TSS20 Ovodyne Electroporator

Operator’s Manual

Electroporation by
​
 OPERATING INSTRUCTIONS FOR
TSS20 OVODYNE ELECTROPORATOR

ABBOTSBURY ENGINEERING LTD

Unit 11 Stephenson Road
St Ives, Cambs, PE27 3WJ. England.
Tel: +44 (0)1480 468810
E-Mail:​[email protected]
VISIT OUR WEB SITE www.intracel.co.uk

1
 OPERATING INSTRUCTIONS FOR
TSS20 OVODYNE ELECTROPORATOR

​SAFETY WARNING
● The ​TSS20 Ovodyne​ is capable of delivering voltages of up to 100v at current levels of up to 100mA
depending on the voltage output. This can produce a violent electric shock and could damage or
destroy some biological preparations. Care must be taken when handling equipment connected to the
TSS20 outputs

● Always switch off the ​TSS20 ​before leaving it unattended.

● The ​TSS20 Ovodyne​ is intended for research purpose only. It is not intended for clinical use and its
outputs should not be connected to humans.

● In normal operation the current is limited to 30mA upto 60v and 2mA from 60-100v. Under ‘BOOST’
setting this can be overridden to deliver higher currents over the whole range for a single sequence.
Maximum available current under ‘BOOST’ will not exceed 100mA but will depend on the voltage
output setting (see SPECIFICATION).

Introduction
Electroporation is now a widely used technique for the transfer of genetic material into cells. The large variety of
cell types and applications have led to the development of sophisticated electroporation equipment offering broad
capabilities but often at a high price.

In the Mid 1990s it became evident that the chick embryo could provide a convenient, inexpensive and valuable
basis for genetic transfer work without involving live animal testing. Although largely empirical it was found that a
few low voltage d.c. square wave pulses delivered to electrodes across the area of interest of the embryo was
highly effective in achieving transfection.

The TSS20 has been continuously improved and refined to provide a simple inexpensive yet effective tool for us
in many areas of genetic and other research.

Description
The ​TSS20 Ovodyne​ is a square wave pulse generator with features specifically designed for use as an ​in ovo
electroporator. These include the ability to deliver single or a pre-programmed number of pulses with an
accurately defined duration and space between pulses. The microprocessor controlled circuit sampling at 4MHz
gives precise control of output pulses. The unit delivers a clean square wave with a voltage setting resolution of
0.1v.

Designed with operator safety in mind the TSS20 is suitable for use in teaching as well as research
environments. Automatic current limits are applied to output pulses exceeding 60 volts. These limits may be
over-ridden on a sequence by sequence basis. The maximum current output will not exceed 100mA.

There is a resistance measurement facility built in to the TSS20 Ovodyne that provides a simple means of
confirming continuity once the electrodes are in place across the subject to be electroporated. A resistance check
can ensure that reproducible electrical conditions are applied from subject to subject. Electrodes can become
coated after a period of use and resistance may noticeably increase. A resistance check may be used to ensure
that current limiting conditions are not encountered​.

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UNPACKING THE TSS20 OVODYNE
The TSS20 Ovodyne is supplied packed in a foam insert within a cardboard box. Contained in the box are:
● The electroporator
● Mains lead
● Instruction manual (this document)

If ordered at the same time, small accessories such as electrodes or electrode holders may also be contained in
the same box.

In the unlikely event of any deficiencies please immediately notify your supplier through whom the equipment was
obtained.

Should any damages have occurred in transit you should immediately contact the carrier and your supplier with
details.

Check that the power supply details shown on the back of the instrument correspond to your local
electricity supply.

