Leica ANGLE TWO User Manual
Leica Angle Two
Computer-Assisted Stereotaxic System
User Manual
™
Living up to Life
Intended Use Statement
The Angle Two™ system is designed for stereotaxic surgery on small animals only and is not intended for use on humans.
Trademarks
Leica, Leica Microsystems and logo are trademarks of Leica Microsystems GmbH HRB 5187 and used under license.
Angle Two is a trademark of Leica Biosystems Richmond Inc. 36-2899171. Other trademarks are the property of their
owners.
Copyright
Leica Biosystems Richmond Inc. owns the copyright on this document and any associated software.
Leica Biosystems Richmond is part of the Leica Microsystems group of companies. Under law, our written permission is
required before either the documentation or the software is copied, reproduced, translated, or converted to electronic or
other machine-readable form, in whole or in part.
Doc. 95.8808 Rev A02
© Leica Biosystems Richmond Inc. 2010
Important Information for All Users
The term "Leica Microsystems" when used in text in this document refers to Leica Biosystems Richmond Inc.
Due to a policy of continuous improvement, Leica Microsystems reserves the right to change specifications without notice.
Warranty claims can be made only if the system has been used for the specified application and operated according to the
instructions in this document. Damage resulting from inappropriate handling and/or misuse of the product will invalidate the
warranty. Leica Microsystems cannot assume liability for any such damage.
Persons operating the Angle Two must be adequately trained and warned of any potential hazards or hazardous procedures
before operating the instrument. Only trained staff are to remove any parts from the instrument, and only if instructed within
this manual.
Repairs must only be carried out by qualified service personnel authorized by Leica Microsystems.
Safety
Safety notices in this manual are cautions. Cautions are notifications of hazards that could lead to damage to the instrument
or the animal undergoing surgery.
Cautions use symbols with a yellow border, as illustrated below:
Revision Record
Rev. Issued Sections affected Detail
A01 September 2009 All First release
A02 October 2010 Contact details, copyright, specifications Updated to Leica Biosystems Richmond Inc.
Contacting Leica Microsystems
For enquiries related to the Angle Two™ stereotaxic system:
Phone:
Email:
Web site:
+1 (800) 225 3035
+1 (815) 678 2000
myneurolab@leica-microsystems.com
www.myneurolab.com
For other Leica Microsystems products see the web site: www.leica-microsystems.com
Leica Angle Two™ User Manual Rev A02 © Leica Biosystems Richmond Inc. 2010 2
Contents
Leica Angle Two™ . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1
Intended Use Statement . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2
Trademarks . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2
Copyright. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2
Important Information for All Users . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .2
Safety . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2
Revision Record . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .2
Contacting Leica Microsystems. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2
1 Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 5
1.1 Basic Steps of Use . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 5
2 Instrument Installation. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 6
2.1 Checklist . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 6
2.2 Assembly . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 7
3 Components . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 9
3.1 U Frame . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 10
3.2 Manipulator: Linear Movements . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 13
3.3 Manipulator: Tilt, Rotation and Swing-Out . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 16
3.4 Probe Shaft . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 20
3.5 Vernier Scales . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 22
3.6 Computer . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 23
3.7 Ancillary Products . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 23
4 Software Overview . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 24
4.1 Atlas . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 25
4.2 Modes . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 26
4.3 Initial Settings. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 29
4.4 Coordinates . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 31
5 Alignment . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 33
5.1 Checklist . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 33
5.2 Procedure. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 34
6 Using the Instrument . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 42
7 Cleaning and Maintenance . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 47
8 Specifications . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 48
Software Licence Terms . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 50
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4
1
Introduction
The Leica Angle Two™ stereotaxic system is designed to perform surgery at any angle and position. Reaching the
target site is easier and more accurate than manual systems as the Angle Two is computer assisted – all
calculations are done for you – meaning no user errors and no pilot studies are needed.
Probe position is set with three linear movements (in anterior/posterior, medial/lateral and dorsal/ventral axes)
and two rotary movements (tilt and rotation). Encoders on the manipulator measure these positions and relay
them to the Angle Two software. Using this information the software calculates the exact distance the
manipulator must move to reach the target site at any tilt or rotation.
The Virtual Skull Flat
tilt. You don’t need to move the animal to align the skull horizontally or perform difficult calculations to
incorporate tilt into the target coordinates – the software does it for you, automatically setting appropriate
“distance to target” figures.
