Intracellular oxygen mapping using a myoglobin-mCherry probe with fluorescence lifetime imaging

Rozhin Penjweini; Alessio Andreoni; Tilman Rosales; Jeonghan Kim; Michael D. Brenner; Dan L. Sackett; Jay H. Chung; Jay R. Knutson

doi:10.1117/1.JBO.23.10.107001

8 October 2018 Intracellular oxygen mapping using a myoglobin-mCherry probe with fluorescence lifetime imaging

Rozhin Penjweini, Alessio Andreoni, Tilman Rosales, Jeonghan Kim, Michael D. Brenner, Dan L. Sackett, Jay H. Chung, Jay R. Knutson

Author Affiliations +

Journal of Biomedical Optics, Vol. 23, Issue 10, 107001 (October 2018). https://doi.org/10.1117/1.JBO.23.10.107001

Abstract

Oxygen (O₂) is one of the most important biometabolites. In abundance, it serves as the limiting terminus of aerobic respiratory chains in the mitochondria of higher organisms; in deficit, it is a potent determinant of development and regulation of other physiological and therapeutic processes. Most knowledge on intracellular and interstitial concentration ([O₂]) is derived from mitochondria isolated from cells or tissue biopsies, providing detailed but nonnative insight into respiratory chain function. The possible loss of essential metabolites during isolation and disruption of the normal interactions of the organelle with the cytoskeleton may cause these data to misrepresent intact cells. Several optical methodologies were also developed, but they are often unable to detect heterogeneity of metabolic characteristics among different individual cells in the same culture, and most cannot detect heterogeneous consumption within different areas of a single cell. Here, we propose a noninvasive and highly sensitive fluorescence lifetime microscopy probe, myoglobin-mCherry, appropriate to intracellular targeting. Using our probe, we monitor mitochondrial contributions to O₂ consumption in A549 nonsmall cell lung cancer cells and we reveal heterogeneous [O₂] within the intracellular environments. The mitochondrial [O₂] at a single-cell level is also mapped by adding a peptide to target the probe to the mitochondria.

Citation Download Citation

Rozhin Penjweini, Alessio Andreoni, Tilman Rosales, Jeonghan Kim, Michael D. Brenner, Dan L. Sackett, Jay H. Chung, and Jay R. Knutson "Intracellular oxygen mapping using a myoglobin-mCherry probe with fluorescence lifetime imaging," Journal of Biomedical Optics 23(10), 107001 (8 October 2018). https://doi.org/10.1117/1.JBO.23.10.107001

Received: 3 July 2018; Accepted: 4 September 2018; Published: 8 October 2018

ACCESS THE FULL ARTICLE

JOURNAL ARTICLE
14 PAGES

DOWNLOAD PAPER SAVE TO MY LIBRARY