אלקטרו הקלטה בו זמנית ובסידן הדמיה של suprachiasmatic גרעין נוירונים

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Irwin, R. P., Allen, C. N. Simultaneous Electrophysiological Recording and Calcium Imaging of Suprachiasmatic Nucleus Neurons. J. Vis. Exp. (82), e50794, doi:10.3791/50794 (2013).

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Abstract

Materials

Name Company Catalog Number Comments
NaCl Fisher Scientific Co. S271-3
KCl Fisher Scientific Co. P217-500
NaH2PO4•H2O Sigma Chemical Co. S-9638
MgCl2•6H2O Fisher Scientific Co. M33-500
CaCl2•2H2O Fisher Scientific Co. C79-500
D-glucose Fisher Scientific Co. D16-500
NaHCO3 Fisher Scientific Co. S233-500
Sucrose Fisher Scientific Co. S5-500
Potassium D-gluconate Sigma-Aldrich G4500
HEPES Sigma-Aldrich H4034
Adenosine 5′-triphosphate dipotassium salt dihydrate Sigma-Aldrich A8937
Guanosine 5′-triphosphate tris salt Sigma-Aldrich G9002
Agarose (ultrapure) Life Technologies 15510-027 Gel pored into sterile Petri dish 6 mm thick layer
KOH Sigma-Aldrich P-6310
NaOH Sigma-Aldrich S-5881
Bis-fura-2, hexapotassium salt Invitrogen B6810 Cell impermeant
Name of Equipment Company Catalog Number Comments
Microtome Leica VT1000S Tissue slicing
Cyanoacrylic glue (Roti-Coll1) Carl Roth GmbH+Co Art-Nr. 0258.1
Microelectrode puller Narshige International USA PP-83
Microelectrode Capillary Tubes World Precision Instruments 1B150F-4
Microfil 34 g World Precision Instruments MF34G-5
Syringe filter Corning #431212
Microscope Leica DM LFS With 4X, 40X UV and 63X UV objectives, and epifluorescence
CCD Camera Hamamatsu ORCA ER 12 bit CCD
Fura-2 Filter Cube Chroma 71500A Set with UG11 filter
Polychrome IV Till Photonics GmBH Monochronometer
Ultraviolet blocking safety glasses Ultra-Violet Products
EPC-9 amplifier HEKA Eletronik
Metafluor Molecular Devices Imaging Software
Patchmaster HEKA Eletronik Data Acquisition Software
Igor version 6 Wavemetrics Electrophysiology & Ca2+ Data Analysis
VAPRO 5520 Westcor Vapor pressure osmometer

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References

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