Three-dimensional microscopic freezing and thawing behavior of biological tissues revealed by real-time imaging using confocal laser scanning microscopy

Three-dimensional behavior of ice crystals and cells during freezing and thawing of biological tissues was investigated microscopically in real time by using a confocal laser scanning microscope (CLSM) and a fluorescent dye. Fresh white meat of chicken was stained in physiological saline, and then frozen and thawed under two different thermal protocols. The CLSM noninvasively produced tomograms of the tissues to clarify the pattern of freezing, the morphology of ice crystals in the tissues, and the interaction between ice crystals and cells. The results were compared and correlated with observed histological changes in the post-thaw tissues.