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In this paper, we demonstrate the possibility of using spotlights for 5D guidance of a microsurgical instrument. The theoretical basis of the localization for the instrument based on the projection of a single spotlight is analyzed to deduce the position and orientation of the spotlight source. The usage of multiple spotlights is also proposed to check the possibility of further improvements for the performance boundaries. The proposed method is verified within a high-fidelity simulation environment using the 3D creation suite Blender. Experimental results show that the average positioning error is 0.029 mm using a single spotlight and 0.025 mm with three spotlights, respectively, while the rotational errors are 0.124 and 0.101, which shows the application to be promising in instrument localization for retinal surgery.<\/jats:p>","DOI":"10.1017\/s0263574722001862","type":"journal-article","created":{"date-parts":[[2023,1,26]],"date-time":"2023-01-26T11:04:16Z","timestamp":1674731056000},"page":"1536-1549","source":"Crossref","is-referenced-by-count":6,"title":["Theoretical error analysis of spotlight-based instrument localization for retinal surgery"],"prefix":"10.1017","volume":"41","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-6944-1483","authenticated-orcid":false,"given":"Mingchuan","family":"Zhou","sequence":"first","affiliation":[]},{"given":"Felix","family":"Hennerkes","sequence":"additional","affiliation":[]},{"given":"Jingsong","family":"Liu","sequence":"additional","affiliation":[]},{"given":"Zhongliang","family":"Jiang","sequence":"additional","affiliation":[]},{"given":"Thomas","family":"Wendler","sequence":"additional","affiliation":[]},{"given":"M. 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