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Integra Ti ™
USER MANUAL
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0120
Integra Ti™
Contents
Preface 1
Introduction to Integra Ti™ 2
Intended Use 2
Applicable Part Numbers 3
Microscope Compatibility 4
Integra Ti™ Safety Precautions 5
Guidance & Manufacturers Declarations 6
Safety Information / Symbols 9
Overview 10
Labelling 11
Specification Table 12
User Interface Icons 13
Installation - Summary 14
Installation - Unpacking 14-16
System Features 17
Installation - Nikon Microscopes 18
Nikon TE2000/Ti 18
Nikon TE200/300, D200/300 18
Installation - Nikon TMD 19-20
Installation - Olympus IX microscopes 21
Olympus IX51/71/81 21
Olympus IX50/70 21
2
Integra Ti™
3
Integra Ti™
preface
This manual provides all necessary information to use the Research Instruments
Integra Ti™ micromanipulation system.
The system should be operated by trained personnel only. All sections of this manual
should be read and understood fully before any operation of the system. If the
operator is unsure of any of the information contained in this manual they should
contact Research Instruments or an appointed representative before attempting
to use this equipment.
This manual is provided for the exclusive use of Research Instruments clients.
In no event does Research Instruments assume the liability for any technical or
editorial errors of commission, or omission; nor is Research Instruments liable
for direct, indirect, incidental, or consequential damages arising out of the use or
inability to use this manual.
© This manual is protected by copyright, all rights reserved, and no part hereof
may be photocopied or reproduced in any form without the prior written consent
of Research Instruments.
The Integra Ti™ manual belongs with the micromanipulation system and should be
passed on with the Integra Ti™ micromanipulation system if relocated to another
clinic
Integra Ti™ is a trademark of Research instruments Ltd. All other brand names are
trademarks of their respective owners.
1
Integra Ti™
Introduction to integra ti™
Intended Use
To precisely position microtools under a microscope whilst maintaining
sample temperature during assisted reproduction techniques (ART)
such as Intracytoplasmic Sperm Injection (ICSI) and Intracytoplasmic
Morphologically-selected Sperm Injection (IMSI)
Applicable indications for use are subject to the regulations of the country
into which the device is sold. Availability of a Integra Ti™ Micromanipula-
tor for clinical use is dependent on the regulatory approval status of the
Integra Ti™ Micromanipulator within the country the device is intended
to be sold into.
2
Integra Ti™
Introduction to integra ti™
Applicable Part Numbers
3
Integra Ti™
Introduction to integra ti™
Microscope compatibility
4
Integra Ti™
safety precautions
This indicates cautionary text which should be followed to avoid injury
to users or damage to samples.
ONLY use the power cable and power supply adaptor supplied with the
system. The cable to the power supply is the Disconnect Device for this
equipment. To remove all electrical power from this product,
disconnect the power cable from the electrical outlet. Equipment
should be positioned so as to allow easy access to the power cable. The
appliance coupler or mains plug is used as the disconnect and must
remain readily operable.
WARNING To avoid the risk of electric shock, this equipment must only
be connected to a supply mains with protective earth.
5
Integra Ti™
Guidance & Manufacturer’s Declaration
Guidance and Manufacturer’s Declaration
– Electromagnetic Emissions (IEC 60601-1-2:2007)
The Integra Ti™ is intended for use in the electromagnetic environment
specified below.
The customer or the user of the Integra Ti™ should assure that it is used in such
an environment.
Emissions test Compliance Electromagnetic environment -
guidance
Power 3.0 A/m 3.0 A/m Power frequency magnetic fields should
frequency 50/60Hz 50/60Hz be at levels characteristic of a typical
(50/60 Hz) location in a typical commercial or
magnetic field hospital environment.
IEC 61000-4-8
Note: UT is the a.c. mains voltage prior to application of the test level.
7
Integra Ti™
Guidance & Manufacturer’s Declaration
Note 1: At 80 MHz and 800 MHz, the higher frequency range applies.
Note 2: These guidelines may not apply in all situations. Electromagnetic propagation
is affected by absorption and reflection from structures, objects and people.
Note 3: These guidelines may not apply in all situations. Electromagnetic propagation is affected
by absorption and reflection from structures, objects and people.
a
Field strengths from fixed transmitters, such as base stations for radio (cellular/cordless)
telephones and land mobile radios, amateur radio, AM and FM radio broadcast and TV
broadcast cannot be predicted theoretically with accuracy. To assess the electromagnetic
environment due to fixed RF transmitters, an electromagnetic site survey should be considered.
