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On‐line green fluorescent protein sensor with LED excitation
Author(s) -
RandersEichhorn Lisa,
Albano C. Renee,
Sipior Jeffrey,
Bentley William E.,
Rao Govind
Publication year - 1997
Publication title -
biotechnology and bioengineering
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.136
H-Index - 189
eISSN - 1097-0290
pISSN - 0006-3592
DOI - 10.1002/(sici)1097-0290(19970920)55:6<921::aid-bit9>3.0.co;2-i
Subject(s) - fluorescence , green fluorescent protein , materials science , photomultiplier , excitation , signal (programming language) , optical fiber , line (geometry) , light emitting diode , optics , optoelectronics , chemistry , detector , physics , biochemistry , geometry , mathematics , computer science , gene , programming language , quantum mechanics
We present an intensity based sensor designed for on‐line monitoring of green fluorescent protein, a revolutionary marker of protein expression. The device consisted of a blue light emitting diode as the excitation source. A band pass excitation filter cut off light longer than 490 nm. The light was directed into a bifurcated optical fiber bundle with the common end inserted into a stainless steel housing equipped with a quartz window. The fiber bundle and stainless steel housing are steam sterilizable. The emission radiation was collected through a long wave pass filter to reject the excitation light shorter than 505 nm and was detected by a photomultiplier tube. The signal was amplified and sent to a computer for recording time course data. The sensor was tested in an Escherichia coli fermentation of JM105 transformed with pBAD‐GFP. The on‐line signal was compared to off‐line fluorescence spectrophotometer measurements. The on‐line profile closely followed the off‐line. Western blot data showed that with a time shift, the sensor was able to both continuously and quantitatively monitor expression of green fluorescent protein on‐line in real time. © 1997 John Wiley & Sons, Inc. Biotechnol Bioeng 55 :921–926, 1997.