Summary: | The measurement of cysteine in human urine and live cells is crucial for evaluating biological metabolism, monitoring and maintaining the immune system, preventing tissue/DNA damage caused by free radicals, preventing autoimmune diseases, and diagnosing disorders such as cystinuria and cancer. A method that uses a fluorescence turn-on probe and a portable fluorescence spectrometer device are crucial for highly sensitive, simple, rapid, and inexpensive cysteine detection. Herein, we present the synthesis and application of a benzimidazole-based fluorescent probe (<b>ABIA</b>) along with the design and development of a portable fluorescence spectrometer device (<b>CysDDev</b>) for detecting cysteine in simulated human urine. <b>ABIA</b> showed excellent selectivity and sensitivity in detecting cysteine over homocysteine, glutathione, and other amino acids with the response time of 1 min and demonstrated a detection limit of 16.3 nM using the developed <b>CysDDev</b>. Further, <b>ABIA</b> also demonstrated its utility in detecting intracellular cysteine, making it an excellent probe for bio-imaging assay.
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