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Ussing chamber for Costar filters.
Filter types.
Three different chambers are available.
Costar filter types: - Transwell 12 mm
- Transwell 6.5 mm
- Snapwell 12 mm
- surface area: 1.13 cm2.
- surface area: 0.33 cm2.
- surface area: 1.13 cm2.
Except for the main part (MP-65, MP12 and MP-SW), all parts of the chamber
assembly are identical and interchangeable.
Operation modes.
Each compartment can be perfused continuously or experiments can be performed in
stopped flow mode.
Continuous perfusion: Solutions can be directly pumped out of both chamber
chamber compartments. Alternatively, outflow can be driven by gravity through the
solution outflow port, thus avoiding vibrations and associated noise. Recirculation of
the apical as well as basolateral perfusate can be applied by using small peristaltic
pumps.
Stopped flow mode: The chamber design enables gas bubbling of the solution in the
apical as well as basolateral compartment.
Figure 1: Open assembly of the Ussing chamber. Covers (apical and basolateral) are simply inserted in the
main part and attached by a bayonet fitting. Sealing between AP and BL side of the main part: O-ring around the
shaft of the Costar filter. All ports have the diameter so that the different connections can be inserted.
Filter insertion.
The Costar transwell filters are inserted in the apical chamber upper halve of the
main part and held down by the cover that presses on the upper rim of the filter.
Sealing between AP and BL side is performed with an O-ring that resides in the
chamber and fits around the shaft of the filter.
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Characteristics.
As depicted in Fig. 1, the chamber consists of 3 parts in which electrodes and
perfusion tubes can be inserted. All inserted parts have the same diameter (6 mm)
as well as all ports. Unused ports can be closed by dummy Lucite closures foreseen
of the same O-ring as the electrodes and perfusion connections.
- With exception for the support (round plate and post), all parts are made of Lucite.
- All surfaces of the chamber parts are accessible for cleaning.
The basolateral cover is sealed with an O-ring.
- The chamber is mounted on a support that allows inclination of the chamber. The
inclination guarantees that air bubbles that enter the basolateral compartment
through the inflow tubing will not reach the filter membrane and escape through the
outflow port. The angle of inclination can be chosen by the user. An angle of about
6° should be sufficient.
The main part (light green in figures) is the only part that is specific for the type of
Costar filter: 12 or 6.5 mm filter. All other parts are interchangeable.
A
Figure 2: Ussing chamber with support (Costar transwell 6.5 mm version).
A: Side view of assembly with support.
B: Chamber with electrodes and perfusion in- and outlets.
Dimensions
Diameter of chamber: 54 mm
Total height with covers: 74 mm
Diameter of ports: 6 mm.
B
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Volume of chamber compartments
(ml)
Filter type
BL compartment
AP compartment (Top suction outflow)
AP in continuous (gravity) outflow mode
12 mm
3.2
1.2
2.0
6.5 mm
1.6
0.4
1.1
Inflow and outflow ports.
All ports (inflow, outflow and electrodes) have the same diameter (as well as the
electrodes). The user has the choice between 2 types of inflow parts. Both types
have peek tube connections on which standard Tygon tubing fits (ID 1.3 mm ID). An
O-ring provides sealing of the inflow as well as outflow connections.
Inflow type 1:
Peek tubing fits in Lucite holder with ferrule and can be changed by user.
Inside diameter (ID) of tubing: 0.75, 0.5 or 0.25 mm.
Outside diameter (OD) of tubing: 1.57 mm.
Inflow type 2:
This part is factory assembled and peek tubing can not be changed by the user.
Standard ID of peek tubing is 0.5 mm. 0.25 and 0.75 mm types are available on
request.
Outflow:
Stainless steel tubing glued in Lucite holder of 6 mm diameter
Electrodes
Current electrodes:
The assembly of a platinum wire connected to a gold plated contact pin is inserted in
a Lucite holder with a ferrule fitting. Sealing of the electrode in the chamber is
provided by an O-Ring. Instead of these Pt electrodes, the user can decide to use
the same Ag/AgCl pellet electrodes as used for voltage recording.
A
B
Figure 3: Electrodes.
A: current electrode with Pt wire.
B: voltage electrode with Ag/AgCl pellet and Agar bridges or closed KCl compartment..
Voltage electrodes:
Voltage electrodes are constructed with a Ag/AgCl pellet that is soldered to a gold
plated contact pin. Contact pin and AgCl pellet are factory assembled with peek
tubing and contact cover (green). This assembly is inserted in the Lucite holder that
contains KCl solution. The connection between the KCl solution and the perfusion
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solutions is made with the peek tubing. The inside diameter of the peek tubing
determines the resistance of the electrode and can be 0.125, 0.25, 0.5, 0.75 or 1
mm. Alternatively, the connection between the part with the KCl solution and the
bath can be made with polyethylene (PE) tubing filled with agar (agar bridge). To
main the shape, the PE tubing is supported by a thin walled polyimide tubing.
