Yeast do not seem to form biofilms on the bottoms of corks when they’re used, rather than metal crown caps, to secure Champagne bottles during their in-bottle secondary fermentation. This, at least, is the conclusion of an article in the current issue of the American Journal of Enology and Viticulture (paywall), in which Burgundy-based authors investigated the question for the sake of understanding whether Champagne producers, some of whom are using cork for their longer-aging wines, risked upsetting in-bottle fermentation dynamics. After a year of bottle fermentation and aging, a few cells apparently got caught in porous crevices of the cork, but systematic growth presaging the tenacity of a biofilm wasn’t happening.
That finding will no doubt interest the odd sparkling wine producer. Much more interesting is the method they used to make “invisible” cells visible and so reach that conclusion.
How do you determine whether a microbial biofilm is growing somewhere?
Before it reaches scrape-it-off-with-a-fingernail thickness, the answer is usually microscopy. But what kind of microscopy? Let’s say you want to see the very earliest stages of what might become a biofilm. You want to visualize individual yeast cells sticking to a rough, craggy surface with lots of crevices for hiding—the cork. Pointing a light microscope at the cork won’t do. The kind that you probably used in school to see cells suspended in liquid (from your cheek, or bacteria from your teeth), and that many small wine and brewery labs use to check for live cells requires that whatever you’re looking at be thin enough for light to pass through. Slicing corks into sections thin enough for light microscopy might destroy or displace any would-be-biofilm-forming yeast cells. Moreover, can you imagine scanning the bottom of cork after cork, continually asking yourself: is that a yeast cell, or a bit of cork shaped like a yeast cell? A digital image analysis program trained to recognize yeast cells might address the latter problem (with some degree of error), but the whole task clearly calls for a more sophisticated technique.
When your sample isn’t transparent enough for light microscopy, fluorescence microscopy can be an alternative; instead of visualizing light passing through the sample, fluorescence microscopy relies on whateveritis you’re examining absorbing and then emitting light—fluorescing—which can then be picked up by the microscope’s detector.* Conveniently, cork and other plant matter comes with a built-in fluorescent molecule; lignin, a rigid polymer and major contributor to the stiffness of wood and bark, is easy to see under the microscope …