If the measured field strength in the location in which the Integra Ti™ is used exceeds the
applicable RF compliance level above, the Integra Ti™ should be observed to verify normal
operation. If abnormal performance is observed, additional measures may be necessary, such
as re-orienting or relocating the Integra Ti™.
b
Over the frequency range 150 kHz to 80 MHz, field strengths should be less than [V 1 ] V/m.
8
Integra Ti™
Safety information / symbols
Symbol Meaning
9
Integra Ti™
Overview
Welcome to the installation, set-up and maintenance manual for the Research
Instruments Integra Ti™ Micromanipulation Workstation. This document will guide
you through all of the aspects of the workstation and assist you in the operation
of the Integra Ti™.
Figure 1
Integra Ti™ shown fitted on Nikon Ti microscope
Note: To prevent transit damage to the equipment, the manipulators and toolholders
are packed separately, and must be fitted before operating the instrument.
10
Integra Ti™
Labelling
Product Name
Serial Number
Year of Manufacture
Integra Ti™
SN / Follow
Instructions
for Use
QC
Research Instruments Limited
Bickland Ind. Park, Falmouth,
Cornwall, TR11 4TA, UK
0120 www.research-instruments.com
Quality Check
CE Marking Manufacturer’s Details
Figure 2
Power Rating
11
Integra Ti™
Specification Table
Part Description
X, Y and Z movement from one lever
Manipulator
Sub-micron resolution
- Fine Control
Adjustable X and Y travel, 5mm Z travel
X and Y movement
Manipulator
10 micron resolution
- Coarse Control
4mm X and Y travel
Fast pipette location
PL30 toolholders Single control pipette angle adjustment
- 15 to 40 degrees*
Metal insert as standard - optional ITO glass insert
Heated Plates Two additional independent heated plates built in
Can control optional external plate
LCD touch screen displays for temperature control and help
Displays
menu
Stainless steel stage plate
XYM stage
40mm travel in X and Y, 28mm per turn
Syringes Choice of SAS (air), SAS-SE (air) and SOS (oil)
12
Integra Ti™
User Interface icons
The following table explains the meaning of all the user interface icons required
to use and adjust the Integra 3™ settings and features.
Icon Meaning
Alarm sound on.
Low priority alarm. If alarm sounds finish procedure and investigate cause of alarm.
Exit screen.
Saves changes.
Cancels changes.
13
Integra Ti™
Installation - Summary
The following steps must be completed in order before the system is used.
1. The microscope should be fully assembled apart from the stage. The Integra
TiTM replaces the standard stage.
2. A 4x objective with spacer is supplied with the Integra TiTM as part of the
raised micropipette setup system. This objective should be fitted next to the
standard 4x objective.
3. Remove the Integra TiTM from its packaging and fit it to the microscope.
Ensure that the mains power supply is prevented from falling off the
bench as this is likely to cause damage to equipment.
14
Integra Ti™
Installation - Unpacking
Unpack the system, then remove the XYM transport bracket.
Figure 3
Undo these three screws and remove the
yellow transport bracket.
Tape
Figure 4
15
Integra Ti™
Installation - Unpacking
TDU5000 Micromanipulators
Releasing one TDU5000 at a time will make the task easier and will minimise the
risk of damage.
Figure 6
Hold the TDU5000 that you are releasing, and turn over the packaging bracket.
Remove the two screws that are holding that TDU5000 in place, using the green-
ended screwdriver.
Remove any additional foam or yellow coloured packing pieces from the
TDU5000. Repeat these steps to release the second TDU5000.
16
Integra Ti™
System Features
Primary heated Coarse x,y
stage insert control lever
Output pillar
Output pillar Secondary heated
(mounting for PL30
(mounting for PL30 stage insert
toolholder)
toolholder)
Figure 7
17
Integra Ti™
Installation - Nikon Microscopes
Completely assemble the microscope including the eyepiece tube and illumination
column. If the system is to be fitted to an existing microscope simply remove the
microscope stage before continuing with the installation.
Tilt the illumination column backwards. Carefully lower the Integra TiTM onto
the microscope. Once the stage is in place, secure it to the microscope with the
mounting screws provided.
NIKON TE2000/Ti
Figure 8
Figure 9
18
Integra Ti™
Installation - Nikon TMD
Completely assemble the microscope including the eyepiece tube and illumination
column. If the system is to be fitted to an existing microscope simply remove the
microscope stage before continuing with the installation.
