Microtome-Based Sectioning Method for Generating Thin Cell Layer Cultures in Phalaenopsis

The production of virus-free plants is crucial in the supply chain management in the international trade of orchids. Excising the shoot tip from orchid plants infected with viruses is a decisive technique for obtaining virus-free plantlets. Previous reports demonstrated that manual excision could obtain a minimum size of only approximately 300–500 μm for the shoot apex, consisting of the apical meristem and one to three leaf primordia. The advanced thin cell layer (TCL) excision technique makes obtaining the shoot apex as small as 100 μm possible, with proficient practitioners achieving excision down to 30 μm. By employing the advanced TCL excision technique, it is possible to excise the smallest shoot apex, which can be applied to generating the virus-free shoot apex with high efficiency. This chapter focuses on the techniques necessary to obtain tiny amounts of tissue to produce virus-free plantlets and regeneration studies.