回折限界を超える空間分解能で生細胞内の酸素濃度情報を可視化する手法の確立を目的とした．ガウシアンビームを分岐して一方で中空ビームを生成し，それぞれで対象を2次元走査し，得られた画像の差分を取ることで超解像を得た．まず，差分過程における輝度情報損失が少なく，高い空間分解能を得るための光学条件を決定し，実測実験からおよそ100 nmの分解能を達成した．さらに酸素分圧の定量に必要なリン光寿命を高速決定するアルゴリズムを適用した．以上より，提案法は簡便かつ高速に生細胞の超解像蛍光画像を得る手法として有効であることを示した．
The establishment of a method to visualize oxygen concentration information in living cells with spatial resolution beyond the diffraction limit will contribute to the elucidation of local life phenomena. A laser beam was split to generate a Gaussian beam and a doughnut-shaped beam. The object was scanned two-dimensionally with each beam, and super-resolution images were obtained by subtracting the obtained images. Optical conditions were determined to minimize the loss of intensity information during the differencing process and to obtain high spatial resolution. As a result, a resolution of approximately 100 nm was achieved based on experimental measurements. Furthermore, an algorithm to rapidly determine the phosphorescence lifetime for quantifying the oxygen partial pressure enables fast imaging. Taken together, the proposed method is expected to be used for fast super-resolution oxygen partial pressure imaging of living cells.

研究種目 : 基盤研究 (C) (一般)
研究期間 : 2019～2021
課題番号 : 19K12762
研究分野 : 生体医工学