To use the Angle Two system choose one of the atlases available and then select the coronal plate that shows
the target site. Click on the screen to mark the site or alternatively use previously saved target coordinates.
Once the target site is entered move the probe to known reference points (typically bregma and lambda) which
the software uses to align the atlas plate with the animal’s brain. Then simply move the manipulator drives and
watch the AP, ML and DV displays count down to zero – indicating you have reached the target site.
™
feature in the software uses the bregma and lambda reference points to calculate skull
1.1 Basic Steps of Use
After unpacking and installing the instrument:
1. Align the instrument (see “Alignment” on page 33)
2. Select the target site (see “Selecting the Target Site” on page 43)
3. Position the animal (see “Positioning the Animal” on page 43)
4. Adjust the tilt and rotation of the manipulator (see “Manipulator: Tilt, Rotation and SwingOut” on page 16)
5. Locate the zero reference points (see “Locating Bregma and Lambda” on page 44)
6. Move the manipulator over the target site (see “Locating the Target Site” on page 45)
7. Drill and insert the probe (see “Probe Placement” on page 46).
Leica Angle Two™ User Manual Rev A02 © Leica Biosystems Richmond Inc. 2010 5
2
Instrument Installation
The Leica Angle Two™ system needs to be assembled before use. Use the information below to check you have
received all parts and to assemble the system.
2.1 Checklist
The list below has the standard items supplied in the Angle Two system. Additional or alternative
items may be included depending on the configuration ordered.
Angle Two instrument
U frame with 45° and 18° ear bars and species adaptor
One or two manipulators with one standard probe holder each
Stereotaxic Alignment Kit (dial indicator, T-bar, clamp and Allen wrench)
A brain atlas in book form appropriate to the species you have selected
‘Angle Two Support’ DVD
Computer (with Windows XP, Angle Two software v 3.0.0, and a rat and/or mouse brain atlas
installed)
Monitor
Keyboard
Mouse
Leica Angle Two™ User Manual Rev A02 © Leica Biosystems Richmond Inc. 2010 6
2.2 Assembly
When unpacked the Angle Two instrument has either one or two manipulators folded over the U
frame.
1. Unlock the tilt clamp and gently lift each manipulator upright into position.
The rotation encoder will point down and the tilt encoder to the left, away from the U frame.
Instrument Installation
Figure 1. Unlock the tilt clamp (highlighted) and gently lift the manipulator upright into position
Leica Angle Two™ User Manual Rev A02 © Leica Biosystems Richmond Inc. 2010 7
Instrument Installation
Figure 2. Angle Two cables and computer ports
2. Connect the five instrument cables into the ports at the rear of the computer. Each cable and
its corresponding port has the same label.
If you have a second manipulator, insert the cables into the second card provided.
3. Connect the mouse and keyboard into the computer’s USB ports.
4. Connect the monitor and computer power cables into an outlet.
5. Turn on the computer. The Angle Two icon is displayed on the desktop.
Figure 3. The Angle Two desktop shortcut
Caution
Do not pull or attempt to lift the instrument by any of its cables.
Caution
The computer must be connected to an earthed mains power outlet socket.
Leica Angle Two™ User Manual Rev A02 © Leica Biosystems Richmond Inc. 2010 8
3
Components
This chapter describes the parts of the Leica Angle Two
The components are divided into the following sections:
“U Frame” on page 10 – includes the ear bars, nose clamp and incisor bar.
“Manipulator: Linear Movements” on page 13 – the three coarse drives that move the manipulator in the AP,
ML and DV directions and the fine drive for small movements in the DV direction.
“Manipulator: Tilt, Rotation and Swing-Out” on page 16 – how to tilt and rotate the probe, and swing-out
for moving the probe away in order to get clear access to the animal.
“Probe Shaft” on page 20 – how to change the shaft’s vertical and horizontal positions and attach a probe.
system and their functions.
™
“Vernier Scales” on page 22 – how to use the vernier scales.
“Computer” on page 23 – lists the hardware and software provided.
“Ancillary Products” on page 23 – other products available from Leica Microsystems for stereotaxic surgery.
Leica Angle Two™ User Manual Rev A02 © Leica Biosystems Richmond Inc. 2010 9
3.1 U Frame
Incisor Bar
U Frame
Nose Clamp
Ear Bars
Incisor Bar
Vertical control
Horizontal
control
Figure 4 shows the Angle Two U frame with a rat adapter and 45° ear bars attached.
Some of the part names in the figures below link to sections of the manual where the part is
discussed in more detail.
