Dual wavelength fluorescent ratiometric pH measurement by scanning near-field optical microscopy

Yongbo Li; Ryosuke Shinohara; Kentaro Iwami; Yoshihiro Ohta; Norihiro Umeda

doi:10.1117/12.885194

28 December 2010 Dual wavelength fluorescent ratiometric pH measurement by scanning near-field optical microscopy

Yongbo Li, Ryosuke Shinohara, Kentaro Iwami, Yoshihiro Ohta, Norihiro Umeda

Proceedings Volume 7544, Sixth International Symposium on Precision Engineering Measurements and Instrumentation; 754419 (2010) https://doi.org/10.1117/12.885194
Event: Sixth International Symposium on Precision Engineering Measurements and Instrumentation, 2010, Hangzhou, China

Abstract

A novel method to observe pH distribution by dual wavelength fluorescent ratiometric pH measurement by scanning near-field optical microscopy (SNOM) is developed. In this method, in order to investigate not only the pH of mitochondrial membrane but also its distribution in the vicinity, a pH sensitive fluorescent reagent covers mitochondria instead of injecting it to mitochondria. This method utilizes a dual-emission pH sensitive dye and SNOM with a themally-pulled and metal-coated optical fiber to improve the spatial resolution. Time-dependence of Fluorescent intensity ratio (FIR) under acid addition is investigated. As the distances between the dropped point and the SNOM probe becomes closer, the time when FIR changes becomes earlier. The response of mitochondria under supplement of nutrition is studied by using this method. While the probe is near to mitochondria, the ratio quickly becomes to increase. In conclusion, it was confirmed that the temporal variation of pH can be detected by this method, and pH distribution in the vicinity of mitochondria is able to be measured by this method.

Citation Download Citation

Yongbo Li, Ryosuke Shinohara, Kentaro Iwami, Yoshihiro Ohta, and Norihiro Umeda "Dual wavelength fluorescent ratiometric pH measurement by scanning near-field optical microscopy", Proc. SPIE 7544, Sixth International Symposium on Precision Engineering Measurements and Instrumentation, 754419 (28 December 2010); https://doi.org/10.1117/12.885194

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