Oxygen sensing PLIM together with FLIM of intrinsic cellular fluorophores for metabolic mapping

Sviatlana Kalinina; Patrick Schaefer; Jasmin Breymayer; Dominik Bisinger; Sabyasachi Chakrabortty; Angelika Rueck

doi:10.1117/12.2287429

20 February 2018 Oxygen sensing PLIM together with FLIM of intrinsic cellular fluorophores for metabolic mapping

Sviatlana Kalinina, Patrick Schaefer, Jasmin Breymayer, Dominik Bisinger, Sabyasachi Chakrabortty, Angelika Rueck

Proceedings Volume 10497, Imaging, Manipulation, and Analysis of Biomolecules, Cells, and Tissues XVI; 104970F (2018) https://doi.org/10.1117/12.2287429
Event: SPIE BiOS, 2018, San Francisco, California, United States

Abstract

Otical imaging techniques based on time correlated single photon counting (TCSPC) has found wide applications in medicine and biology. Non-invasive and information-rich fluorescence lifetime imaging microscopy (FLIM) is successfully used for monitoring fluorescent intrinsic metabolic coenzymes as NAD(P)H (nicotinamide adenine dinucleotide (phosphate)) and FAD+ (flavin adenine dinucleotide) in living cells and tissues. The ratio between proteinbound and free coenzymes gives an information about the balance between oxidative phosphorylation and glycolysis in the cells. The changes of the ratio reflects major cellular disorders. A correlation exists between metabolic activity, redox ratio and fluorescence lifetime during stem cell differentiation, neurodegenerative diseases, and carcinogenesis.

A multichannel FLIM detection system was designed for monitoring the redox state of NAD(P)H and FAD+ and other intrinsic fluorophores as protoporphyrin IX. In addition, the unique upgrade is useful to perform FLIM and PLIM (phosphorescence lifetime imaging microscopy) simultaneously. PLIM is a promising method to investigate oxygen sensing in biomedical samples. In detail, the oxygen-dependent quenching of phosphorescence of some compounds as transition metal complexes enables measuring of oxygen partial pressure (pO₂). Using a two-channel FLIM/PLIM system we monitored intrinsic pO₂ by PLIM simultaneously with NAD(P)H by FLIM providing complex metabolic and redox imaging of living cells.

Physico-chemical properties of oxygen sensitive probes define certain parameters including their localisation. We present results of some ruthenium based complexes including those specifically bound to mitochondria.

Conference Presentation

Citation Download Citation

Sviatlana Kalinina, Patrick Schaefer, Jasmin Breymayer, Dominik Bisinger, Sabyasachi Chakrabortty, and Angelika Rueck "Oxygen sensing PLIM together with FLIM of intrinsic cellular fluorophores for metabolic mapping ", Proc. SPIE 10497, Imaging, Manipulation, and Analysis of Biomolecules, Cells, and Tissues XVI, 104970F (20 February 2018); https://doi.org/10.1117/12.2287429

ACCESS THE FULL ARTICLE

INSTITUTIONAL
Select your institution to access the SPIE Digital Library.

SELECT YOUR INSTITUTION

PERSONAL
Sign in with your SPIE account to access your personal subscriptions or to use specific features such as save to my library, sign up for alerts, save searches, etc.

PERSONAL SIGN IN

No SPIE Account? Create one

PURCHASE THIS CONTENT

SUBSCRIBE TO DIGITAL LIBRARY

50 downloads per 1-year subscription

Members: $195

Non-members: $335 ADD TO CART

25 downloads per 1 - year subscription

Members: $145

Non-members: $250 ADD TO CART

PURCHASE SINGLE ARTICLE

Includes PDF, HTML & Video, when available