Whole-animal imaging of bacterial infection using endoscopic excitation of β-lactamase (BlaC)-specific fluorogenic probe
Autor: | Hee-Jeong Yang, Yunfeng Cheng, Jianghong Rao, Fatemeh Nooshabadi, Kristen C. Maitland, Jeffrey D. Cirillo, Hexin Xie |
---|---|
Rok vydání: | 2016 |
Předmět: |
0301 basic medicine
Colony-forming unit Fluorescence-lifetime imaging microscopy Tuberculosis biology Chemistry Substrate (chemistry) Nanotechnology medicine.disease biology.organism_classification Fluorescence law.invention Microbiology Mycobacterium tuberculosis 03 medical and health sciences 030104 developmental biology law medicine Recombinant DNA Bacteria |
Zdroj: | SPIE Proceedings. |
ISSN: | 0277-786X |
DOI: | 10.1117/12.2213820 |
Popis: | Tuberculosis (TB), caused by Mycobacterium tuberculosis (Mtb), remains one of the most frequent causes of death worldwide. The slow growth rate of Mtb limits progress toward understanding tuberculosis including diagnosis of infections and evaluating therapeutic efficacy. Development of near-infrared (NIR) β-lactamase (BlaC)-specific fluorogenic substrate has made a significant breakthrough in the whole-animal imaging to detect Mtb infection. The reporter enzyme fluorescence (REF) system using a BlaC-specific fluorogenic substrate has improved the detection sensitivity in whole-animal optical imaging down to ~104 colony forming units (CFU) of bacteria, about 100-fold improvement over recombinant strains. However, improvement of detection sensitivity is strongly needed for clinical diagnosis of early stage infection at greater tissue depth. In order to improve detection sensitivity, we have integrated a fiber-based microendoscpe into a whole-animal imaging system to transmit the excitation light from the fiber bundle to the fluorescent target directly and measure fluorescent level using BlaC-specific REF substrate in the mouse lung. REF substrate, CNIR800, was delivered via aerosol route to the pulmonary infected mice with M. bovis BCG strain at 24 hours post-infection and groups of mice were imaged at 1–4 hours post-administration of the substrate using the integrated imaging system. In this study we evaluated the kinetics of CNIR800 substrate using REF technology using the integrated imaging system. Integration of these technologies has great promise for improved detection sensitivity allowing pre-clinical imaging for evaluation of new therapeutic agents. |
Databáze: | OpenAIRE |
Externí odkaz: |