Calcium Green

high-resolution terminology - matching measurements at high-resolution

Calcium Green

Description

Calcium Green denotes a family of extrinsic fluorophores applied for measurement of Ca²⁺ concentration.

Abbreviation: CaG

MitoPedia methods: Fluorometry 

Sources and applications

Calcium Green^TM is a registered trademark and available from Life Technologies (former Invitrogen) in several formulations.

All fluorophores of the Calcium Green family have the same fluorophoric moiety in their molecular structure and therefore near identical optical properties. They differ in their K_D values and are suitable for measuring different ranges of calcium concentrations.

• Calcium Green 1: K_D = 190 nM
• Calcium Green-2: K_D = 550 nM
• Calcium Green-5N: K_D = 14000 nM

Measurement

See Calcium for method-independent considerations related to measuring calcium levels, covering e.g.

• Considerations before starting a project involving calcium measurements;
• Ca²⁺ buffering;
• Calculation of free Ca²⁺ concentrations;
• Ca²⁺ calibration buffers.

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MitoPedia O2k and high-resolution respirometry: O2k-Open Support 

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Ca measurement

See O2k-Fluo LED2-Module for specific aspects of fluorescence measurements using the O2k.

On the fluorophores producers homepage detailed product information is available

• Long-Wavelength Calcium Indicators Product Information
• CalciumCalibration

General information on measuring calcium, e.g.

• Molecular Probes-The Handbook, Introduction to Ca2+ measurements with fluorescent indicators,
• Molecular Probes-The Handbook, Fluorescent Ca2+ indicators excited with visible light

The three Calcium Green fluorophores and Magnesium Green share the same fluorophore moiety in their molecular structure, therefore the optical properties are very similar and the same fluorescence sensor and filter set can be used. Since the used concentration of Calcium Green might vary according to the application, the optimum light intensity and amplification used may be different from those for Magnesium Green. However, the values recommended for Magnesium Green will provide a good starting point.

Signal and output

1. Signal: The O2k-Fluo LED2-Module is operated through the amperometric (Amp)-Channel of the O2k, with electric current (ampere [A]) as the primary signal.
2. Output: The focus of the output with Calcium Green is on Type A: Concentration, and Type C: Force, potential.

References

• Use of Calcium Green-5N with the O2k-Fluorescence LED2-Module

• Elustondo 2014 Biochim Biophys Acta

• Use of Calcium Green-5N with the other instruments in parallel with oxygen consumption measurements with the O2k.

• Anderson_2011_Am_J_Physiol_Heart_Circ_Physiol

Publications: CaG O2k-FluoRespirometer

	Year	Reference	Organism	Tissue;cell	Stress	Diseases
Deline 2021 Biochim Biophys Acta Bioenerg	2021	Deline ML, Grashei M, van Heijster FHA, Schilling F, Straub J, Fromme T (2021) Adenylate kinase derived ATP shapes respiration and calcium storage of isolated mitochondria. Biochim Biophys Acta Bioenerg 1862:148409.	Mouse	Liver
Duplaquet 2020 Elife	2020	Duplaquet L, Leroy C, Vinchent A, Paget S, Lefebvre J, Vanden Abeele F, Lancel S, Giffard F, Paumelle R, Bidaux G, Heliot L, Poulain L, Furlan A, Tulasne D (2020) Control of cell death/survival balance by the MET dependence receptor. Elife 9:e50041.	Human	HEK	Cell death
Spinazzi 2019 Proc Natl Acad Sci U S A	2019	Spinazzi M, Radaelli E, Horré K, Arranz AM, Gounko NV, Agostinis P, Maia TM, Impens F, Morais VA, Lopez-Lluch G, Serneels L, Navas P, De Strooper B (2019) PARL deficiency in mouse causes Complex III defects, coenzyme Q depletion, and Leigh-like syndrome. Proc Natl Acad Sci U S A 116:277-86. 10.1073/pnas.1811938116	Mouse	Nervous system		Neurodegenerative
Naszai 2019 Sci Rep	2019	Nászai A, Terhes E, Kaszaki J, Boros M, Juhász L (2019) Ca(2+)N it be measured? Detection of extramitochondrial calcium movement with high-resolution FluoRespirometry. Sci Rep 9:19229.	Rat	Liver
Campos 2017 Autophagy	2017	Campos JC, Queliconi BB, Bozi LHM, Bechara LRG, Dourado PMM, Andres AM, Jannig PR, Gomes KMS, Zambelli VO, Rocha-Resende C, Guatimosim S, Brum PC, Mochly-Rosen D, Gottlieb RA, Kowaltowski AJ, Ferreira JCB (2017) Exercise reestablishes autophagic flux and mitochondrial quality control in heart failure. Autophagy 13:1304-317.	Rat	Heart	Permeability transition	Cardiovascular
Liepinsh 2016 Biochem J	2016	Liepinsh E, Makrecka-Kuka M, Volska K, Kuka J, Makarova E, Antone U, Sevostjanovs E, Vilskersts R, Strods A, Tars K, Dambrova M (2016) Long-chain acylcarnitines determine ischaemia/reperfusion-induced damage in heart mitochondria. Biochem J 473:1191-202.	Rat	Heart	Ischemia-reperfusion
Elustondo 2014 Biochim Biophys Acta	2014	Elustondo PA, Negoda A, Kane CL, Kane DA, Pavlov EV (2014) Spermine selectively inhibits high-conductance, but not low-conductance calcium-induced permeability transition pore. Biochim Biophys Acta 1847:231–40.	Rat	Liver	Permeability transition

Abstracts: CaG O2k-FluoRespirometer

	Year	Reference	Organism	Tissue;cell	Stress	Diseases
Cecatto 2022 Abstract Bioblast	2022	5.1. «10+5» Cecatto Cristiane, Cardoso LHD, Gnaiger E (2022) Mitochondrial calcium uptake capacity is lower than calcium retention capacity in the presence and absence of cyclosporin A. Bioblast 2022: BEC Inaugural Conference. In: https://doi.org/10.26124/bec:2022-0001	Mouse	Liver
Nickel 2018 IOC130	2018	Defects in mitochondrial calcium uptake precede defects of the respiratory chain in X-linked Barth syndrome cardiomyopathy.	Mouse	Heart		Cardiovascular Inherited

O2k-Publications

• O2k-Publications: CaG O2k-FluoRespirometer
• O2k-Publications: Ca2+ O2k-ISE Module