Internal Perfusion

Port-a-Patch® – Reaching inside the cell.

•Robust and reliable internal solution exchange•Complete exchange of solutions in seconds•Continuous internal perfusion possible•Data recordings possible during solution exchange

1422

The Internal Perfusion System ispossibility to perfuse the intracellular side of thean add-on for the ­Port-a-Patch®.

membrane. This aspect is exploited with the Internal

Combined with Nanion’s Perfusion

Perfusion System for the Port-a-Patch®. Up to eight

Systemdifferent solutions can be perfused inside the cell,

either manually, or fully computer

allowing dose response analysis of compounds actingcontrolled.

on the cytosolic portion of the ion channel. Second

acquisition can be synchronized

messengers and metabolites can also be addedwith solution exchange, and tracesinternally to investigate how ion channel function isare automatically tagged with theaffected. Solutions are exchanged within seconds,compound information.

and the ion channel modulation can be continuously
monitored during the solution exchange.

15
23

it

canIn

be

thisoperated
way,

data

Internal Perfusion

Nanion’s planar patch clamp chips offer the unique400 pA% Inhibition

50 msK+10080

Stable recordings during internal ­perfusion.
KV1.3 currents (blue), ­endogenously ex-6040

pressed in Jurkat cells, were rapidly ­blocked

20

by internal perfusion of Cs+ (light blue), and01000
0.1
10Quinidine Concentration (µM)

Peak Amplitude (nA)

Cs+
1.5

K+

fully recovered after washout with K+ (grey).
Internal solution replacement was ­repeated
19 times and the recording was stable

1.0for over 35 minutes, as shown in the lower

0.5­graph.
Cs+05101520Time (min)

2530

35

Internal application of second messengers.
Currents mediated by BK channels expressed
2 nA

in CHO cells were studied using the Internal

500 msElevated Ca2+

Perfusion System. Currents were elicited by avoltage step from -80 mV to +80 mV before
andafteradding

aninternalsolution­containing a higher concentration of free

Current Amplitude (pA)

0 µM Ca2+

Capsaicin (External)

800

Ca2+.

Internal

applicationofCa2+-calmodulin

to TRPV1.
TRPV1 currents were elicited by the ­external

600

application of 20 μM capsaicin. After

400
200

CaM (Internal)­capsaicin activation, the currents wereControl­partially blocked by the internal applicationof Ca2+-calmodulin (50 μM Ca2+ / 500 nMCaM). TRPV1 channels were expressed in0CHO cells.

-2000.00

0.05

0.10

0.15 0.20
Time (s)

0.25

0.30

0.35

Rapid internal exchange.
5 pA

1s

Switchingofinternal­solutions

during­gramicidin recordings from a ­lipid ­bilayer
was obtained within seconds. A lipid bilayer
was formed using ­giant ­unilamellar vesicles.
­Gramicidin was incorporated into the lipidbilayer. ­Currents were recorded at a holding­potential of +150 mV. The internal solutionwas switched from HCl to KCl, ­resulting in­lower channel conductance.

100 mM HCl

100 mM KCl

1424Pharmacological experiments.

of quinidine (left) or TEA+ (right). The graphs
show the Hill plots for IC50 determination. For
quinidine the IC50 was determined as 15 µM(literature value external application 14 µM)
and for TEA the IC50 was determined as 0.9+± 0.3 mM (n = 3) (literature value internal

Normalised Blockapplication of increasing concentrations

1001.0800.8

Normalised Block

KV1.3 current was blocked by the internal

60402000.40.2
0.00.1­application 0.6 mM).

11010010000.010.1+Product Number

Specification

Size & Weight

02 2001

Internal Perfusion with Perfusion System (02 1002)

Valve Control Panel
Size (l x w x h):
40 x 5.8 x 13 cm
Weight: 2.45 kg

02 2002

Internal Perfusion without Perfusion System*• Internal Perfusion Chamber with housing• Internal Perfusion starter kit: electrodes, Reagent Kit,
tubings and other accessories* Only compatible with Nanion’s Perfusion System

1TEA (mM)

Quinidine (µM)•V alve Control Panel (analog, digital and USB-ports forcomputer control)• Internal Perfusion Chamber with housing• Internal Perfusion starter kit: electrodes, solutions, tubingsand other accessories•P erfusion System starter kit with 50 ml syringes, Reagent Kit,
tubing, connectors, waste bottle etc.
• S tand with syringe holder

15
25

0.6

Internal perfusion
Size (l x w x h):
7.0 x 7.0 x 10 cm
Weight: 460 g
Internal perfusion
Size (l x w x h):
7.0 x 7.0 x 10 cm
Weight: 460 g

10100