Eppendorf FemtoJet User Manual

FemtoJet

®

Operating Manual

Your local distributor:
www.eppendorf.com/worldwide

Eppendorf AG
22331 Hamburg · Germany
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Eppendorf North America, Inc.
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Westbury, N.Y. 11590-0207 · USA
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eppendorf® is a registered trademarkPrinted in Germany

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Contents

1 Safety precautions and application

2 Device description

2.1 Overview . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 6

2.2 Short description . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 8

2.3 Startup . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 10

3 Mode of operation

3.1 Display, variable regulators and keypad . . . . . . . . . . . . . . . . 12

3.1.1 Setting the injection parameters . . . . . . . . . . . . . . . . . . . . . . 12

3.1.2 Variable regulators. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 13

3.1.3 Keypad. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 14

3.2 Hand control / foot control . . . . . . . . . . . . . . . . . . . . . . . . . . . 17

3.3 Connecting the capillary holder . . . . . . . . . . . . . . . . . . . . . . . 18

3.4 Information on working practices. . . . . . . . . . . . . . . . . . . . . . 19

3.4.1 Compensation pressure p

3.4.2 Injection pressure pi. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 21

3.4.3 Injection time ti . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 22

3.4.4 Clean function . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 22

3.4.5 Manual or automatic injection . . . . . . . . . . . . . . . . . . . . . . . . 23

3.4.6 Constant pressure/constant working pressure . . . . . . . . . . . 23

3.4.7 General note . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 23

3.5 The first injection . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 24

3.6 Connecting an Eppendorf Micromanipulator . . . . . . . . . . . . . 25

3.7 Functions . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 26

3.7.1 Changing the capillaries . . . . . . . . . . . . . . . . . . . . . . . . . . . . 27

3.7.2 Setting the pressure unit . . . . . . . . . . . . . . . . . . . . . . . . . . . . 27

3.7.3 Switching on/off the loudspeaker. . . . . . . . . . . . . . . . . . . . . . 27

3.7.4 Switching on/off the background illumination of the display . 27

3.7.5 Constant pressure . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 28

3.7.6 Dewatering the pressure accumulator. . . . . . . . . . . . . . . . . . 28

. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 5

. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 12

. . . . . . . . . . . . . . . . . . . . . . . . . . 21

c

. . . . . . . . . . . . . . . . . . . 3

4 Cleaning and Service

4.1 Cleaning . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 29

4.2 Disinfection . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 29

4.3 Decontamination prior to shipping . . . . . . . . . . . . . . . . . . . . . 29

4.3 Service . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 29

No part of this publication may be reproduced without the prior permission of the copyright owner.

Copyright© 2006 by Eppendorf AG, Hamburg

. . . . . . . . . . . . . . . . . . . . . . . . . . . . . 29

B 5247 900.011-12/0108

Contents

5 Troubleshooting

5.1 Potential errors . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 30

5.1.1 Self-test . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 30

5.2 Warnings . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 32

5.3 Error messages. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 33

6 Technical data

7 Ordering information

7a Ordering information for North America

8 Index

Appendix A

A.1 Description of interface . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 49

A.2 Commands and queries . . . . . . . . . . . . . . . . . . . . . . . . . . . . 49

EC Conformity Declaration

. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 47

. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 49

Operation via a computer

. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 30

. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 40

. . . . . . . . . . . . . . . . . . . . . . . . . . . . . 41

. . . . . . . . . . . . . . .44

. . . . . . . . . . . . . . . . . . . . . . . . . . 49

2

1 Safety precautions and application

Observe operating instructions!

●

The device is only designed for use in the research lab!

●

●

The FemtoJet is a lab device. It may only be operated by appropriately
qualified lab personnel.

●

Only use the device for microinjection under observance of all relevant
legal regulations!

Before plugging in the device, please compare your voltage

●

requirements with the specifications on the identification plate!

The unit may not be connected to a socket that is not grounded!

●

Use the appropriate mains cable.

Electrical connections may be effected only when the device is

●

switched off!