First, assemble the front and rear mounting brackets. Note: the less curved parts
are the rear supports, the more curved parts are the front supports.
Hole towards
this side
Cutout corner
this side
More curved
parts
Less curved
parts Front bracket
Figure 11
20
Integra Ti™
Installation - Olympus IX microscopes
Completely assemble the microscope including the eyepiece tube and illumination
column. If the system is to be fitted to an existing microscope simply remove the
microscope stage before continuing with the installation.
To attach the Integra to the microscope first fit the two adaptors to the microscope
using the four screws provided. Tilt the illumination column backwards. Carefully
lower the Integra TiTM onto the microscope. Once the stage is in place, secure it
to the microscope with the mounting screws provided.
OLYMPUS IX51/71/81
OLYMPUS IX51/71/81
Figure 13
21
Integra Ti™
Installation - Olympus IX3 microscopes
Completely assemble the microscope including the eyepiece tube and illumination
column. If the system is to be fitted to an existing microscope simply remove the
microscope stage before continuing with the installation.
Before fitting the Integra TiTM, a plastic cover must be removed from the microscope.
This will not affect the operation of the microscope. Keep the cover and its
screws safe in case they need to be refitted in future. Fit the two adaptors to the
microscope using the four screws provided (as IX51/71/81 on previous page).
Tilt the illumination column backwards. Carefully lower the Integra TiTM onto
the microscope. Once the stage is in place, secure it to the microscope with the
mounting screws provided.
Figure 17.
Figure 15.
Remove
Screws
Figure 16.
22
Integra Ti™
Installation - Olympus IMT2
Completely assemble the microscope including the eyepiece tube and illumination
column. If the system is to be fitted to an existing microscope simply remove the
microscope stage before continuing with the installation.
Figure 19
23
Integra Ti™
Installation - Leica Microscopes
LEICA DMIRB, DMI3000/4000B/6000B
Completely assemble the microscope including the eyepiece tube and illumination
column. If the system is to be fitted to an existing microscope simply remove the
microscope stage before continuing with the installation.
To attach the IntegraTM to the microscope first fit the two adaptors to the microscope
using the four screws provided. Tilt the illumination column backwards. Carefully
lower the Integra TiTM onto the microscope. Once the stage is in place, secure it
to the microscope with the mounting screws provided.
Figure 21
Figure 22
DMIRB
DMI3/4/6000
24
Integra Ti™
Installation - Leica Microscopes
LEICA DMIL
2 x M8 screws Figure 23
2 x M3 screws
Figure 24
25
Integra Ti™
Installation - Leica Microscopes
LEICA DMIL
Fit the rear adaptor/
illumination column and the
front adaptor as shown, using
the M4 countersunk screws
provided.
Figure 26
Figure 27
26
Integra Ti™
Installation - Zeiss Axiovert 40
The original rear stage mount must be removed and replaced with the new one
supplied. If removed parts are stored safely then the microscope can be restored
to its original condition if necessary.
Figure 28
Figure 29
Figure 30
Pull the plate to the other side and
remove the remaining two screws.
Figure 31
27
Integra Ti™
Installation - Zeiss Axiovert 40
Figure 32
Figure 33
Figure 34
28
Integra Ti™
Installation - Zeiss Axiovert 40
Carefully lower the Integra TiTM onto the microscope. Once the stage is in place,
secure it to the microscope with the mounting screws provided.
Figure 35
29
Integra Ti™
Installation - Zeiss Axiovert 200/ Axio Observer
Completely assemble the microscope including the eyepiece tube and illumination
column. If the system is to be fitted to an existing microscope simply remove the
microscope stage before continuing with the installation.
Two objective spacers are provided for the second 4x objective. Most Axiovert 200s
use 0.8” threads (smaller spacer). Later Axiovert 200 and Axio Observer use M27
threads (larger spacer). Choose the spacer as appropriate to fit your microscope.
Carefully lower the Integra TiTM onto the microscope. Once the stage is in place,
secure it to the microscope with the mounting screws provided.
Figure 36
30
Integra Ti™
Installation - TDU5000 micromanipulators
Figure 37
V-block
TDU5000 INSTALLATION
Lift TDU5000 into position ensuring that the
location spigots on the sides of the TDU5000
locate in the V-blocks on the underside of
the Integra baseplate. Tighten the mounting
screw to secure in position.
Note: the left hand TDU5000 has the mounting
screw on the left hand side, and vice versa.
