Optical density at 600 nm (OD₆₀₀) measurements are routinely and quickly taken to estimate cell density in cultivation and to track cell growth. The yeast Saccharomyces cerevisiae is one of the microorganisms most used in industry, and the OD₆₀₀ values are frequently adopted as the indicator of yeast cell density, according to the Beer–Lambert law. Because the OD₆₀₀ value is based on turbidity measurement, the Beer–Lambert law can be applied only for microbial cultivation with low cell densities. The proportionality constants strongly depend on several parameters such as cell size. Typically, yeast strains are categorized into haploids and diploids. It is well known that cell size of diploid yeasts is larger than haploid cells. Additionally, polyploid (especially triploid and tetraploid) yeast cells are also employed in several human-activities such as bread-making and lager-brewing. As a matter of fact, there is almost no attention paid to the difference in the proportionality constants depending on the yeast ploidy. This study presents information for cell size of haploid, diploid, triploid, and tetraploid yeasts with isogenic background, and describes their proportionality constants (k) corresponding to the molar extinction coefficient (ε) in the Beer–Lambert law. Importantly, it was found that the constants are inversely proportional to apparent cell diameters estimated by flow cytometric analysis. Although each cell property highly depends on genetic and environmental factors, a set of results obtained from yeast strains with different ploidy in the current study would serve as a major reference source for researchers and technical experts.