The Lucite holder is perfectly closed with the ferrule that fits around the Ag_AgCl
holding peek tubing. Assembling is very easy: the Lucite holder is filled with KCl, the
pellet is inserted and the assembly is closed by pressing the ferrule into the fitting of
the Lucite holder with the Lucite closure.
Part list
Reference code
Illustration
UC-65-CP
See Fig 2A
Ussing chamber, 6.5 mm, complete with support
UC-12-CP
Ussing chamber, 12 mm, complete with support
CS
Support
MP-65
Main part 6.5 mm
MP-12
Main part 12 mm
COV-AP
Cover AP compartment
See Fig 2A
5
COV-BL
Cover BL compartment
CE-AS
Current electrode complete assembly
CE-PT
Current electrode – Pt wire and plug – 1mm
CE-HD
Current electrode – Lucite holder
VE-AS
Voltage electrode - complete assembly
VE-AG
Voltage electrode – Ag/AgCl pellet with plug (1mm).
VE-HD
Voltage electrode – Holder and KCl compartment.
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IF-ADJ
Inflow adjustable length.
IF-FIX
Inflow – fixed length
OF
Ouflow connection
CL
Closure for unused 6 mm ports.
VC-CON
4 wires with colour coded sockets with wires for
connection to electronic device. Length: 30 cm
Not shown
Note: scaling of different parts is variable. For reference: all inserts with an O-ring
(red) have an outside diameter of 6 mm.
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Accessories
1 – Wire with 2 sockets (1mm) for interconnection of voltage or current electrodes.
Reference number: AS-WIRE
Price: 7.50 €
2 -.Container with cover for storing electrodes.
Dimensions: H: 75mm; Diam: 40 mm.
The container (lower part) is filled with 3M KCl solution.
The cover (upper part) has 4 ports to hold the current
and voltage electrodes. Electrodes can be connected
with wire and 2 terminal sockets for 1 mm pins.
Reference: AS-CONT
Price (container+cover): 185.-€
3 – Sockets and plugs (1mm).
Plug 1 mm Available in Black, Red, Blue and Yellow
Price: 0.95 € (all colors)
References:
Black: AS-PL-BK, Red: AS-PL-RD, Blue: AS-PL-BL, Yellow: AS-PL-YL
Socket 1 mm. Available in Black, Red, Blue and Yellow
Price: 2.45 € (all colors)
References:
Black: AS-SO-BK, Red: AS-SO-RD, Blue: AS-SO-BL, Yellow: AS-SO-YL
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Price list (in Euro)
Complete chamber assembly as depicted in Fig. 2A
6.5 mm version (price for 1 unit)
12 mm version (price for 1 unit)
Price reduction for order of 2 or more: 10%
2 complete assemblies of UC-65-CP
2 complete assemblies of UC-12-CP
Parts purchased separately
Filter specific parts
Main part 6.5 mm version (incl adapter ring)
Main part 12 mm version
Common parts
Chamber Support
Cover AP
Cover BL
Current electrode - complete assembly
Current electrode - Pt wire and pin
Current electrode - Lucite holder
Voltage electrode - complete assembly
Voltage electrode - Ag/AgCl pellet/contact
Voltage electrode - Holder
Inflow connection - adjustable length
Inflow connection - fixed length
Outflow connection
Closure of 6 mm port
Electric connection
Spare parts
Ferrule
O-ring - electrodes and in/outflow ports
O-ring - closure BL side
O-ring - seal around 12 mm Costar
O-ring - seal around 6.5 mm Costar
PE tubing for voltage electrode - agar bridge (30m)
Peek tubing - length 25cm
Tygon tubing - ID:1.3 - inflow connection (10m)
Tygon tubing - ID:2.4 - outflow connection (15m)
UC-65-CP
UC-12-CP
1684.00
1660.00
3030.00
2988.00
Reference
Unit price
MP-65
MP-12
672.00
645.00
CS
COV-AP
COV-BL
CE-AS
CE-PT
CE-HD
VE-AS
VE-AG
VE-HD
IF-ADJ
IF-FIX
OF
CL
VC-CON
170.00
180.00
140.00
172.00
95.00
85.00
203.00
95.00
126.00
89.00
44.00
55.00
30.00
8.00
FR-1
OR-1
OR-2
OR-3
OR-4
PE-30
PT-25
TGT-13
TGT-24
4.80
0.90
1.20
0.90
0.90
49.00
3.50
45.00
24.00
EP Devices Tervuurse steenweg 154, B-3060 Bertem (Belgium)
VAT BE 0733.058.494
TEL: +32-16-490.293
FAX: +32-16-700.177 E-mail: wvd-1@ep-devices.com