Cairn OptoSplit II Manuel utilisateur
www.cairn-research.co.uk
Free Phone: 08453301267 (UK only)
Tel: +44 (0) 1795590140
Fax: +44 (0) 1795594510
Important Information
Please Read Before Installing and
Operating Your Optosplit II
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 II
Auxilliary Component Mounts
Installing the Optosplit II in the Lightpath
Operation of the Optosplit II
Adjusting and Locking the Aperture
Adjusting the Position of the Images
Focussing the Camera on the Aperture
Centering Images on the Camera
Image Optimisation (Trim Adjustment)
Single Channel (non-split) mode
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
4
5
1
2
2
3
3
5
5
6
7
7
9
10
11
12
13
15
1 Introduction
In uorescence imaging applications it is often useful to acquire simul-
taneous images at two emission wavelengths. Traditionally such appli-
cations have been restricted by the speed of an electronic lter
changer, or by the cost and complexity of adding a second camera to a
system. 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 two dierent wavelengths on the same
camera chip. In conjunction with a research microscope and a suitable
CCD camera the Cairn Optosplit II allows the researcher to do exactly
this.
The Cairn Optosplit II is usually supplied with a rectangular aperture to
dene a ROI and includes controls to allow the two 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 congured 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
conguration.
Page 1
2 Installation Guide
Before using the Optosplit II 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 II with a full lter set pre-installed then you may wish to go to
section 2.4. as the following may not be pertenant.
2 .1 Installing Filters into the Cairn Filter Cube
The Cairn Filter Cube has spaces for
two 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.
In most applications the dichroic beamsplitter will have longpass char-
acteristics, so the longer wavelength of the two lters should be
located in the straight through lter position, and the shorter wave-
length lter in the second position. (Please refer to the technical sum-
mary in section 4 for details of the light path.)
CAIRN
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CAIRN
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CAIRN
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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.
Page 2
2 .2 Installing Dichroic Mirrors into the Cairn Filter Cube
To t your dichroic mirror into the Cairn Filter Cube remove each of the
four screws that secure the two halves together. You will then be able
to gently pull the two halves apart. There is one steel locating rod in
each half of the cube to ensure the correct alignment when it is
reassembled. Once the two halves of the cube are seperated place the
dichroic mirror (active side down) into the rectangular recess. Then
simply reassemble the cube and tighten the screws.
2.3 Inserting the Cairn Filter Cube into the Optosplit II
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 II 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 4 screws and
seperate the two halves. Replace the top half and
resecure the screws.
Place the dichroic holder in
the recess
Remove the dichroic holder
Page 3
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 4
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 one image is
disproportionately brighter than the
other.
This auxilliary position can also be used for corrector lenses to reduce
chromatic abberation and ensure both pathways are focussed on the
camera sensor.
2.5 Installing the Optosplit II in the Light Path
Before installing the Optosplit II 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 dierences between the spectrally resolved
images.
Firstly 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 II 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 II, with the top orientated
so that the top of the camera lies
parallel with the top of the Optosplit
II. If you are tting onto a vertical
mount then the tops should still be
orientated in the same direction.*
V1
V2
H2
Auxilliary component mounts.
Top of camera
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OPTOSPLIT II
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Page 5
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