The lens is a transparent ellipsoid organ in the anterior chamber of the eye, and is responsible for fine focusing of light onto the retina to create a clear image. The function of the lens relies on its biomechanical properties, complex interdigitations between the lens fiber cells has recently been shown to be important for lens stiffness. While the specialized morphology of lens fiber cells has been previously described by electron microscopy, little is known about the proteins that are required for these interlocking membranes.
Previous studies have relied on lens tissue sections, which do not allow clear visualization of complex cell architecture. We have created and perfected a novel technique to faithfully preserve the complex membrane interdigitations between lens fiber cells for staining and confocal microscopy imaging of specific proteins. In this protocol, there is also a new method for staining the fiber cells from the center of the lens, which is also known as the lens nucleus.
This method opens the door to understanding fiber cell differentiation and maturation by studying the mechanisms of formation and changes in membrane interdigitations.
