In situ formation of a gel microbead for indirect laser micromanipulation of microorganisms

Abstract

We propose the in situ formation of gel microbeads made of a thermoreversible hydrogel for indirect laser micromanipulation of microorganisms. Irradiation, using a 1064nm laser, of an aqueous solution mixed with poly-(N-isopropylacrylamide) through a high magnification lens resulted in the formation of a gel microbead at the laser focus due to heating. The gel microbead is trapped by the laser, and is used for indirect laser micromanipulation of microorganisms. However, the laser power used to form the bead is generally too strong to perform manipulation in a stable manner. In this letter we show a method to reduce the laser power to form a gel microbead using the poly-(N-isopropylacrylamide) aqueous solution by the addition of additives. The gelation temperature and the laser absorption rate of the solution in the presence of several different additives were investigated. We selected YPD (yeast extract, peptone, dextrose) broth as an additive and measured the relationship between the laser power, irradiation time, and diameter of the gel microbead. We succeeded in reducing the laser power for gel microbead formation, and in using the laser-trapped gel microbead for the manipulation of a yeast cell and DNA.