Calcium Green: Difference between revisions
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=== Ca measurement === | === Ca measurement === | ||
:::: | :::: In high-resolution respirometry the CaG is used with the [[O2k-FluoRespirometer]] with [O2k-Fluo Smart-Module]] or [[O2k-Fluo LED2-Module]] selecting the [[Fluorescence-Sensor Blue]]/[[Filter Set MgG / CaG]]. Please see these pages for specific aspects of fluorescence measurements using the O2k. | ||
:::: {{Template:O2k-Fluorometry: black stoppers}} | |||
:::: {{Template:O2k-Fluorometry: switch off chambers illumination}} | |||
:::: 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 Blue|fluorescence sensor]] and [[Filter Set MgG / CaG|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 [[Magnesium_Green#Instrument|values recommended for Magnesium Green]] will provide a good starting point. | |||
Β | |||
:::: On the fluorophores producers homepage detailed product information is available Β | :::: On the fluorophores producers homepage detailed product information is available Β | ||
::::* [https://tools.lifetechnologies.com/content/sfs/manuals/mp03010.pdf Long-Wavelength Calcium Indicators Product Information ] | ::::* [https://tools.lifetechnologies.com/content/sfs/manuals/mp03010.pdf Long-Wavelength Calcium Indicators Product Information ] | ||
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::::* [http://www.lifetechnologies.com/at/en/home/references/molecular-probes-the-handbook/indicators-for-ca2-mg2-zn2-and-other-metal-ions/introduction-to-ca2-measurements-with-fluorescent-indicators.html Molecular Probes-The Handbook, Introduction to Ca2+ measurements with fluorescent indicators], | ::::* [http://www.lifetechnologies.com/at/en/home/references/molecular-probes-the-handbook/indicators-for-ca2-mg2-zn2-and-other-metal-ions/introduction-to-ca2-measurements-with-fluorescent-indicators.html Molecular Probes-The Handbook, Introduction to Ca2+ measurements with fluorescent indicators], | ||
::::* [http://www.lifetechnologies.com/at/en/home/references/molecular-probes-the-handbook/indicators-for-ca2-mg2-zn2-and-other-metal-ions/fluorescent-ca2-indicators-excited-with-visible-light.html Molecular Probes-The Handbook, Fluorescent Ca2+ indicators excited with visible light] | ::::* [http://www.lifetechnologies.com/at/en/home/references/molecular-probes-the-handbook/indicators-for-ca2-mg2-zn2-and-other-metal-ions/fluorescent-ca2-indicators-excited-with-visible-light.html Molecular Probes-The Handbook, Fluorescent Ca2+ indicators excited with visible light] | ||
Revision as of 12:27, 19 March 2020
Description
Calcium Green denotes a family of extrinsic fluorophores applied for measurement of Ca2+ concentration.
Abbreviation: CaG
MitoPedia methods:
Fluorometry
Sources and applications
- Calcium GreenTM 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 KD values and are suitable for measuring different ranges of calcium concentrations.
- Calcium Green 1: KD = 190 nM
- Calcium Green-2: KD = 550 nM
- Calcium Green-5N: KD = 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.
- See Calcium for method-independent considerations related to measuring calcium levels, covering e.g.
- Measurement
- Colour of media and chemicals used
- For all fluorometric techniques, special care must be taken to avoid colored chemicals that might affect the fluorescence signal (excitation and emission wavelength spectra).
- With CaG fluorophore, some uncouplers that present a yellow color might affect the signal.
- Colour of media and chemicals used
MitoPedia O2k and high-resolution respirometry:
O2k-Open Support
Ca measurement
- In high-resolution respirometry the CaG is used with the O2k-FluoRespirometer with [O2k-Fluo Smart-Module]] or O2k-Fluo LED2-Module selecting the Fluorescence-Sensor Blue/Filter Set MgG / CaG. Please see these pages for specific aspects of fluorescence measurements using the O2k.
- Use black stoppers with black cover-slips to exclude disturbances by external light sources.
- Before the experiments, switch off the illumination of the chambers (in [Oroboros O2k] \ [ O2k control ], or [F7]).
- 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.
- On the fluorophores producers homepage detailed product information is available
- General information on measuring calcium, e.g.
Signal and output
- 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.
- 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
- Use of Calcium Green-5N with the other instruments in parallel with oxygen consumption measurements with the O2k.
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, HorrΓ© 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 | NΓ‘szai A, Terhes E, Kaszaki J, Boros M, JuhΓ‘sz 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 1847:231β40. | Rat | Liver | Permeability transition |
- Abstracts: CaG O2k-FluoRespirometer
Year | Reference | Organism | Tissue;cell | Stress | Diseases | |
---|---|---|---|---|---|---|
Cecatto 2022 Abstract Bioblast | 2022 | 5.1. Β«10+5Β» 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 |