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SPECIFICATION

Voltage range 0.1 - 99.9v

Voltage resolution 0.1v

Voltage accuracy ± 0.2v (3.0 - 99.9v)

Number of Pulses 1 - 999

Pulse Width 1 - 999ms

P.W. resolution 1ms

Space (Separation between pulses) 10 - 9990ms in steps of 10ms

Resistance measurement range 0 - 9998 Ohms

Resistance accuracy >10%

Current limit (normal) 2mA (60.1 - 99.9v)

30mA (0 - 60.0v)

Current limit (boost) 100mA (0 - 30.0v)

90mA (30.1 - 50.0v)
50mA (50.1 - 70.0v)
30mA (70.1 - 90.0v)
10mA (90.1 - 99.9v)

Isolation Output stage is floating and isolated to 3kV from mains and control circuit

Output connector 4mm sockets

Power Requirements 230v 50/60Hz

115v 50/60Hz

Trigger Start switch (front panel)

Footswitch (accessory)
Hand push button (accessory)

Weight 2.5Kg

Dimensions 80 x 235 x 22mm (H x W x D)

Please note: Abbotsbury Engineering reserve the right to change the specifications without notice

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CONTROLS

POWER Illuminated double pole mains switch. On (1); off (0 )

NOTE: On switch-off the last program is retained in memory (except delay
cancellation).

Adjust up. Increments selected parameter value. Progressively faster when held
down

Adjust up. Increments selected parameter value. Progressively faster when held
down

VOLTS Press to select

NO. OF PULSES Press to select
PULSE WIDTH Press to select
SPACE Press to select
RESISTANCE Press to measure actual resistance across output terminals / electrodes when
connected to preparation. Measured value is displayed for 5 seconds.

RESET ZERO Press to reset pulse counter to zero. Reset to zero before starting new
sequence.
NOTE: You can optionally cancel this requirement to reset. (See below).

START Press to send pulses (3 second delay) 3 2 1 appears at the right of the
sequence display after which the pulses are sent. The set pulses can also be sent
using the remote start switch or the footswitch accessory. A single 'bleep'
sounds on completion of the sequence.
NOTE: You can optionally cancel the 3 second delay to enable the first pulse to
be delivered instantly on activating ' start'. (See below).

STOP Press to stop a pulse sequence in progress. If a pulse sequence is interrupted the
number of pulses already sent is displayed. Pressing START again will deliver
a new set of pulses in the quantity originally set.

EXAMPLE:​ The unit is set to deliver 5 pulses but is stopped at 3. Pressing

start again will deliver 5 more pulses, the display ending at a total of 8.

The STOP· button may be pressed during the initial 3-second delay period. The
3-2-1 delay countdown will start again on pressing START.

NOTE: ​If the current limit is exceeded during a pulse sequence, the
sequence will stop (as if the stop button had been pressed) , the display will
show 'CURR' in the resistance window and 3 short ' bleeps' will sound.
You should therefore eliminate the reason for this (see OPERATION
parameter entry), press RESET and start the sequence again .

BOOST Press before sending a pulse sequence if you wish to override the 30mA current
limit in the range 0-60v or 2mA in the range 60.1 to 99.9v. Maximum 100mA is
made available depending on voltage output selected. See SPECIFICATION.

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OPTIONAL CANCELLATION
OF 3 SECOND DELAY AND
RESET REQUIREMENT This applies to all new TSS20 Ovodynes delivered after December
16th 2004. To de-activate the delay and reset requirement hold down
the' start' button (or footswitch, or handswitch) while you switch on
the 'power on' switch on the TSS20. A zero (0) will appear at the
right-hand end of the .display indicating that these features are
de-activated.

To re-activate the delay and the reset functions turn the mains power
switch off for 10 seconds and switch on again.

NOTE:​ If you switch off and on again too quickly you may lose
the indicator zero (0) on the right hand side and temporarily cause
the pulse sequence to function without the delay being
reintroduced. Simply switch off and leave for 10 seconds before
turning on again

.
OPERATION

Switching on:
Connect the mains lead to the TSS20 and switch on at the green illuminated power switch.

'Remote' switches:
If remote switch accessories are to be used, connect the push-button switch and / or the footswitch jacks to a
remote jack socket (front or rear).

Parameter entry:
Set up your electrodes in accordance with your planned electroporation protocol and connect them to the
output sockets. The red socket is positive relative to the black. If desired, check the resistance across the
electrode terminals whilst they are immersed in buffer to ensure that the system current limit will not be
exceeded.

