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Although several technologies have been developed to isolate cells of interest from a heterogenous sample, clogging and impaired cell viability limit such isolation. We have developed the Enrich TROVO system as a novel, nonfluidic technology to sort live cells. The TROVO system combines imaging-based cell selection and photo-crosslinking of (gelatin methacrylate) gelMA-hydrogel to capture cells. After capture, cells are released by enzymatic digestion of the hydrogel and then retrieved for downstream analysis or further cell culturing. The system can capture cells with a recovery rate of 48% while maintaining 90% viability. Moreover, TROVO can enrich rare cells 506-fold with 93% efficiency using single step isolation from a 1:10 4 cell mixture, and can also capture one target cell from 1 million cells, reaching an enrichment ratio of 9128. In addition, 100% purity and 49% recovery rate can be achieved by a following negative isolation process. Compared to existing technologies, the TROVO system is clog-resistant, highly biocompatible, and can process a wide range of sample sizes., Competing Interests: Declaration of Competing Interest The authors of this manuscript declare that they have no competing financial, professional, or personal interests that could have influenced the work reported in this paper. The research conducted and presented in this manuscript was carried out independently and without any external influence. This research was funded by NIAID R44AI147734, but the funding agency had no role in the study design, data collection, analysis, interpretation, or writing of this manuscript. We confirm that the manuscript has been read and approved by all named authors, and there are no other persons who satisfied the criteria for authorship but are not listed. We further confirm that the order of authors listed in the manuscript has been approved by all of us. We understand that any undisclosed competing interests may result in the retraction of the manuscript, and we agree to promptly inform the journal editor if any potential competing interests arise in the future. Please let us know if you require any further information or clarification regarding our declaration of interest statement., (Copyright © 2024 The Authors. Published by Elsevier B.V. All rights reserved.)