The Nuclear Envelope by Sue Shackleton Philippe Collas & Eric C. Schirmer

Author:Sue Shackleton, Philippe Collas & Eric C. Schirmer
Language: eng
Format: epub
Publisher: Springer New York, New York, NY

10.The same needle may be used for multiple cells and multiple dishes; however, it should be changed in the case of damage or debris stuck to the needle (see Note 7 ).

11.Once the cells are in view at 40×, change the controls to “Fine” and “Axial” (for the Eppendorf InjectMan NI 2 micromanipulator ) to enable precise control over the microneedle movement in the axial direction.

12.Try to select interphase cells that resemble a sunny-side-up egg , with the nucleus as close to the center as possible. Cells with nuclei close to the cell periphery often behave inconsistently in response to microneedle manipulation. For the analysis, it is also necessary to have a minimum of 5 μm of visible cytoplasm beyond the far side of the nucleus. Once again, substrate patterning approaches may help to increase throughput and reduce experimental variability.

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