Background Splenectomy is an effective intervention in primary immune thrombocytopenia (ITP). Attempts to define pre-clinical predictors of platelet response to splenectomy are inconsistent. Based on international studies defining the likelihood of platelet response using platelet sequestration, patients with relapsed/refractory ITP being considered for splenectomy at a regional Australian hospital were assessed with 111 Indium-labelled autologous platelet sequestration (ILAPS) studies. Aims To audit the use of ILAPS in an Australian setting and define its role in predicting response to splenectomy. Methods A retrospective review of all patients referred for an ILAPS study at a regional hospital was performed. Results for each patient were expressed as an "R" value (spleen/ liver uptake ratio) to quantify the platelet sequestration pattern and outcome post splenectomy, based on platelet counts. Results A total of 45 patients were identified, 13 underwent splenectomy and 32 were medically managed. Patients with favourable ILAPS scans (pure or predominant splenic sequestration) demonstrated a superior response post splenectomy (100% overall response rate (ORR), 83.5% complete remission (CR)) compared to those with unfavourable ILAPS scans (mixed or pure hepatic sequestration) (71.4% ORR, 57.1% CR) over 12 months. Conclusions The use of ILAPS in the Australian setting is feasible and this experience confirms larger international studies demonstrating its utility as a predictor of response to splenectomy in ITP. An unfavourable ILAPS scan could be considered a negative predictor of response prompting consideration for other emerging ITP treatments such as thrombopoietin-receptor agonists (TPO-RAs) or B-cell depleting therapy such as Rituximab. This article is protected by copyright. All rights reserved.