Methodenartikel

Establishing a Whole-Cell Patch Clamp for Electrophysiological Recording from a Flat-Mount Mouse Retina

8 juli 2025

In dit artikel

Samenvatting

$$\rightleftharpoonup{xx}$$ $$\longleftharp{xx}$$, $$\longrightharp{xx}$$,

Source: Tu, H., et al. Patch Clamp Recording of Starburst Amacrine Cells in a Flat-mount Preparation of Deafferentated Mouse Retina. J. Vis. Exp. (2016).

This video demonstrates a whole-cell patch-clamp technique for electrophysiological recording from a flat-mount mouse retina. A thin glass pipette with an electrode is used to approach starburst amacrine cells (SACs) within the retina. After forming a seal, suction is applied to rupture the membrane and form the whole-cell patch-clamp configuration. Finally, the flow of ions through the SAC synaptic receptors is measured using the configuration.

Protocol

$$\rightleftharpoonup{xx}$$ $$\longleftharp{xx}$$, $$\longrightharp{xx}$$,

All procedures involving sample collection have been performed in accordance with the institute's IRB guidelines.

1. External and Internal Solutions

  1. Use mammalian Ringer's solution during retina dissection and as the external solution in subsequent electrophysiological recording. Prepare the mammalian Ringer's solution from 10x stock solution (without calcium) on the day of recording, and add CaCl2 drop-wise after 15 min of carbogenation (95% O2 and 5% CO2). The final 1x solution contains (in mM): 120 NaCl, 5 KCl, 25 NaHCO3, 0.8 Na2HPO4, 0.1 NaH

Toegang beperkt. Log in of start een proefperiode om deze inhoud te bekijken.

Openbaarmakingen

$$\rightleftharpoonup{xx}$$ $$\longleftharp{xx}$$, $$\longrightharp{xx}$$,
No conflicts of interest declared.

Materialen

Lijst van materialen gebruikt in dit artikel
NaamBedrijfCatalogusnummerOpmerkingen
Fixed-stage fluorescent microscope with DICOlympusBX51-WI
MicromanipulatorsSutterMP-225
Patch clamp amplifierA-M SystemAM2400
AD converterNational InstrumentNI-USB-6221
Heater controllerWarner InstrumentTC-324B
Inline heaterWarner InstrumentSC-20
Peristaltic pumpRaininDynamax
pipette pullerSutter InstrumentP-1000
Glass tube with filamentKing Precision GlassCustomized
StimulatorA.M.P.I.Master-8
BiocytinSigmaB4261
NaClSigmaS6191
KClSigmaP5405
NaHCO3FisherBP328-1
Na2HPO4SigmaS0876
NaH2PO4SigmaS5011
CaCl2SigmaC5670
MgSO4SigmaM1880
D-glucoseSigmaG6152
K-gluconateSigmaG4500
ATP-MgSigmaA9187
Li-GTPSigmaG5884
EGTASigmaE0396
HEPESSigmaH4034
KOHSigmaP5958
Cs-methanesulfonateSigmaC1426
CsOHSigma232041
Syringe filterNalgene171
1 ml syringeRainin17013002
10 ul pipette tipGenesee Scientific24-130RL
10X PBSLonza17-517Q

Herprints en machtigingen

Toestemming aanvragen om de tekst of afbeeldingen van dit JoVE-artikel te hergebruiken

Toestemming aanvragen

Trefwoorden

Starburst amacrinecellenflat mount preparatiepipetweerstandGiga ohm zegelspanningsclampmodusmeting van synaptische stroomelektrodehouder

Gerelateerde artikelen