Figure 38
31
Integra Ti™
Installation - PL30 Toolholders
PL30 INSTALLATION
Slide the PL30 on to the output tube of the TDU5000. Do not tighten the rotation
lock screw at this stage. Repeat for the second PL30.
Output tube of
TDU5000
Figure 39
Figure 40
Do not adjust
this screw
32
Integra Ti™
Installation - Electrical
Power supply:
Plug the 12V 5A power supply into the socket as shown. If the system has been
supplied with a glass heated stage insert it will have an additional 24V or 12V
power supply. The two supplies have different connectors so they cannot be fitted
the wrong way around.
Note: the glass heated insert will not work without the second power supply. If
a metal heated insert is fitted the second power supply must be unplugged from
the stage.
Figure 41
Connect power
supply here
On/off switch
33
Integra Ti™
Installation - Micropipette Holder
Micropipette Holder Assembly (MPH)
The Integra Ti™ is designed to use industry standard 1.0mm diameter glass
micropipettes.
1. Thread the tubing through the MPH shaft and push firmly over the tube in the
MPH tip. Hold the tip and screw on the shaft.
Figure 42
4. Insert the micropipette into the MPH tip until it passes through the
O-ring and hits the stop (inside the MPH).
Insert Figure 45
5. Tighten the MPH tip to clamp the micropipette in place and complete
assembly.
Figure 46
MPH tip
34
Integra Ti™
Installation - Micropipette Holder
INSERTION INTO TOOLHOLDER
To insert the micropipette holder into the toolholder first raise the setup lever
located on the side of the toolholder (see page 38) and rotate the PL30 towards
the front if desired. Then support the underside of the toolholder with your thumb
or finger, lay the MPH across the slot in the toolholder and press evenly down until
the MPH clicks into position.
Removal From Toolholder
To remove the MPH raise the setup lever. Then gently lift the MPH out of the holder.
Figure 47
Tubing
Use only the hard polythene tubing supplied by RI. This is preferable to soft tubing
because it is much less easily forced off the MPH by fluid pressure, and because its
rigidity allows it to transmit volume changes from the air syringe to the micropipette
with greater accuracy.
35
Integra Ti™
Basic Operation
Coarse control lever left Coarse control lever right
On/Off switch
XYM y control
z
XYM x control y
x
36
Integra Ti™
Basic Operation
Rotating the fine control lever gives vertical movement over a range of 5mm.
Rotate the lever anticlockwise to move upwards, and clockwise to move
downwards.
The XYM y control (larger knob) moves the stage forwards and backwards. The x
control moves the stage from side to side. The stage can be centred by aligning
the hole in the circular insert with the end of the objective.
Avoid using the extremes of movement of the XYM when using
60x or higher magnification objectives. Objectives may contact the
mechanism as the nosepiece is turned.
REDUCTION
RANGE IDEAL FOR
RATIO
37
Integra Ti™
Initial Pipette Setup
REQUIRES THE USE OF TWO 4x ACHROMAT OBJECTIVES
(one fitted with a spacer)
Focus screw
access point
Pipette
Friction Travel screw
angle
screw
Travel locking screw
Rotation
lock screw Figure 50
Setup
lever
Axial drive
Pipette
rotator Figure 51
If desired, the friction screw can be used to make the toolholder stay in any
position between fully raised and fully lowered. Tighten the screw gently until
there is a little resistance. Move the setup lever and adjust the tightness of the
screw if necessary.
38
Integra Ti™
Initial Pipette Setup
Do not place your weight on the stage as this will affect the focus.
1. Move the fine (red) and coarse (blue stripe) levers to a vertical position.
Carefully rotate the fine control lever to the bottom of the movement and
then rotate twelve full turns upwards.
2. Take an empty and clean Petri dish (set up dish), and lightly scratch a cross in
its centre, on the upper surface. This must be the same type of dish as used
clinically.
3. Select a low magnification objective. Place the dish on the stage and use the
microscope focus control to focus on the cross. Do not adjust the microscope
focus again and do not remove the dish until pipette setup is complete.
4. Select the setup objective (4x with spacer).
5. Lift the toolholder levers (yellow stripe) to the raised position. Insert holding
and injection pipettes into the micropipette holders (MPH) and clip the MPHs
into the toolholders (PL30).
6. Adjust the pipette angle as required by turning the pipette angle knob.
7. Check that the bend is aligned by sighting along the micropipette holder and
pipette and turning the pipette rotator as necessary.
8. Use the axial drive to move the pipettes such that the tips are close to each
other.
