Microscopy methods to investigate structure of potable water biofilms

Walker, James T., Verran, Joanna, Boyd, Robert D. and Percival, Steven (2001) Microscopy methods to investigate structure of potable water biofilms. In: Microbial Growth in Biofilms - Part B: special environments and physicochemical aspects. Methods in Enzymology (337). Academic Press, pp. 243-255. ISBN 9780121822385

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Abstract

This chapter discusses a number of methods used for the visualization of biofilm that has been developed primarily on plumbing tube materials. Differential interference contrast (DIC) and fuorescence microscopy examination was carded out using a Nikon Labophot-2 that combined both epifluorescence and DIC. Photography was carded out using a Nikon F-801 35 mm camera and a JVC TK-1085E color video camera head with built-in compact power source fitted to a trinocular head. Using the video camera, visualization of the subject matter was relayed either to a video monitor for time lapse photography or to the multisync monitor (Taxan) for the image analysis program—both of which increased magnification to greater than 1500 times. A Medical Research Council (MRC)-600 (Bio-pad) fitted with an argon laser with maximum emission at 488 nm and excitation at 514 was mounted in the upright position above an optical light microscope. The images from two different fluorescent markers can be imaged simultaneously using an excitation filter and dichroic mirror to direct the image to either photomultiplier l or 2. These filters could be changed without disrupting the optical alignment of the microscope, thus allowing imaging of a single field of view under different spectral conditions.