Deposition of Bacteria from Sessile Drops

Abstract

This dissertation reports on the discovery of a new method of patterning bacteria (Pseudomonas aeruginosa PAO1) on a surface using a drying sessile drop. This work identifies bacterial suspension age and the length of time mica is exposed to the laboratory atmosphere as the key parameters which impact the behavior of the sessile drop and the resulting residue. Possible origins of mica aging and bacterial suspension aging are discussed in light of the literature and the experimental conditions. The residue area and the fraction of the residue area on which substantial bacteria and salt deposits remained after the drying of the drop (fill-in fraction) were measured via analysis of optical micrographs. In general, smaller residues are more filled in. For fresh bacterial suspensions, and short mica exposure times, the residue covers the largest area and is characterized by rings formed during discrete depinning events as the solvent evaporates. As the exposure time increases and the mica surface slowly picks up contaminants from the atmosphere, the drop residue shrinks in size and bacteria are deposited in a regular cellular film in the interior of the drop residue. The fraction of the interior area covered by the cellular film is well correlated with the mica exposure time. For sufficiently aged bacterial suspensions, residues are small and more filled-in than residues formed from fresh suspensions on similarly aged mica. In addition, the interior deposition pattern transitions from a cellular film characteristic of fresh suspensions to a cracked carpet pattern for aged suspensions. Suspension aging related changes in the residues are attributed to accumulation of organic materials such as DNA, RNA, proteins, and other bacterial components in the suspension. The suspension aging process is also observed to be at least partially dependent on ventilation of the suspension during aging.