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Enhancement of the Temporal Resolution of Fluorescent Signals Acquired by the Confocal Microscope

Published online by Cambridge University Press:  02 March 2020

Arsenii Y. Arkhipov ,
Eduard F. Khaziev ,
Andrey I. Skorinkin ,
Ellya A. Bukharaeva  and
Dmitry V. Samigullin Open the ORCID record for Dmitry V. Samigullin [Opens in a new window]
Arsenii Y. Arkhipov
Affiliation:
Laboratory of Biophysics of Synaptic Processes, Kazan Institute of Biochemistry and Biophysics, FRC Kazan Scientific Center of RAS, 420111Kazan, Russia Department of Radiophotonics and Microwave Technologies, Kazan National Research Technical University named after A.N. Tupolev, 420111Kazan, Russia
Eduard F. Khaziev
Affiliation:
Laboratory of Biophysics of Synaptic Processes, Kazan Institute of Biochemistry and Biophysics, FRC Kazan Scientific Center of RAS, 420111Kazan, Russia Department of Radiophotonics and Microwave Technologies, Kazan National Research Technical University named after A.N. Tupolev, 420111Kazan, Russia Open Laboratory of Neuropharmacology, Kazan Federal University, 420111Kazan, Russia
Andrey I. Skorinkin
Affiliation:
Laboratory of Biophysics of Synaptic Processes, Kazan Institute of Biochemistry and Biophysics, FRC Kazan Scientific Center of RAS, 420111Kazan, Russia
Ellya A. Bukharaeva
Affiliation:
Laboratory of Biophysics of Synaptic Processes, Kazan Institute of Biochemistry and Biophysics, FRC Kazan Scientific Center of RAS, 420111Kazan, Russia
Dmitry V. Samigullin*
Affiliation:
Laboratory of Biophysics of Synaptic Processes, Kazan Institute of Biochemistry and Biophysics, FRC Kazan Scientific Center of RAS, 420111Kazan, Russia Department of Radiophotonics and Microwave Technologies, Kazan National Research Technical University named after A.N. Tupolev, 420111Kazan, Russia
*
*Author for correspondence: Dmitry Samigullin, E-mail: [email protected]
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Abstract

Here, we describe a method of acquisition of fast fluorescent signals with the help of the laser scanning confocal microscope (LSCM). Our method permits an increase in the temporal resolution of acquired signals. The method is based on LSCM recordings of fast fluorescent signals with the shortest achievable time sweep, which are performed with the help of a proprietary algorithm. A series of recordings is made in multiple steps; at each step, the fluorescent signal is incremented by a time interval smaller than the time sweep of the frame of LSCM. The size of the increment determines the achievable time resolution. The convolution of the recorded images results in a signal with the temporal resolution determined by the chosen time increment. This method was applied to register the change in fluorescence (calcium transient) of calcium dye preloaded into peripheral nerve endings by electrical stimulation of the motor nerve. Calculated parameters of the calcium transient were identical to the parameters obtained earlier with the help of a high-speed camera and photodiode. We conclude that the method described here can be applied for the registration of fast fluorescent signals by LSCM with a high spatial and temporal resolution.

Keywords

calcium transientconfocal microscopyimage acquisitionmolecular and cellular imagingtemporal resolution
Type
Software and Instrumentation
Information
Microscopy and Microanalysis , Volume 26 , Issue 2 , April 2020 , pp. 204 - 210
Copyright
Copyright © Microscopy Society of America 2020

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