●

The unit may be opened only by authorized service personnel!
Potentially lethal voltage inside the unit.

Do not use the unit if damaged. This applies in particular when the

●

mains cable is damaged.

If the unit is not used in accordance with purpose for which it is

●

intended, its protective function may be impaired.

●

●

In the case of independently-manufactured capillaries, ensure that the

●

outer diameter of the capillary corresponds to the specifications of the
capillary grip head.

●

In the case of extended periods of non-use, the unit should be
depressurized using the key and switched off before being
stored!

Caution!

Take care when mounting glass capillaries or tools!
Mounting must always take place without the use
of pressure! Capillaries which have been mounted
incorrectly may break loose under pressure. Never
point mounted capillaries at persons.

Caution!

Take care when using glass capillaries! During
mounting or when placed upon a hard surface, the
glass capillaries may break and glass splinters may
cause injury. It is therefore essential to wear eye
protection!

3

1 Safety precautions and application

Use the unit only for the described purpose! The unit and in particular

●

the high pressure of the Clean function may not be used for purposes
other than microinjection!

●

Transfer

If the device is passed on to someone else, please include the
instruction manual.

●

Disposal

In case the product is to be disposed of, the relevant legal regulations
are to be observed.

Information on the disposal of electrical and electronic devices in
the European Community

The disposal of electrical devices is regulated within the European
Community by national regulations based on EU Directive 2002/96/EC
on waste electrical and electronic equipment (WEEE).
According to these regulations, any devices supplied after 13.08.05 in
the business-to-business sphere, to which this product is assigned,
may no longer be disposed of in municipal or domestic waste.
They are marked with the following symbol to indicate this.

As disposal regulations within the EU may vary from country to
country, please contact your supplier if necessary.

●

Use only original Eppendorf accessories. Only original accessory
parts may be used. The only exceptions are auxiliary pieces of
equipment recommended by the manufacturer. Otherwise all claims
under guarantee and liability become void immediately. Use other
accessories only if you have a certificate from Eppendorf AG
confirming technical safety in use.

The responsibility for the correct functioning of the device passes

●

onto the owner or user if the device is maintained or serviced
improperly by persons not belonging to the Eppendorf service team or
if used for a purpose other than that intended. Eppendorf AG is not
liable for damage occurring from non-observance of these notes.
Guarantee and conditions of liability of the terms of sale and delivery
of Eppendorf AG are not expanded by the above notes.

●

The Eppendorf AG reserves the right to make technical alterations to
this device!

4

2 Device description

2 Device description

The FemtoJet enables minute quantities of liquid to be injected into cells.
The relevant parameters (injection pressure pi and injection time ti)

are set using the variable regulators. The key or connected hand or
foot control or the INJECT key of the connected micromanipulator are

used to trigger these two parameters. If no injections are being performed,
compensation pressure pc is applied permanently.

5

2 Device description

2.1 Overview

1

2

3

4

1

Display

– Displays relevant parameters during injection or setting

of parameters.

– Guides operator through functions.

2

Keypad

– To trigger actions.
– To call up the function menu.
– To switch to and from Standby mode.

3

Variable regulators

– To set parameters injection pressure pi, injection time ti and

compensation pressure pc.

– To select functions and parameters in Function mode.

4

Tube connection

– To connect the injection tube to the device via the bayonet joint.

6

2 Device description

Standard accessories:

1 Injection tube, 2 m
1 Universal capillary holder with grip head for capillary diameters
of 1.0 mm and adapter for Femtotips
1 Hand control for triggering injection and the Clean function
1 Femtotips, Femtotips II and Microloader
1 Mains cable
1 Operating manual

Optional accessories:

1 Foot control for triggering the Inject function
1 Connecting cable (Interface Cable 5171) to the Micromanipulator 5171
or InjectMan





7

2 Device description

2.2 Short description

Display

Display of all relevant parameters and information.

Function keys

Clean

To apply the maximum pressure for cleaning the capillary
to the tube outlet.

Auto / Manu

To switch between manual and automatic time-controlled
injection.

Count

To reset to zero the counter for injections which have been
actuated.