EXAMPLE: ​Voltage setting 9, resistance measurement 900 Ohms

IE =​ Vc
R

Vc = Command voltage R = Measurement resistance

IE = Electroporation current
IE = 10mA (assuming electrode resistance is small)

If the current limit of 30mA is likely to be exceeded you may consider reducing the voltage increasing the
electrode separation or reducing the buffer conductivity. Alternatively you may use the BOOST control (see
below).

Parameter values may be set by pressing the light blue square on the membrane pad (a * appears against the
parameter selected) and then adjusting the value using the 'up' or 'down' arrow keys. Holding down the
arrow key will progressively speed the parameter change.

NOTE:​ Last program is retained in memory on switch-off (except delay cancellation). ​After
switching off allow 10 seconds before switching on again (otherwise the sequence counter may not
reset to zero).

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Use of the Boost Control
Should you wish to override the normal current limit you should press the BOOST key before sending the
pulse sequence. The sign 'BOOST' will flash in the resistance window. Once the pulse sequence has been
sent the boost is cancelled. To re-instate it press the BOOST key again.

Resistance Measurement
With the electrodes in position press the resistance button. A click is heard as the measuring device is
activated and the measured value is displayed for 5 seconds. The maximum measurable resistance is 9998
ohms. Over this value the resistance will display 'OVER' or - - - - . Electrode integrity may also be
checked by measurement at a known separation in saline solution or conductive buffer. Highly resistive
circuits are unlikely to result in current limiting conditions and therefore an over range indication for many
users will be unimportant.

Use of the resistance measurement facility may be necessary under low resistance conditions when using the
BOOST control (See 'use of boost control' above).

Some circuits exhibit a high resistance during measurement but under electroporation voltage this may break
down resulting in a far more conductive path forming between the electrodes. Under these circumstances the
current demand at a given voltage may be too great for the TSS20 even when using the BOOST control.
If reduction of voltage or buffer conductivity or increase in electrode separation is not possible you may need
to use the EP21 Current Amplifier accessory. This connects to the output of the TSS20. It delivers the pulse
parameters set on the TSS20 but allows currents up to l000mA to be drawn.
A separate operating manual is supplied with the EP21.
Use of the EP21 may extend the range of operation of the TSS20 allowing greater use of cuvettes and other
in vitro applications.

MAINTENANCE
No user serviceable parts are contained within the ​TSS20 Ovodyne​ and no routine maintenance is required.

​SAFETY WARNING
At no time should the cover of the TSS20 be removed other than by a suitably qualified technician. Before
removing the cover disconnect from the mains power supply.

Should the POWER switch fail to illuminate on switching on check that the mains plug is properly located in
the socket at the rear of the instrument. The TSS20 is protected from power surges by a 500mA quick blow
fuse (1A for 115v. versions) contained in a sliding compartment immediately beneath the input socket. The
compartment also contains a spare fuse. ​CAUTION: REPLACE A BLOWN FUSE ONLY WITH ONE
OF THE SAME TYPE AND RATING.

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ACCESSORIES
01-923-09 Remote push-button switch, cable and jack plug
01-920-09 Remote footswitch, cable and jack plug
01-92S-09 Adjustatrode electrode holder with cable and 4mm plugs
01-926-09 Single electrode holder with cable and red 4mm plug
01-927-09 Single electrode holder with cable and black 4mm plug
01-928-06 Insulated silver wire electrode (flattened pole) pk/2
01-929-06 Insulated Silver wire electrode with offset pole pk/2
01-931-06 Tungsten electrode, polymide insulated, with 10:1 tapered tip, Imm
exposure, with connector. pk/2
01-932-06 Tungsten electrode, as above but 2mm exposure pk/2
01-933-06 Tungsten electrode, as above, but 3mm exposure pk/2
01-934-06 Tungsten electrode, as above, but 4mm exposure pk/2
01-93S-06 Tungsten electrode, as above, but Smm exposure pk/2
01-700-06 Insulated platinum wire-tipped electrode on rigid stainless steel (0.8mm dia.)
shaft. Exposed 0.5mm dia. Pt wire tip 7.Smm. pk/2
01-708-06 Insulated platinum wire-tipped electrode on semi-rigid (0.8mm dia.)
silver wire shaft. Exposed 0.5mm dia. Pt wire tip 7.Smm. pk/2
01-918-02 EP21 Current Amplifier 230v SO/60Hz
01-918-03 EP21 Current Amplifier II5v SO/60Hz

Remote push-button and footswitches​. For convenience the TSS20 Ovodyne is fitted with jack sockets both
at the front and the rear. The hand push-button or the foot switch may be connected to either socket.