9. Now look into the eyepieces. Rotate the whole PL30 to bring the tip of the
pipette to the centre of the field of view. Use the axial drive if necessary to
adjust the pipette tip sideways. The tip may be out of focus - this does not
matter and will be adjusted next. When the tip of the pipette is at the centre,
firmly tighten the rotation lock screw.
10. Adjust the focus screw, not the microscope focus, to bring the pipette into
focus. The focus screw is inside the PL30 and is reached by inserting a flat
screwdriver vertically into the hole shown on figure 39.
11. Once the pipettes are located correctly and in focus, select the low
magnification objective without spacer. The scratch on the Petri dish should
still be in focus.
12. Lower the toolholder levers fully. The pipettes should come into view slightly
39
Integra Ti™
Initial Pipette Setup
above the dish surface. If the pipette is too high or too low, the distance of
travel can be adjusted with the travel screw. Loosen the travel locking screw
and rotate the travel screw until the pipette is just above the surface of the
dish. Now gently tighten the travel locking screw.
13. Repeat the above steps to set up the injection pipette.
14. After setup the pipettes can be moved out of the way of the dish by raising
both setup levers and turning the PL30s to the rear. They can be clicked back
into the same position after the Petri dish has been placed on the stage.
Pipette with toe down Straight pipette Pipette with heel down
is correct for sperm is correct for is incorrect
immobilisation sperm injection for all procedures
40
Integra Ti™
Routine Pipette Setup
Do not place your weight on the stage as this will affect the focus.
If this procedure does not work at any stage (for example, if the pipette tip is a
long way out of focus at step 7), then carry out the initial pipette setup procedure
as described on the previous pages.
1. Move the fine (red) and coarse (blue stripe) levers to a vertical position.
2. Take an empty and clean Petri dish (set up dish), and lightly scratch a cross in
its centre, on the upper surface. This must be the same type of dish as used
clinically.
3. Select a low magnification objective. Place the dish on the stage and use the
microscope focus control to focus on the cross. Do not adjust the microscope
focus again and do not remove the dish until pipette setup is complete.
4. Select the setup objective (4x with spacer).
5. Lift the toolholder levers (yellow stripe) to the raised position. Insert holding
and injection pipettes into the micropipette holders (MPH) and clip the MPHs
into the toolholders (PL30).
6. Adjust the pipette angle as required by turning the pipette angle knob.
7. Bring each pipette into the centre of the field of view of the microscope by
using the axial drive knob (black). If the tip is blurred, rotate the fine control
lever slightly (not the microscope focus) to bring it into focus.
8. Align the tip of the pipette by turning the rotator on the end of the MPH as
necessary.
9. Once the pipettes are located correctly and in focus, select the low
magnification objective without spacer. The scratch on the Petri dish should
still be in focus.
10. Remove the setup dish and replace with your clinical dish. Lower the
toolholder lever fully. The pipette will come into view slightly above the dish
surface. Small height adjustments can be made by rotating the fine control
lever. If the pipettes are consistently too high or too low then carry out the
initial pipette set-up procedure.
41
Integra Ti™
Temperature Controller
When Integra TiTM is powered up it shows the product logo followed by the home
screen.
Figure 53
Figure 54
Alarm
If a plate is outside +/- 2.5oC from the set point before it has settled, an alarm is
triggered. Once the plate has settled it triggers an alarm if the plate is outside
+/-0.5oC from the set point. An alarm is also triggered if a temperature sensor is
not present or the plate is disconnected.
Icon Meaning
Medium Priority Alarm. Promptly finish procedure or return sample to incubator
before investigating the cause of the alarm
Low Priority Alarm. If alarm sounds finish procedure and investigate cause of alarm
Menu
Touch the Menu button on the operation screen for useful extra features.
Figure 55
ABOUT: Shows the serial number and the firmware version of the controller.
HELP: Tips for troubleshooting the device or for simple maintenance actions.
CONTACT: Research Instruments contact details.
BEEP IS ON/OFF: Enables or disables the audible overheating alarm.
RESET: Resets temperature setpoints and calibration on all plates to their default
values. A warning message as in figure 51 will ask for confirmation of the reset,
press “YES” to continue or “NO” to exit without saving changes.
EXIT: Returns to the operation screen.
42
Integra Ti™
Temperature Controller
Adjusting Specimen Temperature
The heated plates are factory calibrated such that when the controller is set to
37C then the surface temperature of the plate actually measures 37C.
The temperature inside a Petri dish will normally be slightly lower than the
heated surface. After the system has been installed in its operating location the
temperature of each heated plate should be adjusted to allow for the difference.
