Nanometer scale resolution imaging of pH and reactive oxygen species on the outer membrane surface for fast and effective cell discrimination in cancer tissues.

2021 Fiscal Year Annual Research Report

• Project/Area Number: 19K05228
• Research Institution: Kanazawa University
• Principal Investigator: PUPPULIN LEONARDO 金沢大学, ナノ生命科学研究所, 特任助教 (40771580)
• Co-Investigator(Kenkyū-buntansha): 山田 忠明 京都府立医科大学, 医学(系)研究科(研究院), 准教授 (00507048) ; 細木 誠之 京都薬科大学, 薬学部, 准教授 (30433254)
• Project Period (FY): 2019-04-01 – 2022-03-31
• Keywords: cell surface pH / nanosensor / Raman spectroscopy / SERS

Outline of Annual Research Achievements

We continued to utilize our newly developed method for nano-sensor anchoring to the cell surface to measure and study the pH of single cells. We analyzed new cell lines to quantify surface pH, including A549 lung cancer cells and human keratinocytes (HKC). Based on these results, we prepared and submitted an invited paper (分光便利帳) to the Journal of the Spectroscopical Society of Japan(分光研究).
In addition, we focused our efforts on developing the anchoring of the gold nanosensor for pH measurement on fully confluent cell sample, mimicking the condition for analysis of cell tissues. We optimized the protocol using higher concentration of gold nanoparticles and laser line illumination mode for simultaneous acquisition of 400 SERS spectra from a wide area of the sample. We analyzed different cell lines: pancreatic cancer cell BxPC; pancreatic cancer cells AsPC-1; murine fibroblast cells NIH/3T3; human fibroblast WI-38; lung cancer A549. Based on the collected results, we are currently preparing a manuscript for publication on a scientific journal.