Electrode Holders. ​The body of the single electrode holder is 6mm o.d. x 100mm length stainless steel tube.
This houses a recessed 1 mm bore gold plated socket into which a suitable metal electrode may be inserted.
By ensuring that the metal electrode shank is slightly 'kinked' over the bare portion (approx. 2Smm) which
is inserted into the socket the electrode is held firmly and makes good electrical contact.

The ​Adjustatrode ​comprises two 100mm holder bodies linked by a flexible push-fit spacer block. Electrode
separation may be controlled by a knurled knob adjustment mechanism. Range of adjustment can be altered
by moving the spacer block up or down the shafts of the two holders.

Electrodes.​ Although you may have your own preference for electrode design and material lntracel provide
these main types shown above. The silver electrodes are made from 0.8mm diameter silver wire about
25mm of which is exposed for insertion into the holder. The slight bend or kink in the exposed portion
ensures a firm grip in the holder socket. About 40mm silicone rubber insulated electrode length protrudes
from the holder. The silver electrode poles are flattened to form a ' paddle' end approximately 0.8 x 2mm.
Silver electrodes are supplied with either a straight 'paddle' end or one which is offset to assist in positioning
around the part of the embryo being electroporated.

The tungsten electrodes are produced from 0.5mm diameter tungsten wire. The exposed portion for insertion
into the holder is bent slightly in a similar manner to the silver electrodes. The shaft protruding from the
holder is polymide insulated. The pole is tapered 10: 1 to form a rounded point. Due to the experimental and
often empirical nature of the technique tip exposure lengths of I 2 3 4 and 5mm are available to suit
individual preferences.

Platinum electrodes although relatively more expensive resist oxidation and coating during use, and can be
used repeatedly without cleaning between electroporations. Intracel's electrodes use 0.5mm diameter
platinum wire attached either to a rigid (0.8mm dia.) stainless steel tubular shaft or a semi-rigid (0.8mm dia.)
silver wire shaft. The silver wire shaft may be bent easily by the user for positioning. 7.5mm of platinum
wire is exposed below the silicone rubber shaft insulation. The end of the wire is cut at 45​o​ and the surface
abraded flat.

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WARRANTY

The ​TSS20 Ovodyne​ Electroporator is guaranteed against faulty workmanship and materials for one year from
date of despatch from Abbotsbury Engineering Ltd. If you have purchased the TSS20 from a distributor or dealer
warranty claims should be routed through this supplier.
Any warranty liability accepted by Abbotsbury Engineering Ltd shall not exceed Abbotsbury Engineering’s UK list
price for the instrument concerned, or Abbotsbury Engineering’s relevant International list price in the case of
purchases outside the UK, those prices being applicable at the time of purchase.

Before returning goods to Abbotsbury Engineering for repair it is essential to obtain a goods return number from
Abbotsbury Engineering.

Goods returned for service under warranty should be consigned freight pre-paid to Abbotsbury Engineering at the
address shown on the next page, packed carefully to avoid transit damage.

We make no charge for labour, materials, return carriage or freight for goods serviced under warranty. For goods
returned from overseas, however, we charge for customs clearance, import handling or document charges
including legalization if required on shipment back to the user.

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The TSS20 Ovodyne Electroporator is CE marked in compliance with
the European Low Voltage and EMC Directives

ABBOTSBURY ENGINEERING LTD

Unit 11 Stephenson Road
St Ives, Cambs, PE27 3WJ. England.
Tel: +44 (0)1480 468810
E-Mail:​[email protected]
VISIT OUR WEB SITE ​www.intracel.co.uk

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