MIFE system requirements

MIFE system requirements - November 2007

NOTE: The MIFE system is being redeveloped during 2007 for greater functionality and user convenience and to run under Windows XP. The following are likely (some not yet certain) requirements for the new MIFE2 system.

Facility or component Source and cost

1 Electrophysiology laboratory with bench and Faraday cage Assumed present

There is need for general electrophysiological experience Assumed present or available

A vibration-isolated bench can be made from a strong table. On top of the table, a slab of steel or aluminium-covered composite is supported on three small, inflated inner tubes. All these components can be bought locally

2 Microelectrode preparation and filling facilities

Electrode puller. A simple one is sufficient. Those with multiple pulls can give lower-resistance electrodes. It is useful to be able to make tips of 2 to 5 mm diameter. This is standard in an electrophysiology lab. There are many suppliers.

Two simple (perhaps old) micromanipulators are needed for filling the electrodes. A background light is also needed. These are readily available or can be purchased.

A small oven, to 250 ° C, with gloves and electrode racks. It is in a well ventilated room or fume hood to disperse the silane safely. Readily available commercially.

Electrical test equipment: An adjustable mV source (0-50 mV, 0-500 mV).
Voltmeter (0.1 mV sensitivity) to measure the source voltage accurately. Usually present in electrophysiology labs. We have a diagram for a mV source.

3 Microscope with vertical or horizontal optic axis, for one of three configurations: A, B, C.

A. A standard compound microscope, lying on its back, is convenient for measuring fluxes from roots or other tissue immersed in solution. This configuration is illustrated with an E. nitens seedling. Long-working distance objectives and condenser are necessary. Electrodes are mounted on a multiple electrode mount (6 below), which itself is mounted on a manual manipulator (7 below) fixed above the microscope. The E. nitens illustration is of a manual manipulator that incorporates its own three electrode mount. The microscope stage is removed to provide space for a Perspex chamber to hold the tissue. The chamber is mounted on the driven micromanipulator (4 below). Thus, during flux measurements, it is the tissue chamber that actually moves relative to the electrodes. An existing microscope can be used.
Make the chamber to suit your tissue and microscope.

Your machine shop will do the mounting.

B. The configuration A could in principle be modified to have the chamber fixed (or on a manual manipulator) and the electrodes on their multiple mount mounted on the driven manipulator above the microscope. This configuration is generally more difficult to arrange mechanically. We have not used it.

C. An inverted microscope is good for single cell studies or protoplasts, particularly when patch clamping the same cell. The electrodes (if >1 are used) are on the multiple electrode mount (6 below) that is itself mounted on the driven manipulator in the usual configuration, as for patch clamping. Although the electrodes are at an angle of ~45^o to the horizontal, the mount can be mounted on a block to allow the driven manipulator axes, and electrode tip movement, to be horizontal and vertical. No multiple electrode mount yet exists that can be safely supported by the PatchMan NP2 (100 g load limit). The Marshauser SM325 (500 g load) is being investigated.

4 Driven micromanipulator

Eppendorf PatchMan NP2 (or equivalent). Marshauser SM325 is being investigated - decision by January 2008. These elegant stepper motor-based manipulators are controlled by joystick and by the MIFE2 software. Obtain a quote from your Eppendorf (or Marshauser) supplier.
Cost ~A$15,000.

The driven micromanipulator is mounted on a solid stand (obtainable from the supplier) or on a rigid support from the microscope body. Your machine shop may be needed to mount the micromanipulator.

5 Electrode holders and Ag/AgCl half cells
Commercial holders are available in various styles, with silver wires or with pellets. They must have a side hole for pressure relief as the prepared microelectrode is inserted. They should have a 1 mm hole for the electrical connector to the MIFE preamplifier.

WPI, Clark and others can supply them. See their catalogues.
For our own use, we have made our own holders.

6 Multiple electrode mount

A commercial 2, 3 or 4-electrode mount is needed if more that one ion is to be measured at one time. It is mounted on the driven manipulator as described above or on the 3-axis manual manipulator (7 below), of which it may be part. It must be able to hold the 6 mm diameter electrode holders, and have a sufficient angle ( ~ 22° ) between them so that the electrode tips can be adjusted together without their shanks touching. We are considering commissioning a 4-electrode mount. Cost is unknown, an estimate is A$5000.

7 Manual manipulator for adjusting the electrode position

A manual 3-axis manipulator is needed for general electrode positioning. It must have a long (>3 cm) axis parallel to the electrodes, to raise and lower them. We have used Narishige MM-3. The MLN-3 may also work. Other brands also will be suitable. See manipulator supplier’s web pages or catalogues. Median cost ~ A$3000.

The manipulator is mounted on a rigid stand or onto the body of the microscope, for optimum rigidity of support, in a way that depends on the configuration chosen. Mounting done by your machine shop.

