Imaging of calcium transients in cultured neurons by TCSPC FLIM (Conference Presentation)

Wolfgang Becker; Samuel Frere

doi:10.1117/12.2214083

28 April 2016 Imaging of Ca²⁺ transients in cultured neurons by temporal mosaic FLIM (Conference Presentation)

Wolfgang Becker, Samuel Frere

Proceedings Volume 9712, Multiphoton Microscopy in the Biomedical Sciences XVI; 97120O (2016) https://doi.org/10.1117/12.2214083
Event: SPIE BiOS, 2016, San Francisco, California, United States

Abstract

We present a technique that records transient changes in the concentration of free calcium in live neurons by TCSPC FLIM. The sample is incubated with a calcium-sensitive dye. To measure the temporal change in the calcium-ion concentration the sample is periodically stimulated by an electrical signal and scanned at high image rate with a high-frequency pulsed laser beam. Single photons of the fluorescence light are detected, and a photon distribution over the coordinates of the scan, the arrival times of the photons after the excitation pulses, and the time after the stimulation pulses is built up. The result can be interpreted as a sequence of FLIM images for different times after the stimulation pulses. The signal-to-noise ratio only depends on the available photon rate and the total acquisition time, not on the speed of the sequence. The maximum resolution at which lifetime changes can be recorded is given by the frame rate of the scanner which is currently 38 ms. Faster changes can be recorded by line scanning. Transient lifetime effects can then be resolved at a resolution of about one millisecond.

Conference Presentation

Citation Download Citation

Wolfgang Becker and Samuel Frere "Imaging of Ca²⁺ transients in cultured neurons by temporal mosaic FLIM (Conference Presentation)", Proc. SPIE 9712, Multiphoton Microscopy in the Biomedical Sciences XVI, 97120O (28 April 2016); https://doi.org/10.1117/12.2214083

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
PRESENTATION

WATCH
PRESENTATION SAVE TO MY LIBRARY

GET CITATION

KEYWORDS

Fluorescence lifetime imaging

Calcium

Neurons

Fluorescence spectroscopy

Luminescence

Photodetectors

Scanners

Show All Keywords

Keywords/Phrases

Search In:

Publication Years