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In order to prevent the spread of counterfeit and shoddy products, fluorescent fibers from inorganic fluorescent substance and organic materials are continuously being developed. In this paper, a novel method, which was different from spinning method, was presented for preparing fluorescent cellulose fiber (FCF). This method involved modification of cellulose fiber by sodium chloroacetate and self-assembly of cellulose and Eu3+ by coordination interaction. The fluorescence properties, crystallinity, element compositions and microscopic morphology of the fluorescent cellulose fiber were characterized. The preparation mechanism of fluorescent cellulose fiber was explained. The red was presented by the FCF when it was exposed under the ultraviolet at wavelength of 397 nm, which showed its potential application in the field of security. The self-assembled preparation method was expected to be a novel candidate for the preparation of cellulose-based fluorescent fibers.