Literature DB >> 25561614

Verifying the function and localization of genetically encoded Ca2+ sensors and converting FRET ratios to Ca2+ concentrations.

J Genevieve Park1, Amy E Palmer1.   

Abstract

Genetically encoded, ratiometric, fluorescent Ca(2+) biosensors can be used in living cells to quantitatively measure free Ca(2+) concentrations in the cytosol or in organelles. This protocol describes how to perform a calibration of a Ca(2+) sensor expressed in cultured mammalian cells as images are acquired using a widefield fluorescence microscope. This protocol also explains how to calculate Förster resonance energy transfer (FRET) ratios from acquired images and how to convert FRET ratios to Ca(2+) concentrations.
© 2015 Cold Spring Harbor Laboratory Press.

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Year:  2015        PMID： 25561614      PMCID： PMC4968932          DOI： 10.1101/pdb.prot076547

Source DB:  PubMed          Journal:  Cold Spring Harb Protoc        ISSN： 1559-6095


  12 in total

1.  Expanded dynamic range of fluorescent indicators for Ca(2+) by circularly permuted yellow fluorescent proteins.

Authors:  Takeharu Nagai; Shuichi Yamada; Takashi Tominaga; Michinori Ichikawa; Atsushi Miyawaki
Journal:  Proc Natl Acad Sci U S A       Date:  2004-07-09       Impact factor: 11.205

2.  Spontaneous network activity visualized by ultrasensitive Ca(2+) indicators, yellow Cameleon-Nano.

Authors:  Kazuki Horikawa; Yoshiyuki Yamada; Tomoki Matsuda; Kentarou Kobayashi; Mitsuhiro Hashimoto; Toru Matsu-ura; Atsushi Miyawaki; Takayuki Michikawa; Katsuhiko Mikoshiba; Takeharu Nagai
Journal:  Nat Methods       Date:  2010-08-08       Impact factor: 28.547

3.  Ca2+ indicators based on computationally redesigned calmodulin-peptide pairs.

Authors:  Amy E Palmer; Marta Giacomello; Tanja Kortemme; S Andrew Hires; Varda Lev-Ram; David Baker; Roger Y Tsien
Journal:  Chem Biol       Date:  2006-05

4.  Circularly permuted green fluorescent proteins engineered to sense Ca2+.

Authors:  T Nagai; A Sawano; E S Park; A Miyawaki
Journal:  Proc Natl Acad Sci U S A       Date:  2001-03-06       Impact factor: 11.205

5.  A new generation of Ca2+ indicators with greatly improved fluorescence properties.

Authors:  G Grynkiewicz; M Poenie; R Y Tsien
Journal:  J Biol Chem       Date:  1985-03-25       Impact factor: 5.157

6.  Bcl-2-mediated alterations in endoplasmic reticulum Ca2+ analyzed with an improved genetically encoded fluorescent sensor.

Authors:  Amy E Palmer; Can Jin; John C Reed; Roger Y Tsien
Journal:  Proc Natl Acad Sci U S A       Date:  2004-12-07       Impact factor: 11.205

7.  A genetically encoded calcium indicator for chronic in vivo two-photon imaging.

Authors:  Marco Mank; Alexandre Ferrão Santos; Stephan Direnberger; Thomas D Mrsic-Flogel; Sonja B Hofer; Valentin Stein; Thomas Hendel; Dierk F Reiff; Christiaan Levelt; Alexander Borst; Tobias Bonhoeffer; Mark Hübener; Oliver Griesbeck
Journal:  Nat Methods       Date:  2008-09       Impact factor: 28.547

8.  Mechanisms of control of the free Ca2+ concentration in the endoplasmic reticulum of mouse pancreatic β-cells: interplay with cell metabolism and [Ca2+]c and role of SERCA2b and SERCA3.

Authors:  Magalie A Ravier; Dorothée Daro; Leticia Prates Roma; Jean-Christophe Jonas; Rui Cheng-Xue; Frans C Schuit; Patrick Gilon
Journal:  Diabetes       Date:  2011-09-01       Impact factor: 9.461

9.  Who needs microtubules? Myogenic reorganization of MTOC, Golgi complex and ER exit sites persists despite lack of normal microtubule tracks.

Authors:  Kristien J M Zaal; Ericka Reid; Kambiz Mousavi; Tan Zhang; Amisha Mehta; Elisabeth Bugnard; Vittorio Sartorelli; Evelyn Ralston
Journal:  PLoS One       Date:  2011-12-27       Impact factor: 3.240

10.  Measuring steady-state and dynamic endoplasmic reticulum and Golgi Zn2+ with genetically encoded sensors.

Authors:  Yan Qin; Philip J Dittmer; J Genevieve Park; Katarina B Jansen; Amy E Palmer
Journal:  Proc Natl Acad Sci U S A       Date:  2011-04-18       Impact factor: 11.205
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  1 in total

Review 1.  Nanoparticle-Based and Bioengineered Probes and Sensors to Detect Physiological and Pathological Biomarkers in Neural Cells.

Authors:  Dusica Maysinger; Jeff Ji; Eliza Hutter; Elis Cooper
Journal:  Front Neurosci       Date:  2015-12-18       Impact factor: 4.677
  1 in total

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