Sentinel lymph node is the first regional lymph node that drains the lymph from the primary tumor. It is potentially the first node to receive the seeding of lymph-borne metastatic cells. This study aimed to discuss lymphoscintigraphy procedural guidelines for detection of sentinel node using 99mTc-Phytate in Isfahan, Iran. Moreover, the preliminary results of the first year's clinical experience of lymphoscintigraphy in Isfahan, Iran are also presented. A total of 36 consecutive sentinel node procedures were performed following our protocol in March 2013 to March 2014. For all 36 patients, after intradermal injection of 0.5–1 mCi of 99mTc-Phytate, 5, 30 and 120 min with hands up lymphoscintigraphy was performed. All procedures were performed in a 1-day setting with 99mTc-Phytate injection in intradermal volume of about 0.1 cc. At 5, 30 and 120 min after injection, anterior and lateral images (4 min), were acquired using gamma-camera (energy 140 keV, window 15–20% and LEHR collimator). For all patients, at least one axillary sentinel lymph node was detected. For three patients, 2 SNs were seen. The images 5 min after injection showed at least one axillary sentinel node in 18 of 36 patients. However for the remaining patients, more delayed images (after 30 and 120 min) were needed. Although, no changes were seen in 120 min images compared to 30 min images. Considering the used protocol, from the evaluated data it can be concluded that lymphoscintigraphy after 30 min periareolar injection of about 0.5–1 mCi 99mTc-Phytate in an intradermal volume of about 0.1 cc yields an axillary sentinel node in all the patients. Imaging 120 min after injection is of no additional value and can be omitted.