Rapid Solution Changers - RSC-160 & RSC-200 RSC injector Click to enlarge Unique features Fast and automated solution change No dead volume No cross contamination between solutions Precise setting of the flow with optional programmable motorized syringes Compatible with Axon and Heka software Introduction on RSC operation Change of concentration of ion, neurotransmitters or of all possible sort of ligands to be tested is an important issue in modern patch-clamp recordings. The Bio-Logic Rapid Solution Changers are the simplest and easiest-to-use systems to achieve this goal. These instruments use the principle known as "sewer pipes" in which the cell or the tip of the electrode (in excised-patch mode) is positioned close to the output of a capillary. The solution to be assayed is flowing out of the capillary at a moderate flow rate. The cell, being exposed in this stream, very rapidly equilibrates in the perfusion solution.  The fact that all solutions are flowing in individual tubes right to their end eliminates all problems of dead volumes, washing and cross-contaminations between solutions to be assayed (1,2). This perfusion principle has been used in many laboratories where it has been proved that it was a very adequate system for solution assays. Fast and automated solution change : The solutions to be assayed are driven to the rotating head of the RSC. The solution exchange is performed by a very precise rotation of the RSC head which exposes the patch pipette or the cell to the flow of one of the tubes. Times of rotation from one tube to the adjacent one can be programmed from 2 to 800 ms. The motor is light and vibration free thanks to the disk rotor technology and to the micro step command delivered by the controller. Due to the high number of tubes it is possible to have one tube for every solution to be assayed. As compared to fixed tube devices, the rotating head permits a fast change of the perfusion solution around the patch pipette without any contamination from the previous solution at the time of solution switching. Flow setting : Two different possibilities are available for the setting of the flow: gravity (The flow of other solutions can be stopped with the optional Valve Array or the solutions can flow simultaneously in the bath outside the range of the pipette). The solutions are simply driven by gravity and the flow rate controlled by the height of the reservoir containing the solutions.  optional motorized syringe array(s) . Each solution is contained in a glass syringe driven by its own stepping motor. The flow is adjustable and the start/stop can be synchronized with the movement of the RSC head. Control of the instrument : Solution changes and valve/motorized syringe control may be performed: Manually from the front panel of the controller. Automatically using analog pulses generated from a stimulator or a D/A board. (see for example : AN#1 - RSC - Control through Axon software ; AN#2 - RSC - Control through Heka software) Automatically using a PC computer linked via RS-232 or GPIB (RSC-200 only - NI GPIB boards only) and the provided software to edit and download a sequence. The RSC-200 doubles as a stimulator since it can deliver an analog stimulation to the patch-clamp amplifier through a 16-bit D/A converter. This stimulator can be programmed in combination with the solution change by the software. Description of RSC components Click to enlarge The RSC-160 & RSC-200 controller Front panel functions TUBE : tube selection SPEED : speed of rotation selection (2ms to 800ms) REF.: reference tube selection SETUP : set-up informations: head model, control mode, EV OPEN : open electrovalve/start programmable motorized syringe EV CLOSE : closed electrovalve/stop programmable motorized syringe VHOLDV : hold potential set-up (RSC-200 only) REM./LOC : remote or local control START, STOP : start and stop of sequence execution STATUS : functions status Rear panel functions ANALOG OUT (RSC-200 only) : stimulation output, +/-10V range; 1mV, 1ms step ANALOG CONTROL-IN : a signal sent to this input can drive the rotating head and the valve opening. In this mode only the valve in the active position will be opened. When the signal is detected, the corresponding valve is opened. After a programmable delay [0..200ms], the tube is moved to its corresponding position and the previous valve is closed: 0mV: no effect 100mV: valve #1, tube #1 200mV: valve #2, tube #2 ... 