a substrate located in an energetic - beam instrument has a region of interest to be extracted as a sample for further analysis . cuts are made in the substrate to define a sample , and a stress - buffer layer is formed over the region of interest or adjacent to it . an isolating cut is made to separate the portion of the substrate containing the region of interest from the bulk substrate ; however , the isolated area remains attached to the stress - buffer layer . an end - effector , such as the probe of a nano - manipulator , is attached to the stress - buffer layer , and the stress - buffer layer is cut to free the sample . the sample may then be attached to a holder by attachment of the stress - buffer layer thereto . thus the sample is never at the same time connected directly and rigidly to two different objects that may move relatively to one another , creating undesirable stresses in the sample .