Inject

To execute automatic or manual injection.

Menu

To switch the device to menu control; selected functions
may be executed.

Standby

When the key is pressed briefly:
To switch the device between operating mode and standby
mode.
When the key is held down:
As above, but also deairs the pressure accumulator.

8

2 Device description

Variable regulator

In the normal operating mode, the parameters for injection are set by the
three variable regulators. The arrangement of the variable regulators
corresponds to the arrangement of the parameters in the two bottom lines
of the display:
From left to right: injection pressure pi, injection time ti and compensation
pressure p
In other operating modes, the variable regulators have either the
significance indicated in the display or no significance at all.

Tube connection

The injection tube is connected to the device via a bayonet joint.

.

c

9

2 Device description

2.3 Startup

Diagram of rear of device

– Electrical connection is effected via the mains socket with integrated

mains switch (1).

3
3

2

1

– Connection to an Eppendorf manipulator designed for injection is

effected via the 15-pin interface (2).

– Optional switches (e.g. foot control, hand control) or a computer can

be connected to the 9-pin interfaces.

Switching on the device

Set the mains switch on the rear of the device to "I". The unit is switched
on, the display lights up, the unit runs through a warm-up routine and is
then ready for operation. The injection mode is shown in the display.

The injection tube may not be connected when switching on and during
the initialization phase.

After being switched on, the unit runs through an initialization phase and
supply pressure is built up. This takes approximately three minutes.

10

2 Device description

The drainage valve of the pressure cylinder is switched several times at
the beginning of the initialization phase. The pressure cylinder is also drai­ned every 3 hours when switched on. This results in a certain noise level
following a series-connected reminder signal.

Switching off the device

If it is not going to be used for an extended period, the device should

be depressurized via the switch-off routine using the key. This
guarantees a long service life of the device (see Section 3.1.3).

The switch-off routine ensures that the system is dewatered and deaired
correctly. Do not disconnect the device from the mains supply until
dewatering and deairing have taken place. If only the mains switch is
actuated, the existing pressure remains in the system.

11

3 Mode of operation

3 Mode of operation

3.1 Display, variable regulators and keypad

Before operating the device for the first time, users should read this
section carefully. This section describes operation of the device only.
Handling of the injection capillaries is described in Section 3.3.

AUTO n= 0

pi= 150 hPa

pi[hPa] ti[s] pc[hPa]

150 0.2 50

AUTO n= 0

pi= 215 PSI

pi[PSI] ti[s] pc[PSI]

2.15 0.2 0.72

Display

The display shows the following:

1st line:
injection mode AUTO or MANU,
counter n for number of injections
performed

2nd line (bold):
The most-recently set parameter or
relevant parameter

Bottom lines:
Parameters which affect injection

Compensation pressure pc which is
applied permanently in the normal
mode is displayed as an actual
measured value.

The arrangement of the three
parameters shown, p
corresponds to the arrangement of
the variable regulators.

, ti, pc,

i

3.1.1 Setting the injection parameters

The injection parameters have to be adapted to the injection capillaries
used and the cell type and liquid which are to be injected.

The following injection parameters can be set using the variable
regulators:

Injection pressure pi 0; 5 – 6,000 hPa (0.00; 0.07 – 87.0 psi)
Injection time ti0.1 – 99.9 s
Compensation pressure pc0; 5 – 6,000 hPa (0,00; 0.07 – 87.0 psi)

The settings are in increments of 1 hPa/0.1 s. The display can show hPa
or psi, as required. The setting and internal pressure conversion are
always performed in hPa. Rounding for the display in psi can result in
increments of 0.01 to 0.02 psi.
The display of pressure in psi varies according to the size of the value
(e.g.: 9.99 may also appear in the display as 10.0).

12

3 Mode of operation

Slight deviations between the nominal value (specified value) and
displayed compensation pressure pc (actual value) may occur for
technical reasons. The relevant value is the actual value measured.