8 PC computer

Any PC Windows XP will work as long as it has two USB ports. One data file can easily be >1 MB, so an archiving facility is desirable for effective data management. Your local supply.

9 Data acquisition module

The new MIFE2 software is written for the National Instruments USB6009 data acquisition module. We will supply this, mounted inside the MIFE amplifier box.

10 MIFE electronics hardware*

This is the central part of the MIFE system and is controlled by the software (which is included in the cost). Our estimated cost A$25,000*

11 The MIFE2/MIFEFLUX software*

This software is essential to control the hardware and to analyse the data. A handbook is provided. In the redeveloped MIFE2 system, the hardware will be controlled via a USB port to the USB6009. Included in the above cost for the hardware.

12 Spreadsheet software

General-purpose spreadsheet software is used to display and print the ASCII text files produced by MIFE2 and MIFEFLUX. Most labs will have this already

13 Consultancy, training and other MIFE set-up costs

The entire system to hold and view tissue, to prepare and set up electrodes, to adjust them close to the tissue and to move them to and fro, is technically complex. Freight and insurance: at cost. We can provide suitable consultancy services.

You may wish to have a consultant from Tasmania come to set up the MIFE system and give basic training in its use. The time required is likely to be about two weeks, but should be arranged by negotiation with UTAS Innovation Ltd. Consultants fee: to be negotiated; consultants travel: at cost; consultants accommodation and local expenses: at cost.

You may wish to visit the MIFE Centre for Membrane Transport at the University of Tasmania, in which the MIFE system has been developed, to be trained in its use and setting up. Time required is likely to be about two weeks. Suitable arrangements should be negotiated with UTAS Innovation Ltd. and Dr Ian Newman in the Biophysics lab. Hobart bench time: negotiated; your travel, accommodation and local expenses: at cost;

14 Guarantee and maintenance

The hardware is guaranteed against faulty workmanship for 12 months from delivery. Circuit diagrams will be provided and it may be more efficient for your local electronics technician to test and make repairs. You pay the freight to Hobart; the hardware will be repaired or replaced. We will pay the return freight.

No guarantee is given that the software will perform as expected. It is working well and is in regular use in the MIFE Centre, so it will be well maintained. Bugs will be corrected as quickly as possible. Improvements are made from time to time. If any problems are not solvable by studying the Handbook, please E-mail Ian Newman. Corrected and improved versions will be provided to all users without further charge.

* These items are made and supplied by the University of Tasmania (UTAS Innovation Ltd). Prices do not include Australian GSTax, and may change without notice. Please contact Dr Ian Newman, in the first instance, for a quotation for your particular MIFE requirements.

To MIFE Hardware page | Biophysics home page.

