Combined imaging of bacteria and oxygen in biofilms.
Transparent sensors for microscopic O(2) imaging were developed by spin coating an ultrathin (<1- to 2-microm) layer of a luminescent O(2) indicator onto coverslips. The sensors showed (i) an ideal Stern-Volmer quenching behavior of the luminescence lifetime towards O(2) levels, (ii) homogeneous measuring characteristics over the sensor surface, and (iii) a linear decline of luminescence lifetime with increasing temperature. When a batch of such coverslip sensors has been characterized, their use is thus essentially calibration free at a known temperature. The sensors are easy to use in flow chambers and other growth devices used in microbiology. We present the first application for combined imaging of O(2) and bacteria in a biofilm flow chamber mounted on a microscope equipped with a spinning-disk confocal unit and a luminescence lifetime camera system.
Kühl
Michael
M
Marine Biological Laboratory, Department of Biology, University of Copenhagen, Strandpromenaden 5, DK-3000 Helsingør, Denmark. mkuhl@bi.ku.dk
Rickelt
Lars F
LF
Thar
Roland
R
D001419:Bacteria / Q000254:growth & development* / Q000378:metabolism; D018441:Biofilms / Q000254:growth & development*; D015374:Biosensing Techniques / Q000295:instrumentation*; D004581:Electronics; D008163:Luminescent Measurements / Q000295:instrumentation* / Q000379:methods; D018613:Microscopy, Confocal; D010100:Oxygen / Q000032:analysis* / Q000737:chemistry
D010100:Oxygen
Appl Environ Microbiol. 1999 Sep;65(9):4108-17
10473423
Methods Enzymol. 1999;310:20-42
10547780
Methods Enzymol. 1999;310:160-6
10547790
Anal Chem. 1999 Dec 1;71(23):793A-800A
10596206
Methods Enzymol. 2004;381:715-35
15063708
Methods Enzymol. 2005;397:147-66
16260290
Methods Enzymol. 2005;397:166-99
16260291
J Biomed Opt. 2006 May-Jun;11(3):34001
16822051
Biophys J. 1991 Dec;60(6):1374-87
1723311
Nature. 2007 Feb 1;445(7127):533-6
17268468
J Exp Biol. 2007 Mar;210(Pt 6):1006-14
17337713
J Microbiol Methods. 2007 Aug;70(2):336-45
17590463
Biotechnol Bioeng. 1994 May;43(11):1131-8
18615526
J Phycol. 2008 Jun;44(3):541-50
27041414