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We evaluated statistical models for our multiplex immunofluorescence study of T cell subset densities in colorectal cancer. Using 1235 cases, we compared 7 models\u2014ordinal logistic regression, Poisson, quasi-Poisson, quadratic negative binomial (NB), linear NB, zero-inflated NB, and hurdle NB models\u2014assessing associations with a strong (microsatellite instability, MSI) and a weak (calcium intake) exposure. Simulation studies assessed type I error and power. Effect estimates were generally consistent for the strong exposure (MSI) but varied for the weaker exposure (calcium). Simulations revealed inflated false-positive rates for the Poisson and NB-based models, including quadratic NB, zero-inflated, and hurdle, but not for ordinal logistic regression or the linear NB model. The quasi-Poisson model showed modest inflation of low P-values, but the overall P-value distribution remained approximately uniform under the null. 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