The following settings are used as a
basis for initial injection experiments
using Femtotip

pileft-hand variable regulator: 150 hPa
ti central variable regulator: 0.5 s
pcright-hand variable regulator: 50 hPa



/ Femtotip II:

Variable regulators:

Display:

AUTO n= 0

pc= 50 hPa

pi[hPa] ti[s] pc[hPa]

150 0.5 50

3.1.2 Variable regulators

The variable regulators are set so that when they are turned slowly, the
values are changed by 1 hPa / 0.1 second per notch (0.01 – 0.02 psi).
If the regulators are turned faster, the values are altered in larger
increments.

13

3 Mode of operation

3.1.3 Keypad

Inject

Performs automatic or manual injection.
Automatic injection is controlled via the
injection time ti, and the injection
pressure pi is used. In the second line,
injection time ti is counted down from the
nominal value.

Manual injection is performed for as long as the key is held down.
Injection pressure pi is used. In the second line, injection time ti is counted
upwards from 0.0.

If the FemtoJet is connected to the Eppendorf Micromanipulator
InjectMan NI 2, 5171 or Inject Man, the injection movement of the

micromanipulator is also started via the key. Both units then
combine as a system to perform semi-automatic microinjection.
Alternatively, the INJECT key of the connected micromanipulator or the

hand/foot control can be used.
If synchronization is set to "limit" (SYNC = LIMIT) in the INJECT function

of the micromanipulator, semi-automatic injection can only be triggered
via the micromanipulator.

Clean

The maximum available pressure for
cleaning is applied at the tube outlet for
as long as the key is held down. Once the
key is released, the pressure drops to
compensation pressure p
pressure curve is shown in the form of
a bar.

again. The

c

AUTO - INJECT n= 0

0.4 s

pi[hPa] ti[s] pc[hPa]

150 0.6 50

CLEAN n= 0

≡≡≡≡≡≡≡≡≡≡≡≡≡≡≡_____

pi[hPa] ti[s] pc[hPa]

150 0.2 50

14

3 Mode of operation

Auto / Manu

To switch between manual and
automatic, time-controlled injection.

In the case of manual injection, time
is counted upwards from 0.0 seconds.
The parameter t
bottom two lines in order to emphasize
that the time cannot be set via the
variable regulators in this case.

Count

To reset to zero the counter for injections
which have been actuated.

Menu

To switch the device to menu control;
selected functions may be executed.

is not displayed in the

i

MANU - INJECT n= 0

0.4 s

pi[hPa] pc[hPa]

150 50

In the first line, FUNC indicates that the
device is in the function menu and shows
which function number is active.
The second line contains the function
description (e.g. "Select pressure unit").

The third line requests an action (e.g.
press the key to change the
pressure unit). Current status: [hPa]

The two bottom lines show information
about which action is assigned to which
variable regulator (e.g. other functions
can be selected using left-hand variable
regulator p

).

i

FUNC 1
Select pressure unit
INJECT = PSI [hPa]

Tu r n pi to
select function

15

3 Mode of operation

Standby

in the case of a brief interruption

When key is pressed briefly:
The device switches to the Standby
mode

without

of the display switches off. "STANDBY"
appears in the display with the suffix
"Supply pressure = rxyz" as a code for
system pressure ps being maintained.

Application:
For temporary interruptions or use of

as a Reset function.

When key is pressed again:
The device is reactivated and runs
through a short warm-up routine.
This lasts approximately one minute.

Standby

with deairing of pressure accumulator

Hold down the key (approx. 2 seconds;
the display moves to "STANDBY"):

The device switches to the Standby
mode. The pressure accumulator is
completely deaired, the illumination of
the display switches off. "STANDBY"
appears in the display with the note
"Supply pressure = x" (x ~ 0).

A noise can be heard at this stage.

Application:
For correct shutdown, with deairing of the
entire pressure system in the event of
longer periods of non-use or standstill
(weekend or storage) with subsequent
switch-off using the mains switch.

Press the key again:
The device is reactivated and runs
through the warm-up routine.
This takes approximately three minutes.

deairing. The illumination

STANDBY

Supply pressure 6789

Hold standby

to exhaust

STANDBY

Supply pressure 0

16