Components
Figure 4. The Angle Two U frame
The animal is held in the instrument U frame with the head at the closed end of the ‘U’ and the tail
at the open end. The ear bars, nose clamp and incisor bar attached to the U frame secure the
animal during surgery.
Adapters for a range of species are available (see www.myneurolab.com).
Incisor Bar
The animal’s incisor teeth are placed over the incisor bar. The bar can be adjusted horizontally and
vertically using the thumbscrews indicated below.
Leica Angle Two™ User Manual Rev A02 © Leica Biosystems Richmond Inc. 2010 10
Figure 5. The incisor bar can be moved vertically and horizontally
Components
Nose clamp
Vertical control
Removes the nose
clamp
Caution
Do not use the rat incisor bar on a mouse. A mouse adaptor is available from Leica Microsystems
Note: Hamsters and gerbils can use the rat ear bars and incisor bar.
Nose Clamp
The nose clamp is placed over the animal’s nose and gently lowered using the thumbscrew
indicated. The nose clamp helps to keep the animal in position when injectable anesthesia is used.
Figure 6. The nose clamp
If using gas anesthesia with a nose cone, remove the nose clamp by undoing the thumbscrew and
insert the nose cone.
Ear Bars
The ear bars are tapered at one end to fit deep inside the ear canal. The instrument comes with
two rat ear bars (18° and 45°). Other animal ear bars and adaptors can be purchased from Leica
Microsystems.
After the bars are positioned in the animal’s ears, the animal is centered in the middle of the
U frame using the scales on each bar. Use this measurement to reproduce the exact position of the
animal for repeat studies.
Leica Angle Two™ User Manual Rev A02 © Leica Biosystems Richmond Inc. 2010 11
Components
The ear bars are locked into position using the locking levers. The right lever has a right-hand
thread, and the left lever a left-hand thread. The levers only need a quarter turn to lock the ear
bars.
Figure 7. Turn the lever to lock the ear bar into place
Caution
Use 45° ear bars if post-surgical testing is planned. Do not use 18° ear bars as they will break the
tympanic membrane of an animal.
Caution
Do not use the 45° or 18° ear bars on a mouse. The mouse’s breathing passages run between the
ears, and as the skull is very thin, the mouse can easily suffocate if too much pressure is applied by
the ear bars. The mouse ear canal is too small to admit the ear bars blunted to avoid damage in
rats.
Leica Angle Two™ User Manual Rev A02 © Leica Biosystems Richmond Inc. 2010 12
3.2 Manipulator: Linear Movements
The manipulator is attached to the U frame and is used to move the probe. The manipulator has
three linear movements:
AP (Anterior/Posterior): moves horizontally along the length of the instrument
ML (Medial/Lateral): moves horizontally across the width of the instrument
DV (Dorsal/Ventral): moves vertically towards the animal.
AP ML DV
Components
Figure 8. The three linear movements of the manipulator: AP, ML and DV
Leica Angle Two™ User Manual Rev A02 © Leica Biosystems Richmond Inc. 2010 13
Components
DV drive
ML drive
AP drive
Fine drive
Each linear movement is controlled by a coarse drive and, in the DV direction, by an additional fine
drive. When locating the target site, first use the ML and AP coarse drives to move the probe into
place above the target. Then lower the probe until it reaches the brain using the coarse DV drive.
Finally, engage the fine drive to continue DV movements to the target within the brain.
Figure 9. The AP, ML, DV and fine drives move the probe in linear movements
Caution
Do not continue to turn the drives at the end of their movement range, as it will damage the nut
below.
Leica Angle Two™ User Manual Rev A02 © Leica Biosystems Richmond Inc. 2010 14
Components
Fine Drive
The fine drive (see Figure 9) provides greater control and smoother movement in the DV direction
when placing or removing the probe from the brain. The fine drive helps prevent inserting the
probe too deep.
Using the Fine Drive
1. Push the fine drive towards the cog to engage it.
Figure 10. Engaging the fine drive Figure 11. Disengaging the fine drive
2. Turn the fine drive to move in the DV direction.
3. When finished, pull the drive away from the cog to disengage.
Note: When engaging the fine drive, if no click is heard, slowly turn the coarse DV drive until you
hear it click into place. Immediately stop moving the coarse DV drive.
Caution
Do not use the coarse DV drive when the fine drive is engaged, as it will damage the instrument.
Leica Angle Two™ User Manual Rev A02 © Leica Biosystems Richmond Inc. 2010 15
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