Cairn OptoSplit III User manual

www.cairn-research.co.uk
Free Phone: [phone] (UK only)
Tel: [phone]
Fax: [phone]

Important Information
Please Read Before Installing and
Operating Your OptoSplit III
For maximum reliability we recommend using the equipment within
certain guidelines. If in any doubt, then please feel free to contact our
technical support department who are always willing to help. (e-mail
[email protected]o.uk)
Note: Please do not adjust any control
before reading this manual.
The OptoSplit is supplied pre-aligned and centred, please refer to this
manual before making any adjustments.
When changing the Cairn Filter Cube, care should be taken not to
touch any optical surfaces. If any lenses, lters, or mirrors are marked
then they should be wiped with a clean lens tissue.

Contents
Introduction
Installation guide
Installing Filters into the Cairn Filter Cube
Installing Dichroic Mirrors into the Cairn Filter Cube
Inserting the Cairn Filter Cube into the Optopsplit III
Auxilliary Component Mounts
Installing the OptoSplit III in the Lightpath
Operation of the OptoSplit III
Adjusting and Locking the Aperture
Adjusting the Position of the Images
Focussing the Camera on the Aperture
Dual and Single Image Modes of Use
Locating the OptoSplit Images on the Camera
Centering Images on the Camera
Image Optimisation (Trim Adjustment)
Technical Summary
Technical support
1
2
2.1
2.2
2.3
2.4
2.5
3
3.1
3.2
3.3
3.4
3.5
3.6
3.7
4
5
1
2
2
3
3
5
5
6
7
7
10
11
12
15
16
17
18

Page 1
1 Introduction
In uorescence imaging applications it is often useful to acquire simul-
taneous images at multiple emission wavelengths. Traditionally such
applications have been restricted by the speed of an electronic lter
changer, or by the cost and complexity of adding extra detection hard-
ware. In many instances the region of interest (ROI) does not require
the full resolution of the camera, so the ideal solution would be to
simultaneously image at dierent wavelengths on the same camera
chip. In conjunction with a research microscope and a suitable CCD
camera the Cairn OptoSplit III allows the researcher to do exactly this.
The Cairn OptoSplit III is usually supplied with a rectangular aperture to
dene a ROI and includes controls to allow up to three images to be
positioned accurately and conveniently within the camera frame.
Images can be acquired using any imaging software and processed
either manually o-line or using an appropriate analysis tool such as
the Splitview module in Universal Imaging’s Meta series software or
Field Split in Andor Bioimaging’s iQ.
The instrument is usually congured to attach to the c-mount output
port of a research microscope, with a c-mount CCD camera tted to its
output. The design allows for connection to a variety of alternative
devices, so please consult with us if you intend using it in any other
conguration.
Installation of the OptoSplit III in the light path is straightforward and
does not require any special tools. Should you wish to make use of the
full input image without splitting (non-split mode), then exchanging
the triplecube for the bypass cube, and a simple adjustment of the
input diaphragm will permit this. We do however recommend removal
of the OptoSplit from the light path when the splitting mode is not
required to ensure the maximum light throughput to your detector.

Page 2
2 Installation Guide
2 .1 Installing Filters into the Cairn Filter Cube
Before using the OptoSplit III image splitter you will rst need to install
the appropriate set of lters and mirror for your application into the
Cairn Filter Cube. Instructions for these procedures are in sections 2.1
and 2.2. Always take extreme care when changing or adjusting the
lter and mirror sets to avoid damage or soiling. If you purchased your
OptoSplit III with a full lter set pre-installed, then you may wish to go
to section 2.4 as the following may not be pertenant.
The Cairn Filter Cube has spaces for
three 25mm lters. These are held in
place by locking rings which can be
removed using the tool provided. To
t a lter, remove the locking ring
and place the lter orientated so
that the arrow points into the Cairn
Filter Cube and towards the light
path. Then simply replace and
tighten the locking ring.
www.cairn-research.co.uk
www.cairn-research.co.uk
www.cairn-research.co.uk
Turn the locking
ring anticlockwise
to remove it.
Carefully place
the lter inside
the recess.
Replace the locking
ring and rotate it
clockwise to secure.
The arrow should be orientated
into the lter cube and towards
the light path.
Long Dichroic
(1st Position)
Short Dichroic
(2nd Position)
Short Wave FilterMiddle Wave Filter
Long
Wave
Filter
(View from bottom
of cube)

Page 3
In most applications, the dichroic beamsplitter(s) will have longpass
characteristics, so the longer wavelength of the lters should be
located in the straight through lter position, and the shorter
wavelength lters in the second/third position.
2 .2 Installing Dichroic Mirrors into the Cairn Filter Cube
To t your dichroic mirrors into the Cairn Filter Cube remove each of the
eight screws that secure the three sections together. You will then be
able to gently pull the cube apart. There is one steel locating rod in
each part of the cube to ensure the correct alignment when it is
reassembled. Once the sections of the cube are seperated remove the
dichroic holder and place the dichroic mirror(s) (active side down) into
the rectangular recess(es). Then simply replace the holder and
reassemble the cube and tighten the screws.
2.3 Inserting the Cairn Filter Cube into the OptoSplit III
The Cairn lter cube is designed to be easily accessible in order to
facilitate quick and easy changes of lter sets. Access to the lter cube
mount is gained by removing the access panel using the two handles
to pull gently and rmly. The lter cube mount will now be visible. The
mount attached to the rear of the Cairn Filter Cube is designed to mate
with the bracket on the internal wall of the OptoSplit III, so that the
small handle on top of the cube will be facing out. Once the Cairn Filter
Cube is mounted successfully, replace the access panel using the two
handles. (See over for diagram.)
Remove the 8 screws and
seperate the three sections. Remove the dichroic holders and
carefully place the mirror(s) in
the recess(es) active side down.
Re-assemble the cube
and replace the screws.

Page 4
Remove the access panel using the
two handles.
Mounting
bracket
Replace the access panel using the
two handles.
Mount the lter cube onto the brackect on the interior of the OptoSplit II.

Page 5
2.5 Installing the OptoSplit III in the Light Path
Before installing the OptoSplit III unit it is important to rst set up the
microscope, camera, and software to give a clear image of an object of
less than half the size the camera frame. Ideally this should be a real
sample with the appropriate optical properties for the installed lter
set. Failing this a standard bright eld image can be used, but this may
lead to arbitrary intensity dierences between the spectrally resolved
images.
First the CCD camera should be mounted on the
microscope C-mount output and the port adjusted
to give the sharpest possible image. Once a clear
image can be seen the camera should be switched
o and removed from the microscope.
The OptoSplit III should then be tted on to the
microscope with the diaphragm
orientated toward the output of
the microscope. The camera can
then be xed onto the output
port of the OptoSplit III, with the
top orientated so that the top of
the camera lies parallel with the
top of the OptoSplit III. If you are
tting onto a vertical mount then
the tops should still be orientated
in the same direction.*
2.4 Auxilliary Component
Mounts
To control the relative intensity of
the two pathways we recommend
using neutral density ltering in the
brighter pathway using the auxilliary
component mounts. This is required
only when there is a signicant
dierence in brightness between
the images. Auxilliary component mounts.
Top of camera
V1
V3 H3
V2 H2