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We investigate the effects of 1(st) order spherical aberration and defocus upon the stiffness of an optical trap tens of μm into the sample. We control both these aberrations with a spatial light modulator. The key to maintain optimum trap stiffness over a range of depths is a specific non-trivial combination of defocus and axial objective position. This optimisation increases the trap stiffness by up to a factor of 3 and allows trapping of 1 μm polystyrene beads up to 50 μm deep in the sample.

Holographic aberration correction: optimising the stiffness of an optical trap deep in the sample