Development of ultra-sensitive Ca2+ indicators, yellow cameleon-nanos

Takeharu Nagai; Kazuki Horikawa; Tomoki Matsuda; Kentaro Kobayashi

doi:10.1117/12.871202

11 February 2011 Development of ultra-sensitive Ca²⁺ indicators, yellow cameleon-nanos

Takeharu Nagai, Kazuki Horikawa, Tomoki Matsuda, Kentaro Kobayashi

Proceedings Volume 7910, Reporters, Markers, Dyes, Nanoparticles, and Molecular Probes for Biomedical Applications III; 79100G (2011) https://doi.org/10.1117/12.871202
Event: SPIE BiOS, 2011, San Francisco, California, United States

Abstract

There are so many Ca²⁺ indicators such as Oregon green 488 BAPTA-1 and cameleon, all of which have moderate Ca²⁺ affinities (K_d > 150 nM). These indicators successfully report changing in Ca²⁺ concentration in most of cells. However, it is known that some cells are estimated to have a very low resting [Ca²⁺] and display very small ▵[Ca²⁺]s, which is below the detection limit of the indicators. To develop high-affinity indicators, we modified a FRET-based Ca²⁺ indicator, yellow cameleon (YC) 2.60 (K_d = 100 nM) because of the brightness and the large dynamic range. By engineering the Ca²⁺ sensing domain, we obtained a series of ultra-sensitive Ca²⁺ indicators. Their high Ca²⁺ affinities (Kd = 15, 30, and 50 nM) enabled detection of subtle Ca²⁺ transients associated with spontaneous network activity in zebrafish. Our measurements revealed that both the resting Ca²⁺ level and the amplitude of Ca²⁺ transients significantly differed by cell type and stimulation, indicating that the selection of indicators with the appropriate K_d is essential for successful in vivo Ca²⁺ imaging. A lineup of such indicators with finely tuned Kd values optimized to detect [Ca²⁺] from 10 nM to 100 nM would enable precise and reliable Ca²⁺ imaging.

Citation Download Citation

Takeharu Nagai, Kazuki Horikawa, Tomoki Matsuda, and Kentaro Kobayashi "Development of ultra-sensitive Ca²⁺ indicators, yellow cameleon-nanos", Proc. SPIE 7910, Reporters, Markers, Dyes, Nanoparticles, and Molecular Probes for Biomedical Applications III, 79100G (11 February 2011); https://doi.org/10.1117/12.871202

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

Members: $17.00

Non-members: $21.00 ADD TO CART

PROCEEDINGS
7 PAGES

DOWNLOAD PAPER SAVE TO MY LIBRARY

GET CITATION

RIGHTS & PERMISSIONS

Get copyright permission Get copyright permission on Copyright Marketplace

KEYWORDS

Calcium

Content addressable memory

Fluorescence resonance energy transfer

Proteins

In vivo imaging

Signal detection

Neurons

Show All Keywords

Keywords/Phrases

Search In:

Publication Years