Fig. 8

Illustration of preparation and observation of protoplast using LSCM. Directly mounting the samples between the cover slide and the glass slide is not preferred, as it may cause the cells to be compressed due to the weight of the cover glass or excessive downward movement of the objective lens. To avoid this, a “bridge” made by cover glasses or a confocal petri dish are employed to protect samples (1). The “bridging” method is suitable for inspection under an upright microscope or an inverted microscope, or a confocal dish can be used under an inverted microscope (2). Protoplast samples were mounted, examined and screened based on their appearance under multiple fluorescence channels and bright field conditions (3). Next, selected protoplasts are imaged using lasers as the excitation source (4)