AbstractFlow cytometry is a high‐throughput tool for determining microbial abundance in a range of medical, environmental, and food‐related samples. For wine, determining the abundance of Saccharomyces cerevisiae is well‐defined and reliable. However, for the most common wine bacterium, Oenococcus oeni, using flow cytometry to determine cell concentration poses some challenges. O. oeni most often occurs in doublets or chains of varying lengths that can be greater than seven cells. This wine bacterium is also small, at 0.2–0.6 μm and may exhibit a range of morphologies including binary fission and aggregated complexes. This work demonstrates a straightforward approach to determining the suitability of flow cytometry for the chain‐forming bacteria, O. oeni, and considerations when using flow cytometry for the enumeration of small microorganisms (<0.5 μm). © 2020 International Society for Advancement of Cytometry