Change the set point if the temperature in the dish needs to be changed. For
example, the heated plate might need to be set to 38C in order for the specimen
to be at 37C.
Prepare a Petri dish with water and paraffin oil that mimics your normal dish
preparation and place it on the heated insert in its normal position.
We recommend using a probe thermometer calibrated at 37C, such as the RI IVF
Thermometer. Place the probe of the thermometer in the centre of the water drop
and allow the temperature reading to stabilise.
Figure 56
Touch the arrow buttons to increase
or decrease the set temperature.
Touch the SAVE button to save the
new value and return to normal
operation.
Figure 57
Allow a few minutes for the new temperature to settle and repeat if necessary.
Calibration
Calibrate only if the displayed temperature is different to the actual temperature
of the plate surface. Place the probe of a calibrated thermometer in good thermal
contact with the surface. Note: simply touching the probe on the surface is not
adequate. Use a purpose made surface probe or use thermal transfer paste.
Products sold for computer heatsinks are suitable, and RI can also supply suitable
materials.
43
Integra Ti™
Temperature Controller
After switching on, wait at least 10 minutes to allow the temperature to settle
before calibrating.
Figure 58
A warning message appears to
avoid accidental operation. Touch
YES to continue.
Figure 59
Touch the icon of the plate to
calibrate or CANCEL to go back to
the operation screen. If the plate
is not present then calibration is
disabled.
Figure 60
Touch the up or down arrows
to make the temperature on
the display the same as the
temperature shown on the
thermometer.
Figure 61
Touch SAVE to finish. A message will appear to ask for confirmation. Touch YES
to continue. Wait at least ten minutes for the temperature to settle. Check the
temperature on the thermometer before placing a dish on the surface.
MPH B
Figure 62
Controls are provided to bring the tips of two micropipettes (typically Zona Drilling
and Blastomere Biopsy) into close proximity to enable both to be used in one
operation.
MPH ‘A’ (closest to the PL30) should be set up first following the procedure on
page 38.
MPH ‘B’ should then be adjusted using the vertical and horizontal angle controls
and the axial drive knob to bring the tip of the micropipette into focus close to the
tip of the micropipette in MPH ‘A’. To avoid breaking the micropipette this should
be carried out with the setup lever raised and using the 4x objective with spacer.
45
Integra Ti™
SAS Screw-actuated Air Syringe
Assembly - DO NOT FILL THE SYRINGE OR TUBING WITH OIL
spigot 1. Turn the unit upside down.
2. Insert the end of the hard polythene
tubing through the hole in the side of
the base.
3. Ease the tubing over the spigot,
ensure that the tubing is securely
fitted.
Note: If the tubing becomes worn
at this connecting point it could be
a source of air leaks:- remove the
tubing, cut about 10mm from the
end, then reconnect the tubing.
Figure 63
4. The other end of the tubing is normally
hole hard polythene tubing fitted to a micropipette holder (see
page 24).
Operation
Suction/Injection is obtained by turning the rotator on top of the syringe.
Injection Suction
Equilibration button
(E-button)
Rotator
Figure 64
46
Integra Ti™
SAS Screw-actuated Air Syringe
Equilibration Procedure
This procedure balances the pressure in the syringe with the capillary action of
the pipette, which allows the most accurate control of sperm injection.
The following is a typical procedure. However, other methods are possible e.g.
using medium without PVP. Individual clinics may develop their own procedures.
E-button
Injection pipette:
Equilibrate with PVP.
Holding pipette:
Equilibrate with medium.
Figure 65
1. Place a drop of PVP/medium in the centre of a Petri dish.
2. Ensure the rotator is at the lower end of the movement. Turn the rotator down
(clockwise) if necessary.
3. Press the E-button (equilibrating button).
4. Using the toolholder setup lever, lower the micropipette into the drop.
5. Rotate the SAS anti-clockwise (i.e. upwards) 5 to 6 turns for an injection pipette,
or one turn for a holding pipette. Do not allow the PVP/medium to enter the
tubing. The level of the PVP/medium should always be clearly visible inside
the pipette.
6. Press the E-button.
7. Leave to equilibrate for 1-2 minutes.
Notes
If the sperm is moving in or out of the pipette too quickly, either the equilibration
is not complete or the rotator has been turned too far.
The E-button releases the internal air pressure when pressed. Capillary action will
cause the PVP/medium to move up the pipette. This can be countered with a small
clockwise movement of the rotator to balance the pressure.
