Abstract: A single-cell-squashing technique was established for the observation of organelle DNA distribution in tobacco(Nicotiana tabacum L.) zygotes. Two protocols, simultaneous or successive stain and fixation, were compared and both methods could be used as alternative for single cell observation of organelle DNA. We also tested the effect of single-cell-squashing technique combining with CCD observation and whole mount material observed by CLSM. Experiments showed both of them have their own advantages in application.