	Facility or component	Source and cost
1	Electrophysiology laboratory with bench and Faraday cage	Assumed present
	There is need for general electrophysiological experience	Assumed present or available
	A vibration-isolated bench can be made from a strong table. On top of the table, a slab of steel or aluminium-covered composite is supported on three small, inflated inner tubes.	All these components can be bought locally
2	Microelectrode preparation and filling facilities
	Electrode puller. A simple one is sufficient. Those with multiple pulls can give lower-resistance electrodes. It is useful to be able to make tips of 2 to 5 mm diameter.	This is standard in an electrophysiology lab. There are many suppliers.
	Two simple (perhaps old) micromanipulators are needed for filling the electrodes. A background light is also needed.	These are readily available or can be purchased.
	A small oven, to 250 ° C, with gloves and electrode racks. It is in a well ventilated room or fume hood to disperse the silane safely.	Readily available commercially.
	Electrical test equipment: An adjustable mV source (0-50 mV, 0-500 mV). Voltmeter (0.1 mV sensitivity) to measure the source voltage accurately.	Usually present in electrophysiology labs. We have a diagram for a mV source.
3	Microscope with vertical or horizontal optic axis, for one of three configurations: A, B, C.
	A. A standard compound microscope, lying on its back, is convenient for measuring fluxes from roots or other tissue immersed in solution. This configuration is illustrated with an E. nitens seedling. Long-working distance objectives and condenser are necessary. Electrodes are mounted on a multiple electrode mount (6 below), which itself is mounted on a manual manipulator (7 below) fixed above the microscope. The E. nitens illustration is of a manual manipulator that incorporates its own three electrode mount. The microscope stage is removed to provide space for a Perspex chamber to hold the tissue. The chamber is mounted on the driven micromanipulator (4 below). Thus, during flux measurements, it is the tissue chamber that actually moves relative to the electrodes.	An existing microscope can be used. Make the chamber to suit your tissue and microscope. Your machine shop will do the mounting.
	B. The configuration A could in principle be modified to have the chamber fixed (or on a manual manipulator) and the electrodes on their multiple mount mounted on the driven manipulator above the microscope. This configuration is generally more difficult to arrange mechanically. We have not used it.
	C. An inverted microscope is good for single cell studies or protoplasts, particularly when patch clamping the same cell. The electrodes (if >1 are used) are on the multiple electrode mount (6 below) that is itself mounted on the driven manipulator in the usual configuration, as for patch clamping. Although the electrodes are at an angle of ~45^o to the horizontal, the mount can be mounted on a block to allow the driven manipulator axes, and electrode tip movement, to be horizontal and vertical.	No multiple electrode mount yet exists that can be safely supported by the PatchMan NP2 (100 g load limit). The Marshauser SM325 (500 g load) is being investigated.
4	Driven micromanipulator
	Eppendorf PatchMan NP2 (or equivalent). Marshauser SM325 is being investigated - decision by January 2008. These elegant stepper motor-based manipulators are controlled by joystick and by the MIFE2 software.	Obtain a quote from your Eppendorf (or Marshauser) supplier. Cost ~A$15,000.
	The driven micromanipulator is mounted on a solid stand (obtainable from the supplier) or on a rigid support from the microscope body.	Your machine shop may be needed to mount the micromanipulator.
5	Electrode holders and Ag/AgCl half cells Commercial holders are available in various styles, with silver wires or with pellets. They must have a side hole for pressure relief as the prepared microelectrode is inserted. They should have a 1 mm hole for the electrical connector to the MIFE preamplifier.
5		WPI, Clark and others can supply them. See their catalogues. For our own use, we have made our own holders.
6	Multiple electrode mount
6	A commercial 2, 3 or 4-electrode mount is needed if more that one ion is to be measured at one time. It is mounted on the driven manipulator as described above or on the 3-axis manual manipulator (7 below), of which it may be part. It must be able to hold the 6 mm diameter electrode holders, and have a sufficient angle ( ~ 22° ) between them so that the electrode tips can be adjusted together without their shanks touching.	We are considering commissioning a 4-electrode mount. Cost is unknown, an estimate is A$5000.
7	Manual manipulator for adjusting the electrode position
	A manual 3-axis manipulator is needed for general electrode positioning. It must have a long (>3 cm) axis parallel to the electrodes, to raise and lower them. We have used Narishige MM-3. The MLN-3 may also work. Other brands also will be suitable.	See manipulator supplier’s web pages or catalogues. Median cost ~ A$3000.
	The manipulator is mounted on a rigid stand or onto the body of the microscope, for optimum rigidity of support, in a way that depends on the configuration chosen.	Mounting done by your machine shop.
8	PC computer
8	Any PC Windows XP will work as long as it has two USB ports. One data file can easily be >1 MB, so an archiving facility is desirable for effective data management.	Your local supply.
9	Data acquisition module
9	The new MIFE2 software is written for the National Instruments USB6009 data acquisition module.	We will supply this, mounted inside the MIFE amplifier box.
10	MIFE electronics hardware*
10	This is the central part of the MIFE system and is controlled by the software (which is included in the cost).	Our estimated cost A$25,000*
11	The MIFE2/MIFEFLUX software*
11	This software is essential to control the hardware and to analyse the data. A handbook is provided. In the redeveloped MIFE2 system, the hardware will be controlled via a USB port to the USB6009.	Included in the above cost for the hardware.
12	Spreadsheet software
12	General-purpose spreadsheet software is used to display and print the ASCII text files produced by MIFE2 and MIFEFLUX.	Most labs will have this already
13	Consultancy, training and other MIFE set-up costs
	The entire system to hold and view tissue, to prepare and set up electrodes, to adjust them close to the tissue and to move them to and fro, is technically complex.	Freight and insurance: at cost. We can provide suitable consultancy services.
	You may wish to have a consultant from Tasmania come to set up the MIFE system and give basic training in its use. The time required is likely to be about two weeks, but should be arranged by negotiation with UTAS Innovation Ltd.	Consultants fee: to be negotiated; consultants travel: at cost; consultants accommodation and local expenses: at cost.
	You may wish to visit the MIFE Centre for Membrane Transport at the University of Tasmania, in which the MIFE system has been developed, to be trained in its use and setting up. Time required is likely to be about two weeks. Suitable arrangements should be negotiated with UTAS Innovation Ltd. and Dr Ian Newman in the Biophysics lab.	Hobart bench time: negotiated; your travel, accommodation and local expenses: at cost;
14	Guarantee and maintenance
	The hardware is guaranteed against faulty workmanship for 12 months from delivery. Circuit diagrams will be provided and it may be more efficient for your local electronics technician to test and make repairs.	You pay the freight to Hobart; the hardware will be repaired or replaced. We will pay the return freight.
	No guarantee is given that the software will perform as expected. It is working well and is in regular use in the MIFE Centre, so it will be well maintained. Bugs will be corrected as quickly as possible. Improvements are made from time to time. If any problems are not solvable by studying the Handbook, please E-mail Ian Newman.	Corrected and improved versions will be provided to all users without further charge.