47
Integra Ti™
SOS - Screw-actuated Oil Syringe
Overview
The SOS is a screw actuated syringe for the precise transfer and injection of
spermatozoa. By using oil (which has very low compressibility) to transfer the
movement of the control knob to the specimen, accurate aspiration and injection
can be easily achieved.
Cylinder
Piston
Figure 66
Fitting Tubing
Use hard polythene tubing as supplied by RI. Use of soft tubing will result in poor
specimen control.
Figure 67
Slide the nut over the tubing. Note: the non-threaded end should be pushed over
first.
49
Integra Ti™
SOS - Screw-actuated Oil Syringe
Push the end of the tubing firmly onto the tip of the fitting.
Figure 68
Tighten the nut onto the fitting. Use only finger pressure.
Figure 69
Fit the other end of the tubing to the pipette holder. See page 24 for details.
Figure 70
50
Integra Ti™
SOS - Screw-actuated Oil Syringe
Filling with Oil
RI recommends sterile filtered paraffin oil.
Figure 71
51
Integra Ti™
SOS - Screw-actuated Oil Syringe
Turn the SOS to a horizontal position. Add a few drops of oil to the fill port until
the oil is level with the end of the port.
Figure 73
Refit the cap ensuring that no air bubbles are trapped underneath. A small
amount of oil may be spilled, which should be wiped away with an absorbent
paper towel.
Figure 74
52
Integra Ti™
SOS - Screw-actuated Oil Syringe
With each procedure the
operating position of the
plunger will move further
towards the tip of the SOS.
When the plunger is close to
Figure 77 the tip as shown, the system
should be topped up with oil.
Figure 76
Next fill the port and replace the cap (see figures 55 and 56).
53
Integra Ti™
SOS - Screw-actuated Oil Syringe
Turn anti-clockwise to
aspirate.
Figure 78
Fitting a Pipette
Hold the pipette holder vertically and
turn the control knob clockwise until a
drop of oil emerges from the tip. Slide
the pipette into the holder and tighten
the tip.
Meniscus of oil.
Figure 79
54
Integra Ti™
Troubleshooting
Problem: Cannot focus onto the pipette tip
Answer: Are the pipettes heel-down?
The pipettes should be slightly toe-down. Adjust the angle on the PL30.
Problem: No fine z-movement (up and down) on micromanipulator.
Answer: Have you rotated the micromanipulator fine control lever too much,
either up or down, so that it has become locked?
The fine control Z movement has been rotated too much either up
or down. Rotate the fine control in the opposite direction to free the
locked movement.
If this does not work, lift off the secondary heated plate on the
affected side. Use the green ended driver from the toolkit to turn the
screw shown clockwise if the toolholder is high, anti-clockwise if the
toolholder is low.
Figure 80
Turn this way if the toolholder is high.
55
Integra Ti™
Troubleshooting
Problem: The stiffness of the fine or coarse lever is too tight or too loose.
Answer: For fine control levers. Select the green-ended hexagon driver provided
in your toolkit, loosen the locking screw shown below. Then adjust
the lever’s resistance by turning the adjuster anti-clockwise to loosen,
clockwise to tighten. Note: When adjusting the resistance, turn the
adjuster by the smallest possible amount. If any problems occur contact
the RI technical team.
Figure 81
Locking Adjuster
screw
Answer: For coarse control levers. Select the green-ended hexagon driver from
the toolkit supplied with your equipment.
Figure 82
Insert the driver into a screw head and slightly tighten or loosen the screw (less
than one turn is usually sufficient). Test the lever action to see if it is to your liking
(not too stiff, not too loose). If not sufficiently adjusted, insert the driver into a
different screw head and slightly tighten or loosen the screw. Note: the screws
must not be too loose or lost motion will be introduced.
56
Integra Ti™
Troubleshooting
Problem: Objectives hit stage insert when changing objectives or adjusting
focus.
Answer: Objectives are focussed too high.
Refocus on the top surface of a Petri dish.
Answer: The specimen is too close to the edge of the hole in the insert, causing
the objective to foul the side of the hole.
Position the specimen as close as possible to the centre of the hole in
the insert.
Answer: The Integra TiTM has been moved to a different microscope from that
originally specified.
The insert may need to be changed to suit the new microscope. Please
contact RI for more information.
Problem: The objectives hit the XYM stage mechanism.
The Integra TiTM has been moved to a different microscope from that
Answer:
originally specified.
The XYM stage may not be compatible with the new microscope.
