Dynamic ratiometric imaging of cytosolic free Ca2+ in skeletal muscle cells using 340/385-nm light-emitting diode illuminators
(2018) In IEEE Photonics Journal 10(6).- Abstract
Calcium-sensitive fluorescent indicators fall broadly into two categories, ratiometric (dual-wavelength) or single-wavelength indicators based on their response to a calcium elevation. Ratiometric indicators shift either their excitation or their emission wavelengths in response to calcium, allowing the concentration of intracellular calcium to be determined from the ratio of fluorescence emission or excitation at distinct wavelengths. Fura-2 is one of the most common ratiometric fluorescent Ca2+ indicators, which has an emission maximum at 510 nm whereas its excitation maximum changes from 380 to 340 nm in response to calcium binding. Historically, a combination of arc lamp and monochromators has been used as the light... (More)
Calcium-sensitive fluorescent indicators fall broadly into two categories, ratiometric (dual-wavelength) or single-wavelength indicators based on their response to a calcium elevation. Ratiometric indicators shift either their excitation or their emission wavelengths in response to calcium, allowing the concentration of intracellular calcium to be determined from the ratio of fluorescence emission or excitation at distinct wavelengths. Fura-2 is one of the most common ratiometric fluorescent Ca2+ indicators, which has an emission maximum at 510 nm whereas its excitation maximum changes from 380 to 340 nm in response to calcium binding. Historically, a combination of arc lamp and monochromators has been used as the light source for Fura-2 ratiomatric fluorescence microscopy. In recent years, different combinations of LEDs such as 350/380 nm or 360/380 nm have been used to excite Fura-2. To precisely match the Fura-2 excitation, in this study, we built a new fast switchable 340/385-nm LED excitation light source (Prizmatix Ltd., Israel). The newly constructed light source has been exploited in Fura-2 ratiometric calcium imaging of skeletal muscle cells. The spontaneously elicited Ca2+ transients in the cells were recorded with high temporal resolution. The light source utilized for the demonstrated instrumentation in this report optimally matches the excitation wavelengths of either calcium-free or bound states of Fura-2. The high-intensity stability and fast switching of the 340/385-nm LED illuminators indicate their potential as a preferred light source for Fura-2 ratiometric calcium imaging over the existing light sources.
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- author
- Chenchiliyan, Manoop LU ; Portal, Dana Adler ; Chakraborty, Ruchira ; Ben-Gal, Tal Shahar ; Deutsch, Assaf ; Pewzner, Eliahu ; Shainberg, Asher ; Duadi, Hamootal and Fixler, Dror
- publishing date
- 2018-12
- type
- Contribution to journal
- publication status
- published
- subject
- keywords
- calcium imaging, fluorescence, Fura-2, LED, microscopy, rat myotubes, Ratio-metric imaging
- in
- IEEE Photonics Journal
- volume
- 10
- issue
- 6
- article number
- 8540908
- publisher
- IEEE - Institute of Electrical and Electronics Engineers Inc.
- external identifiers
-
- scopus:85057206216
- ISSN
- 1943-0655
- DOI
- 10.1109/JPHOT.2018.2882503
- language
- English
- LU publication?
- no
- additional info
- Publisher Copyright: © 2018 IEEE.
- id
- c445ca3f-bb56-442f-8eca-5648d873580f
- date added to LUP
- 2023-06-16 10:40:32
- date last changed
- 2023-06-29 10:30:09
@article{c445ca3f-bb56-442f-8eca-5648d873580f,
abstract = {{<p>Calcium-sensitive fluorescent indicators fall broadly into two categories, ratiometric (dual-wavelength) or single-wavelength indicators based on their response to a calcium elevation. Ratiometric indicators shift either their excitation or their emission wavelengths in response to calcium, allowing the concentration of intracellular calcium to be determined from the ratio of fluorescence emission or excitation at distinct wavelengths. Fura-2 is one of the most common ratiometric fluorescent Ca<sup>2+</sup> indicators, which has an emission maximum at 510 nm whereas its excitation maximum changes from 380 to 340 nm in response to calcium binding. Historically, a combination of arc lamp and monochromators has been used as the light source for Fura-2 ratiomatric fluorescence microscopy. In recent years, different combinations of LEDs such as 350/380 nm or 360/380 nm have been used to excite Fura-2. To precisely match the Fura-2 excitation, in this study, we built a new fast switchable 340/385-nm LED excitation light source (Prizmatix Ltd., Israel). The newly constructed light source has been exploited in Fura-2 ratiometric calcium imaging of skeletal muscle cells. The spontaneously elicited Ca<sup>2+</sup> transients in the cells were recorded with high temporal resolution. The light source utilized for the demonstrated instrumentation in this report optimally matches the excitation wavelengths of either calcium-free or bound states of Fura-2. The high-intensity stability and fast switching of the 340/385-nm LED illuminators indicate their potential as a preferred light source for Fura-2 ratiometric calcium imaging over the existing light sources.</p>}},
author = {{Chenchiliyan, Manoop and Portal, Dana Adler and Chakraborty, Ruchira and Ben-Gal, Tal Shahar and Deutsch, Assaf and Pewzner, Eliahu and Shainberg, Asher and Duadi, Hamootal and Fixler, Dror}},
issn = {{1943-0655}},
keywords = {{calcium imaging; fluorescence; Fura-2; LED; microscopy; rat myotubes; Ratio-metric imaging}},
language = {{eng}},
number = {{6}},
publisher = {{IEEE - Institute of Electrical and Electronics Engineers Inc.}},
series = {{IEEE Photonics Journal}},
title = {{Dynamic ratiometric imaging of cytosolic free Ca<sup>2+</sup> in skeletal muscle cells using 340/385-nm light-emitting diode illuminators}},
url = {{http://dx.doi.org/10.1109/JPHOT.2018.2882503}},
doi = {{10.1109/JPHOT.2018.2882503}},
volume = {{10}},
year = {{2018}},
}