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Fluorescence Presentation

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Angela Gi

on 21 January 2015

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Transcript of Fluorescence Presentation

Fluorescence
Basic Instrumentation
Procedure
Ground state molecule absorbs radiant energy (short wavelength, high energy)
Molecule goes into excited state
Molecule releases energy (long wavelength), returns to ground state
Fluorometry

Each type of molecule has characteristic excitation and fluorescence energies
Emission light (fluorescence) intensity is proportional to the concentration of the fluorophore
BACTEC MGIT 960 System
An automated instrument that utilizes sensors to measure
fluorescence
for rapid detection of
mycobacteria

growth
Specimen added to 7mls MGIT (Mycobacteria Growth Indicator Tube media)
Tubes contain fluorescent sensor at the bottom
Oxygen Fluorescence
Actively respiring micro organisms consume oxygen, cause tubes to fluoresce.
The system incubates and monitors these samples every 60 minutes.
Clinical Significance
Suitable for the detection of mycobacteria esp. Mycobacterium tuberculosis
Also used for drug susceptibility testing
Can process large volumes
Rapid detection
Conventional cultures for mycobacteria takes on average 4 weeks
Advantages of the System
Less expensive than other similar systems
ex: BACTEC 460TB system (radiometric)
Fully automated- safe to use
eliminates sharps
Fluorescence technology is highly sensitive
Able to hold up to 960 tubes which are monitored every hour.
BACTEC & TB
Tuberculosis continues to be a problem in developing countries.
M. tuberculosis = drug resistant*
Rapid identification is crucial for early treatment and prevention.
Summary
Fluorometry can be utilized by different kinds of systems.
molecules have different fluorescent properties
Not only utilized in Chemistry laboratories.
Specimens
BACTEC MGIT 980 System is able to run the tests with various specimen types such as:
Sputum
Pus
Gastric Aspirates
Bronchial Washings
Laryngeal Swabs

Basic Instrumentation Overview
Tissue
Urine
CSF
Synovial Fluid
Pleural Fluid

Source - emits excitation light
Attenuator - controls the light intensity
Primary filter (monochromater) - selects the wavelength best absorbed by the solution
Sample - emits radiant energy in all directions
Secondary filter (monochromater) - prevents incident light from striking the photodetector
Detector - at a right angle to the sample
http://www.finddiagnostics.org/export/sites/default/resource-centre/find_reports/pdfs/mgit_manual_nov_2007.pdf
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC85696/
http://www.annclinlabsci.org/content/31/3/279.full
http://www.finddiagnostics.org/export/sites/default/resource-centre/find_reports/pdfs/mgit_manual_nov_2007.pdf
http://www.powershow.com/view/3aee8e-MGE1N/BACTEC_MGIT_960_CULTURE_SYSTEM_FOR_MYCOBACTERIA_David_Lubasi_powerpoint_ppt_presentation
http://www.ipaqt.org/wp-content/uploads/2013/02/MGIT-product-brochure1.pdf
Bishop, Michael L., Edward P. Fody, and Larry E. Schoeff. "Fluorometry." Clinical Chemistry: Techniques, Principles, Correlations. Philadelphia: Wolters Kluwer Health/Lippincott Williams & Wilkins, 2010. N. pag. Print.
Bishop, Michael L., Edward P. Fody, and Larry E. Schoeff. "Fluorometry." Clinical Chemistry: Techniques, Principles, Correlations. Philadelphia: Wolters Kluwer Health/Lippincott Williams & Wilkins, 2010. N. pag. Print.
Guglielmo, Jeannie "Luminescence" Class Notes, 2014
The End.
Full transcript