2000mV: valve #20, tube #20, etc. ANALOG CONTROL-OUT: this output reflects the current tube number and can be recorded, for example, together with a patch signal. tube #1: 100mV tube #2: 200mV tube #3: 300mV TRIGGER IN: A rising TTL edge on this input can be used to synchronize with an external signal. TRIGGER OUT: At this output 4 TTL signals can be controlled by the software ELECTROVALVES: Connection for the valve/motorized syringe sets DTR REMOTE: Remote control output for Bio-Logic DTR recorders RS-232: Serial port connection GPIB: GPIB port (RSC-200 only - NI GPIB boards only) MOTOR: Motor connection Electrovalves The electro valves are an optional device which opens the flow only on the selected tubes. If valves are not used all solutions would flow simultaneously, as in the RSC-100 instrument. The set of 9 valves are mounted into their housing. The RSC-160 system can drive up to 9 valves (1 set of valves) while the RSC-200 system can drive up to 36 valves (4 sets of valves). The valves used for RSC-160 & RSC-200 are normally closed pinch valves. The valves can be commanded by front panel of the controller or the software. All valves can be programmed to open or close synchronously with a tube position change. More than one valve can be opened at a time, or valves can be opened or closed independently of the head rotation. For example to pre-establish the flow in the next tube to be tested a few seconds before rotation. Motorized syringes   Click to enlarge 3d view of the MS-4 The motorized syringes have been developed to give to the user a possibility of flow control and sample economy. The motorized syringes are delivered in modules of 1, 4 or 8. Each syringe has an individual stepping-motor. The motor speed (flow rate) is adjusted (one knob per module). The stops and starts of the motors can be activated manually or by RSC or MSC commands (like the electrovalves). Electrovalves and motorized syringes can be used alternatively or in parallel. Syringe Rack and Rack Holder The most convenient way to hold the reservoir syringes is to use the syringe rack that has been especially designed for this use. The syringe rack can be installed or hanged at the most convenient location in the Faraday cage or set on its holder . One syringe rack can be set on each side of the rack holder, giving a total capacity of 18 syringe reservoirs. The Heads One of the important features of RSC heads is an easy exchange of solution delivering tubing, this eliminates necessity of washing and risks of residual contaminations. Every RSC-160 & RSC-200 system is delivered with one head. The straight head was designed to carry standard capillary glass, OD 1.00mm. This head gives the following advantages: Capillaries can protrude, giving better approach to a cell. The number of capillaries installed is up to the user. Capillaries take minimum space in the cuvette. Easy exchange of capillaries. Easy attachment to the flexible tubing Click to enlarge The head is made of Delrin, reinforced by an internal metal rod, for best shape stability. The outside diameter of the head is manufactured with the highest precision (+/- 10 microns) to give a good reproducibility of the tube positions. The head is equipped with two guiding pins for straight and parallel positioning of capillaries. The capillaries are kept in their position by two O-rings of precise diameter and tightened with a thermo-retractable sleeve. The capillaries are connected to a short piece of polyethylene tubing, which is connected to larger tubing going through electro valves into the reservoir syringes. Glass capillaries The straight head is designed to work with glass capillaries of an outside diameter 1.00mm (Ref: GB 100TF 8P - thin wall with filament, ID=0.78 mm, length: 8cm, fire polished ends). Other glass brands can also be used. It is recommended to check the OD of the selected capillaries with a caliper to select capillaries with OD close as possible to the theoretical value. Note: Other glass diameters can also be used with RSC-160 & RSC-200 only. In case of other diameter, the "step value" must be adjusted in the set-up menu of RSC-160 & RSC-200. The RSC-100 has no "step value" adjustment possibility. Flexible tubing To connect reservoir syringes with capillaries installed on the straight head, two types of tubing are used in standard version (see below for high flow rate set-up). 