The invention discloses a phenyl-substituted bodipy and diphenylamine fluorene based two-photon fluorescent dye. A structural general formula of the fluorescent dye is represented by a formula shown in the description, wherein R is C1-C18 alkyl. Steps for synthesizing the fluorescent dye are as follows: producing 5-phenyl-substituted and 2,6-iodinated bodipy from 2,4-dimethyl pyrrole and benzaldehyde, which serve as raw materials; subjecting fluorene, which serves as a raw material, to bromination, alkylation, Pd(0) catalyzed amination, a Pd(0) and CuI catalyzed Sonogashira cross-coupling reaction and trimethylsilyl removal, so as to produce a diphenylamine-fluorene-acetylene compound; and subjecting the 2,6-iodinated bodipy and the diphenylamine-fluorene-acetylene compound to a reaction, thereby obtaining the target product. The target compound has relatively high two-photon fluorescence performance, the maximum two-photon absorption cross section in toluene reaches 265GM, and the fluorescent quantum yield is 0.35, so that a new design way of think is provided for the synthesis and application of bodipy-based two-photon fluorescent dyes.