SEM imaging of the stimulatory response of RAW264.7 cells against Porphyromonas gingivalis using a simple technique employing new conductive materials.

Abstract

In the medical biology, it is essential to understand not only biological morphology but also the interaction between biological materials and agents. To study these, electron microscopy (EM) is often utilized. However, sample preparation techniques for EM require a high level of skill and a considerable time. Here, we conducted EM using a simple technique employing a conductive liquid, BEL-1, and compared the results with another simple technique employing an ionic liquid, 1-butyl-3-methylimidazolium tetrafluoroborate ([BMIM][BF₄ ]). BEL-1 was used for sample pretreatment, and the morphologies of the mouse RAW 264.7 cell line, Porphyromonas gingivalis, and the RAW 264.7 cell line were stimulated via co-incubation with P. gingivalis and observed using field emission scanning EM (FE-SEM). In the present study, the inflammation-induced system of P. gingivalis was successfully established. FE-SEM results revealed the fine morphology of the RAW 264.7 cell line and P. gingivalis and confirmed a morphological change in the RAW 264.7 cell line caused by P. gingivalis stimulation. Using the developed sample preparation technique employing BEL-1, high-contrast and high-resolution observations of deformable biological materials were conducted without any difficulty or the necessity for complicated technique. This morphological information and the developed techniques can contribute to reveal the interaction between biological materials and agents and thereby accelerate drug formulation and disease treatment. © 2017 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater, 106B: 1280-1285, 2018.