Temperature controller
Problem: Stage insert or secondary heated insert too hot or too cold.
Answer: Insufficient settling time.
After switching on the system should be left for the display to show a
constant temperature before use.
Answer: Difference in temperature between specimen and plate surface.
Adjust set point to allow for the difference (page 43).
Answer: Temperature set point incorrect.
Touch the screen on the symbol corresponding to the affected insert.
Use the arrows to change if necessary then press OK.
Problem: Plate icon is grey.
Answer: The self-test can occasionally fail if the unit is switched on when still
warm. Touch the icon then click OK. Alternatively, switch off, allow to
cool, then switch on.
57
Integra Ti™
Troubleshooting
Injection
Problem: The specimen falls off the holding pipette / unable to get pressure
to inject.
Answer: Check that the tip of the MPH has been tightened properly. Tighten
the tip if necessary.
Answer: Is the micropipette tip blocked? If so, replace the micropipette.
Answer: There might be an air leak in the injection / aspiration system.
Check the connection of the tubing at each end. If it is loose, cut
approximately 10mm from the end of the tubing and reconnect.
Check the tubing for damage along its length, and replace if
necessary.
Replace the O-ring in the MPH tip (see page 59). If the O-ring is worn
out over a long period of time it will need replacement.
Replace the O-ring in the SAS air syringe (see page 60).
Problem: Poor control from syringes
Answer: SAS air syringe - check for
Air leaks as above
Blocked pipette
SOS oil syringe - check for:
Air bubbles
Blocked pipette
Use correct medium and oil e.g. Vitrolife ICSI™/OvoilTM
Oil leaks - treat as for air leaks
Problem: Unable to aspirate / inject?
Answer: Has the specimen dried up?
Warm stage heat can dry up an insufficiently protected specimen. A
generous layer of mineral oil over the specimen will protect it from
drying out.
Are the media you are using within their shelf-life?
Some media and PVP deteriorate if stored past their shelf life or if they
have not been kept refrigerated during storage or transit.
58
Integra Ti™
Maintenance
Manipulators
Figure 83
Body Tip O-ring
1. Unscrew the tip from the body of the MPH.
2. Remove the old O-ring. It may help to use a tool such as a small hexagon
wrench to push into the centre of the O-ring to remove it.
3. Take a new O-ring from your spares kit .
4. Place the new O-ring inside the body.
5. Screw the tip back onto the body of the MPH.
59
Integra Ti™
Maintenance
Air syringes
Inserting a New O-Ring : SAS-11/2-E
After a lot of use the O-ring may need replacing to ensure continuing smooth and
accurate operation. Spare O-rings are supplied with the syringe, and can also be
ordered from RI.
Unscrew the top to reveal the black rubber
O-ring. The O-ring is fitted in a groove in the
lower part of the syringe.
Carefully remove the O-ring.
Place the new O-ring in the groove.
Reassemble.
Top
O-ring
(in its groove)
SAS-SE Figure 84
The SAS-SE has been designed to reduce wear on the O-ring so that replacement
needs to be done less frequently. If you suspect that replacement is necessary
please contact RI as the replacement procedure is more complicated than for the
SAS 11/2-E.
60
Integra Ti™
Maintenance
Use a dust cover to protect the instrument when not in use. This will minimise the
need for cleaning. We recommend and supply a non PVC dust cover as plasticisers
commonly used in flexible PVC are known to be embryotoxic.
Always remove any spilt liquid or dirt immediately, i.e. keep instruments clean.
Cleaning
The Integra TiTM can be cleaned with a soft lint-free cloth moistened with isopropyl
alcohol only. This should be done with the instrument switched off at the wall
switch and the instrument should be completely dry before switching on. Never
use water to clean the Integra TiTM.
LIQUID SPILLS: If liquid is spilt inadvertently, IMMEDIATELY turn the electricity OFF
at the WALL SWITCH. Only then - quickly dry the instrument using paper towels or
similar. If it is suspected that liquids have gone inside the instrument then contact
RI for advice before switching on.
FUSES: ALWAYS replace a blown fuse with a CORRECTLY RATED new fuse.
Disposal of Goods
61
Integra Ti™
Repairs and Returns
RI REPAIRS SYSTEM
In the event that you have a problem with RI instruments, please follow the
procedure below to ensure rapid attention:-
First look at the ‘Troubleshooting’ section on the previous pages.
If you require any further help contact your distributor or RI direct. RI will try to
resolve the problem as quickly as possible.
RETURNS PROCEDURE
service@research-instruments.com
62