Identification of Critical Conditions for Immunostaining in the Pea Aphid Embryos: Increasing Tissue Permeability and Decreasing Background Staining

The overall goal of this immunostaining protocol is to detect protein expression in the aphid embryos. Critical conditions are identified that increase tissue permeability and decrease background staining, both of which are long-standing problems when staining embryonic tissues of aphids. The main advantage of this technique is that it provides specific conditions for effective penetration of antibodies and reduction of background staining, which usually are not described in a standard immunostaining protocol.

We first had this idea of this measure when we were studying how germ cells are specified in a pea aphid a variety of insect model, for genomics and environmental studies. The laboratory strain of the pea aphid, a. pisum, was originally collected in central Taiwan, and has been reared on host plants for more than 300 generations.