1) Polyethylene tubing (ID: 0.3, OD: 0.70mm, Reference: MERCK CLEVENOT BIE-03401-0001) is inserted into the rear end of the capillary tube. Approximatly 10-15cm length is needed. This tubing is used to interface with bigger diameter tubing coming from reservoir syringes and to minimise the load on the rotating head. 2) C-Flex tubing (ID: 0.5mm, OD: 2.1mm, Reference: COLE PARMER #6424-59 or BIOBLOCK SCIENTIFIC #M71195) is used to connect the reservoir syringe with the polyethylene tubing coming out of the capillary glass. C-Flex tubing is well adapted to our electrovalves and is much less permeable to air than, for example, silicon tubing. However other types of tubing can also be used. Stimulator The RSC-200 is equipped with one stimulator channel (+/-10V, 1mV and 1ms resolution). This stimulation is programmed by the same software as the solution change. TTL On the rear panel of the instrument 4 x TTL , independently programmable channels are available. As for the stimulation channel, the TTL pulses are programmed by the same software as the solution change. Software The RSC-160 & RSC-200 instrument are delivered with a Windows 3.1 software (Windows 95/98 compatible). The software permits the following operations: manual positioning of the head manual opening of the valves resting Vhold setting edit and download of a sequence where the following parameters can be exchanged at each step: duration of stay in the active position for a given tube number speed of solution change (head rotation) choice of electrovalves to be opened/motorized syringes to start (if applicable) stimulation pulse level ( range: +/-10V) (RSC-200 only) TTL signal activation (4 lines of signal available) triggering of other auxiliary instruments Computer requirements IBM-PC-AT or compatible with 486DX33 or higher. 4 Mbytes of RAM or more. VGA Monitor and VGA graphic card (640 x 480) or better. Computer connection via serial port or GPIB (RSC-200 only - NI GPIB boards only) Windows 3.1 or higher. Manipulator Click to enlarge   The RSC-160 & RSC-200 motor can be mounted, on any 3-axis manual manipulator, providing that the motor will have a mechanically stable fixation. We recommend 3 models of manipulators; UM-3C (with rotation mechanism) (You), M-152 (Narishige), MM-33 (Marzhauser) with direct mounting (no adaptor required - better mechanical link). These manipulators may be delivered with magnetic stands. Another very good solution for the micromanipulator fixture is the use of the Bio-Logic PCS-1000 Patch-Clamp-Stand. Click to enlarge   RSC-160 & RSC-200 Stepping motor II bottom view. The four central holes are used for micromanipulator mounting. All dimensions are in millimeters. Comparison between RSC-160 & RSC-200   RSC-160 RSC-200 Tube control up to 9 up to 36 Valve or motorized syringes control (with optional EVH-9 or MS-x Unit(s)) up to 9 (with 1 x EVH-9 or MS-1 + MS-8) up to 36 (with 4 x EVH-9 or 4 x [MS-1 + MS-8]) Built-in stimulator No Yes GPIB interface No Yes All other specifications are the same for the two instruments Ordering information The RSC-160 & RSC-200 are both delivered complete with: command box, stepping motor, driving software, one syringe rack and one "Straight head" (and a tubing kit). All other parts listed below are optional and should be specified on order or may be purchased separately. The straight head is delivered with a tubing kit (Ref: 025-00/18) composed of: 100 x Glass capillaries of an outside diameter 1.00mm (Ref: GB 100TF 8P - thin wall with filament, ID=0.78 mm, length: 8cm, fire polished ends). Polyethylene tubing (5 meter length, ID: 0.3, OD: 0.70mm, Reference: MERCK CLEVENOT BIE-03401-0001). C-Flex tubing (7,5 meter length, ID: 0.5mm, OD: 2.1mm, Reference: BIOBLOCK SCIENTIFIC #M71195). 9 x luer lock female. 9 x 10ml plastic syringe. 9 x plastic manual valve. 2 x O-Rings. Reference Description 025-40/A or /B RSC-160 Rapid Solution Changer 220V or 110V 025-50/A2 or /B2 RSC-200 Rapid Solution Changer 220V or 110V Manipulators     Options 025-00/13 EVH-9 9-Electrovalve Unit 025-00/24 MS-1 MOTORIZED SYRINGE FOR RSC/MSC 025-00/25 MS-4 MOTORIZED 4 SYRINGES FOR RSC/MSC 025-00/26 MS-8 MOTORIZED 8 SYRINGES FOR RSC/MSC 025-00/06 Syringe rack holder (1 or 2 RACK) 025-00/05 Additional Syringe rack (for up to 9 x 10ml syringe) Consumable 025-00/18 Tubing kit for straight rotating head

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