Tissue Culture of Orchids can be done with several different types of tissue. The most common is to take a small piece from the end of an actively growing shoot. This actively growing tip will produce roots and shoots faster than other tissue types. However, there has also been research that shows the tip of a leaf will root in liquid media, this causes less damage to the stock plant than taking a shoot tip (Churchill et al., 1973).
The first step is to remove a small portion of an aerial shoot from the plant. This shoot will have nodes, internodes and is the stalk which would eventually flower. This can be done almost anytime of year, besides winter, as the orchids can be in a period of dormancy in the cooler winter months. Sterilize a scalpel by putting it in a heated glass bead sterilizer for a few seconds or by dipping it in alcohol and passing it through an alcohol lamp. Cut a healthy looking, disease free shoot off the stock plant and put it in a 10% bleach solution for 15 minutes. Then rinse the shoot in clean water three times to remove the bleach. Place the shoot on a sterile work environment, such as under a laminar flow hood to decrease the chance of contamination(Media Recommendation Guide, 2011). Sterile technique is very important in tissue culture to prevent growth of bacteria and fungi. Using the sterilized scalpel cut a 5-10 mm piece from the tip of the shoot. The rest of the shoot can also be cut into similar sized pieces depending on how many plantlets are desired. The next step will be to put this small piece of tissue (called the explant) onto the media.