Place cells, spatially responsive hippocampal cells, provide the neural substrate supporting navigation and spatial memory. Historically most studies of these neurons have used electrophysiological recordings from implanted electrodes but optical methods, measuring intracellular calcium, are becoming increasingly common. Several methods have been proposed as a means to identify place cells based on their calcium activity but there is no common standard and it is unclear how reliable different approaches are. Here we tested three methods that have previously been applied to two-photon hippocampal imaging or electrophysiological data, using both model datasets and real imaging data. These methods use different parameters to identify place cells, including the peak activity in the place field, compared to other locations (the Peak method); the stability of cells’ activity over repeated traversals of an environment (Stability method); and a combination of these parameters with the size of the place field (Combination method). The three methods performed differently from each other on both model and real data. The Peak method showed high sensitivity and specificity for detecting model place cells and was the most robust to variations in place field width, reliability and field location. In real datasets, vastly different numbers of place cells were identified using the three methods, with little overlap between the populations identified as place cells. Therefore, choice of place cell detection method dramatically affects the number and properties of identified cells. We recommend the Peak method be used in future studies to identify place cell populations, unless there is an explicit theoretical reason for detecting cells with more narrowly defined properties.