedlinePlus

Simulation of the trapping of a cell. (A) Pressure. Flow through a plug channel is largely blocked when a cell enters, immediately setting up a large pressure difference which holds it firmly in place. (B) Flow velocity. The reduction in flow through the plug makes it less likely that another cell will enter the same well and become trapped.
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fig0015: Simulation of the trapping of a cell. (A) Pressure. Flow through a plug channel is largely blocked when a cell enters, immediately setting up a large pressure difference which holds it firmly in place. (B) Flow velocity. The reduction in flow through the plug makes it less likely that another cell will enter the same well and become trapped.

Mentions: Fig. 3 shows the change in pressure and flow velocity caused by a sphere 5 μm in diameter partially blocking the plug channel of the first available site. Fig. 3(A) shows the pressure, which holds the cell in place without stopping the passing flow in the main channel. Fig. 3(B) shows a much reduced (although non-zero) flow through the plug channel beneath the trapped cell, with a minor increase in the flow through the neighbouring basin. This suggests that the design tends towards exclusivity in trapping one cell per basin. The depth of the basin has little effect on its efficiency, as long as it is deep enough to shield the trapped cell from the main flow. This reduces any forces that might act to either move the cell or cause unnecessary shear stress on it – a feature which is lacking in many pre-existing similar cell trapping designs [17,18].


A microfluidic device for the hydrodynamic immobilisation of living fission yeast cells for super-resolution imaging.

Bell L, Seshia A, Lando D, Laue E, Palayret M, Lee SF, Klenerman D - Sens Actuators B Chem (2014)

Simulation of the trapping of a cell. (A) Pressure. Flow through a plug channel is largely blocked when a cell enters, immediately setting up a large pressure difference which holds it firmly in place. (B) Flow velocity. The reduction in flow through the plug makes it less likely that another cell will enter the same well and become trapped.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4375559&req=5

fig0015: Simulation of the trapping of a cell. (A) Pressure. Flow through a plug channel is largely blocked when a cell enters, immediately setting up a large pressure difference which holds it firmly in place. (B) Flow velocity. The reduction in flow through the plug makes it less likely that another cell will enter the same well and become trapped.
Mentions: Fig. 3 shows the change in pressure and flow velocity caused by a sphere 5 μm in diameter partially blocking the plug channel of the first available site. Fig. 3(A) shows the pressure, which holds the cell in place without stopping the passing flow in the main channel. Fig. 3(B) shows a much reduced (although non-zero) flow through the plug channel beneath the trapped cell, with a minor increase in the flow through the neighbouring basin. This suggests that the design tends towards exclusivity in trapping one cell per basin. The depth of the basin has little effect on its efficiency, as long as it is deep enough to shield the trapped cell from the main flow. This reduces any forces that might act to either move the cell or cause unnecessary shear stress on it – a feature which is lacking in many pre-existing similar cell trapping designs [17,18].

Bottom Line: We describe a microfluidic device designed specifically for the reversible immobilisation of Schizosaccharomyces pombe (Fission Yeast) cells to facilitate live cell super-resolution microscopy.The novel flow design captures and holds cells in a well-defined array with minimal effect on the normal growth kinetics.Cells are held over several hours and can continue to grow and divide within the device during fluorescence imaging.

View Article: PubMed Central - PubMed

Affiliation: Cambridge Nanoscience Centre, 11 J J Thomson Avenue, Cambridge CB3 0FF, United Kingdom.

ABSTRACT

We describe a microfluidic device designed specifically for the reversible immobilisation of Schizosaccharomyces pombe (Fission Yeast) cells to facilitate live cell super-resolution microscopy. Photo-Activation Localisation Microscopy (PALM) is used to create detailed super-resolution images within living cells with a modal accuracy of >25 nm in the lateral dimensions. The novel flow design captures and holds cells in a well-defined array with minimal effect on the normal growth kinetics. Cells are held over several hours and can continue to grow and divide within the device during fluorescence imaging.

No MeSH data available.


Related in: MedlinePlus