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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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References

Agrawal, A, Gupta, M, Veeraraghavan, A & Narasimhan, SG (2010). Optimal coded sampling for temporal super-resolution. In Proceedings of the IEEE Computer Society Conference on Computer Vision and Pattern Recognition, pp. 599–626. San Francisco, CA: IEEE.Google Scholar
Bennett, MR (1983). Development of neuromuscular synapses. Physiol Rev 63(3), 915–1048.CrossRefGoogle ScholarPubMed
Bullen, A, Patel, SS & Saggau, P (1997). High-speed, random-access fluorescence microscopy: I. High-resolution optical recording with voltage-sensitive dyes and ion indicators. Biophys J 73(1), 477–491.CrossRefGoogle ScholarPubMed
Bullen, A & Saggau, P (1998). Indicators and optical configuration for simultaneous high-resolution recording of membrane potential and intracellular calcium using laser scanning microscopy. Pflug Arch Eur J Phys 436(5), 788–796.CrossRefGoogle ScholarPubMed
Bullen, A & Saggau, P (1999 a). High-speed, random-access fluorescence microscopy: II. Fast quantitative measurements with voltage-sensitive dyes. Biophys J 76(4), 2272–2287.CrossRefGoogle ScholarPubMed
Bullen, A & Saggau, P (1999 b). Optical recording from individual neurons in culture. In Modern Techniques in Neuroscience Research, Windhorst, U & Johansson, H (Eds.), pp. 89–126. Berlin, Heidelberg: Springer.CrossRefGoogle Scholar
Chan, KG, Streichan, SJ, Trinh, LA & Liebling, M (2016). Simultaneous temporal superresolution and denoising for cardiac fluorescence microscopy. IEEE Trans Comput Imag 2(3), 348–358.CrossRefGoogle Scholar
Cohen, LB & Salzberg, BM (1978). Optical measurement of membrane potential. In Reviews of Physiology, Biochemistry and Pharmacology, vol. 83, pp. 35–88. Berlin, Heidelberg: Springer.Google Scholar
Cox, G (2002). Biological confocal microscopy. Mater Today 5(3), 34–41.CrossRefGoogle Scholar
Grinnell, AD & Trussell, LO (1983). Synaptic strength as a function of motor unit size in the normal frog sartorius. J Physiol 338(1), 221–241.CrossRefGoogle ScholarPubMed
Kazakov, A, Alexandrov, M, Zhilyakov, NV, Khaziev, EF & Samigullin, DV (2015). A simple suction electrode for electrical stimulation of biological objects. Int Res J 9(40), 13–16.Google Scholar
Khaziev, E, Golovyahina, A, Bukharaeva, E, Nikolsky, E & Samigullin, D (2017). Action of ATP on Ca2+-transient in different parts of the frog motor nerve ending. BioNanoScience 7(1), 254–257.CrossRefGoogle Scholar
Land, BR, Johnson, BR, Wyttenbach, RA & Hoy, RR (2004). Tools for physiology labs: Inexpensive equipment for physiological stimulation. J Undergrad Neurosci Educ 3, 1.Google ScholarPubMed
Llinas, R, Steinberg, IZ & Walton, K (1976). Presynaptic calcium currents and their relation to synaptic transmission: Voltage clamp study in squid giant synapse and theoretical model for the calcium gate. Proc Natl Acad Sci USA 73(8), 2918–2922.CrossRefGoogle ScholarPubMed
Mallart, A (1984). Presynaptic currents in frog motor endings. Pflugers Arch 400(1), 8–13.CrossRefGoogle ScholarPubMed
Mukhitov, A, Arkhipova, S & Nikolsky, E (2011). Modern Light Microscopy in Biological and Medical Research, 1st edn. Moscow: Nauka.Google Scholar
Nikol'skii, EE, Bukharaeva, ÉA, Samigullin, DV & Gainulov, RK (2002). Characteristics of the time course of evoked secretion of transmitter quanta in different parts of the motor nerve ending in the frog. Neurosci Behav Physiol 32(3). doi:10.1023/A:1015010307181CrossRefGoogle Scholar
Rama, S (2015). Shift and mean algorithm for functional imaging with high spatio-temporal resolution. Front Cell Neurosci 9. doi:10.3389/fncel.2015.00446CrossRefGoogle ScholarPubMed
Ryabinin, YA (1968). Stroboscopic Oscillography of Nanosecond Signals, 2nd edn. Moscow: Soviet Radio.Google Scholar
Samigullin, D, Fatikhov, N, Khaziev, E, Skorinkin, A, Nikolsky, E & Bukharaeva, E (2015). Estimation of presynaptic calcium currents and endogenous calcium buffers at the frog neuromuscular junction with two different calcium fluorescent dyes. Front Synaptic Neurosci 6. doi:10.3389/fnsyn.2014.00029CrossRefGoogle ScholarPubMed
Samigullin, DV, Arkhipov, AY & Khaziev, EF (2018). Fast fluorescent signals with the confocal laser scanning microscope recording method, Russia. Retrieved from http://www1.fips.ru/fips_servl/fips_servlet?DB=RUPAT&DocNumber=2680664&TypeFile=htmlGoogle Scholar
Samigullin, DV, Khaziev, EF, Zhilyakov, NV, Bukharaeva, EA & Nikolsky, EE (2017). Loading a calcium dye into frog nerve endings through the nerve stump: Calcium transient registration in the frog neuromuscular junction. J Vis Exp 2017(125). doi:10.3791/55122Google Scholar
Samigullin, DV, Vasin, AL, Bukharaeva, EA & Nikolsky, EE (2010). Characteristics of calcium transient in different parts of frog nerve terminal in response to nerve impulse. Dokl Biol Sci 431(1). doi:10.1134/S0012496610020043CrossRefGoogle ScholarPubMed
Shechtman, E, Caspi, Y & Irani, M (2005). Space-time super-resolution. IEEE Trans Pattern Anal Mach Intell 27(4), 531–545.CrossRefGoogle ScholarPubMed
Toomre, D & Pawley, JB (2006). Disk-scanning confocal microscopy. In Handbook of Biological Confocal Microscopy, Pawley, JB (Ed.), 3rd ed. pp. 221–238.CrossRefGoogle Scholar
Tsien, RY (1988). Fluorescence measurement and photochemical manipulation of cytosolic free calcium. Trends Neurosci 11(10), 419–424.CrossRefGoogle ScholarPubMed
Tsien, RY (1989). Fluorescent indicators of ion concentrations. Methods Cell Biol 30(C), 127–156.CrossRefGoogle ScholarPubMed
Tsien, RY & Waggoner, A (1995). Fluorophores for confocal microscopy. In Handbook of Biological Confocal Microscopy, Pawley, JB (Eds.), pp. 267–279. Boston, MA: Springer.CrossRefGoogle Scholar
Veeraraghavan, A, Reddy, D & Raskar, R (2011). Coded strobing photography: Compressive sensing of high speed periodic videos. IEEE Trans Pattern Anal Mach Intell 33(4), 671–686.CrossRefGoogle ScholarPubMed
Webb, RH (1996). Confocal optical microscopy. Rep Prog Phys 59(3), 427–471.CrossRefGoogle Scholar
Wilson, T (1990. Optical aspects of confocal microscopy. In Confocal Microscopy, Wilson, T (Ed.), pp. 93–141. Oxford: Pergamon Press.Google Scholar
Yuste, R & Konnerth, A (2005). Imaging in neuroscience and development, a laboratory manual. Cold Spring Harbor, NY: Cold Spring Harbor Laboratory Press.Google Scholar
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