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Abstract #1846

Real-time 13C J-coupling-edited Proton High-Resolution Magic Angle Spinning (HRMAS) MRS of Live Cells

Rajshree Ghosh Biswas1, Ella Zhang1, Aidan Pavao2, Lynn Bry2, and Leo Cheng3
1Radiology, Massachusetts General Hospital, Charlestown, MA, United States, 2Pathology, Brigham and Women's Hospital, Boston, MA, United States, 3Harvard T.H. Chan School of Public Health, Boston, MA, United States

Synopsis

Keywords: Data Acquisition, Acquisition Methods, In vivo, microbial MRS, 1H[13C-Jed], HRMAS

Motivation: Real-time biochemical changes at the cellular-level can be monitored using 13C-labelled substrates via 13C-MRS. However, a small sample volume, spectral overlap and low sensitivity of MRS make long-term monitoring of targeted metabolic pathways challenging.

Goal(s): Use 1H-13C-J-coupling (1H[13C-Jed]) to observe protons attached to 13C-labelled substrates, enabling targeted metabolic process monitoring, in vivo, with increased sensitivity.

Approach: Pathogenic Clostridioides difficile was grown in 13C-media (13C-glucose and 13C-threonine). 13C-HRMAS MRS of 100, 000 cells (20 µL), was used to monitor cellular metabolism for >48 hours.

Results: Downstream metabolic breakdown products and a 13.9X improvement in S/N by 1H[13C-Jed], compared to standard 13C-MRS, was observed.

Impact: 1H-13C-J-coupling (1H[13C-Jed]) HRMAS was used to follow real-time targeted metabolism of 13C-glucose and 13C-threonine in 100,000 C. diff. cells (20 µL) using 13C-labelled substrates, for >48 hours. A 13.9x improvement in S/N in 1H[13C-Jed relative to 13C-MRS was observed.

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