Ultrastructural analyses of intramembranous particles of flowing erythrocytes and their membrane skeletal structures by in vivo cryotechnique

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We frozen living animal organs in vivo to analyze tissues and cells without ischemia and anoxia for deep-etching method and scanning electron microscopy (SEM). Mouse kidney organs were frozen, and replica membranes of flowing erythrocytes were prepared, showing their extension along long axis, and intramembranous particles were linearly arranged. By SEM, they took ellipsoidal shapes in aorta, but were discoid-shaped in inferior vena cava. In liver sinusoids, they presented variously shaped images, being changed into discoid-shapes due to cardiac arrest. They were immunostained in alveolar capillaries for membrane skeletal proteins, showing irregular shapes. They were often adhered to blood vessel endothelium, showing dynamic images which efficiently performed gas exchange. We also performed the cardiac arrest to check hemodynamics of lungs and small intestines with quantum dot injection, and microenvironments of living animal organs were involved in morphofunctions of erythrocytes.