We report a comparative study of the efficacy of immobilizing living bacteria by means of seven antibodies against bacterial surface antigens associated with Salmonella enterica Serovar Typhimurium. The targeted bacterial antigens were CFA/I fimbriae, flagella, lipopolysaccharides (LPS), and capsular F1 antigen. The best immobilization of S. Typhimurium was achieved with the antibody against CFA/I fimbriae. The immobilization of bacteria using antiflagellin showed significant enhancement if the flagella rotary motion was paralyzed. Of the four antibodies targeting LPS structures, only one, the antibody against the O-antigen polysaccharides, showed a relatively efficient bacterial immobilization. No bacterial immobilization was achieved using the antibody against F1 antigen, presumably because F1 protein can detach from the bacterial surface easily. The results suggest that an antibody for bacterial immunoimmobilization should target a surface antigen which extends out from the bacterial surface and is tightly attached to the bacterial cell wall. The microarrays of living S. Typhimurium cells immobilized in this manner remained viable and effective for at least 2 weeks in growth medium before a thick biofilm covered the whole surface.