Purpose :Triple negative breast cancers (TNBCs) have a high mortality rate owing to aggressive proliferation and metastasis and a lack of effective therapeutic target. Therefore, an urgent and unmet need remains to identify new and effective molecular targets and associated therapies in TNBC. Recently, intercellular adhesion molecule-1 (ICAM-1) have been demonstrated that is highly overexpressed in human TNBC tissues and cell lines. We present novel ICAM-1 -specific probes for in vivo evaluation of the receptor status by near-infrared (NIR) optical imaging. ICAM-1 would lead to the development of a new strategy and platform for TNBC therapy and diagnosis. Methods : Affibody molecules peptides γ3 were synthesized and labeled with FITC ,NIR-fluorescent dyes cy5.5 . The binding affinity and specificity of the obtained probe were tested in vitro. For in vivo validation, the relationship of the measured NIR signal and ICAM-1 expression was characterized in two breast cancer xenograft models, expressing different levels of ICAM-1. Accumulation of Affibody molecules in tumor tissue was further confirmed by ex vivo analysis. Results: γ3-FITC or γ3-cy5.5 conjugates showed high affinity to ICAM-1 . These formulations are no cytotoxicity in TNBC cell line MDA-MB-231 and NON-TNBC cell line MCF-7. Fluorescence accumulation in MDA-MB-231 xenografts was evident as soon as a few minutes post injection and slowly reached its maximum. On the other hand, just a few signal retention was observed in MCF-7 xenografts. Immunostaining of extracted tumor tissue confirmed penetration of the tracer into tumor tissue. Conclusions: we have successfully engineered a novel TNBC-targeting Affibody molecules probe. Synthesized ICAM-1-targeted probe is a powerful tool to monitor ICAM-1 status in a preclinical setting. The design introduced in our study provides a new platform for future TNBC-targeted diagnosis and therapeutics.