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Over-mixing creates an amalgam which is hard to form and porous and prone to fracture while under-mixing may create an amalgam which is too soft with too much free mercury

Islets were then washed with the Cholesterol Detection wash buffer, incubated for a minimum of 1 hour in Filipin III diluted in the Cholesterol Detection Assay buffer, and washed again in wash buffer before immediate imaging on a Zeiss LSM-780 inverted confocal laser-scanning microscope from the Facility for Imaging by Light Microscopy (FILM) at Imperial College London, with a 63X objective and a laser excitation wavelength of 340-380 nm and emission of 385-470 nm

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This psychological benefit often matters as much as the numerical weight loss for long-term success