Applied Photophysics Chirascan CCD User manual

Applied Photophysics Limited
21 Mole Business Park
Leatherhead
Surrey
KT22 7BA
United Kingdom
Tel: [phone]
Fax: [phone]
Email: [e-mail]
Applied Photophysics, Inc.
100 Cummings Center
Suite 440-C
Beverly
MA 01915
United States
Tel: [phone]
Email: [e-mail]
www.photophysics.com
User Manual
Chirascan CCD Emission Fluorometer

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Chirascan CCD Emission Fluorometer
This document contains important safety information. Read this document, the
Chirascan V100 Spectrometer User Manual and the User Manual provided by
Ocean Optics before attempting to install or use the CCD Emission Fluorometer.
Failure to do so could result in death or serious injury.

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Chirascan CCD Emission Fluorometer
CONTENTS
CONTENTS........................................................................................................................................................3
USE OF THIS DOCUMENT ...............................................................................................................................4
ESSENTIAL SAFETY INFORMATION...............................................................................................................5
CHIRASCAN CCD EMISSION FLUOROMETER INSTALLATION AND OPERATIONAL REQUIREMENTS..6
GLOSSARY........................................................................................................................................................7
1 INTRODUCTION.............................................................................................................................................8
2 INSTALLATION ...............................................................................................................................................9
2.1 Single Cell Peltier Temperature Controller...........................................................................................9
2.2 6-cell Peltier Cell Holder .....................................................................................................................11
2.3 The CCD spectrometer.......................................................................................................................13
3 OPERATION..................................................................................................................................................14
3.1 Typical use..........................................................................................................................................14
3.2 Setting the CCD conditions.................................................................................................................14
3.2.1 CCD only mode .................................................................................................................................. 14
3.2.2 Operation with other signals............................................................................................................. 15
3.2.3 Acquiring a baseline .......................................................................................................................... 17
3.3 Pro-Data Viewer..................................................................................................................................17
3.3.1 Data files............................................................................................................................................ 17
3.3.2 Example: quinine sulfate reference .................................................................................................. 18
3.3.3 Example: anthracene reference ........................................................................................................ 19
3.3.4 Example: -chymotrypsin ................................................................................................................. 20
4 MAINTENANCE AND STORAGE .................................................................................................................22

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Chirascan CCD Emission Fluorometer
USE OF THIS DOCUMENT
This document is intended to inform the operator of Applied Photophysics Chirascan CCD Emission
Fluorometer on its design, installation and operation with the Chirascan V100 Spectrometer and the Single
Cell Peltier Temperature Controller, or the 6-cell Peltier Cell Holder. The CCD Emission Fluorometer is the QE
Pro Scientific-grade type supplied by Ocean Optics Inc., Dunedin, FL, USA, and this document should be used
in conjunction with the User Manuals supplied by Ocean Optics and those applicable to the spectrometer and
the Single Cell Peltier Temperature Controller, or the 6-cell Peltier Cell Holder. It is assumed that the user of
this document is familiar with the operation of the Chirascan spectrometer, and with Applied Photophysics Pro-
Data software. In particular it is assumed that the user is familiar with the hazards associated with the operation
of the spectrometer and the Single Cell Peltier Temperature Controller, or the 6-cell Peltier Cell Holder, and
has read the hazard and other safety information contained in their User Manuals and those supplied by Ocean
Optics.
The information in this document is subject to change without notice and should not be construed as a
commitment by Applied Photophysics, who accept no responsibility for errors that may appear herein. This
document is believed to be complete and accurate at the time of publication, and in no event shall Applied
Photophysics be held responsible for incidental or consequential damages with or arising from the use of this
document.
COPYRIGHT 2018 APPLIED PHOTOPHYSICS LIMITED. ALL RIGHTS RESERVED. THIS DOCUMENT
OR PARTS THEREOF SHALL NOT BE REPRODUCED IN ANY FORM WITHOUT THE WRITTEN
PERMISSION OF THE PUBLISHER.
THE SOFTWARE PROVIDED WITH THE CHIRASCAN AND ITS ACCESSORIES (PRO-DATA CHIRASCAN,
PRO-DATA VIEWER, ETC.) IS THE PROPERTY OF APPLIED PHOTOPHYSICS LIMITED AND IS
SUPPLIED UNDER LICENSE. APPLIED PHOTOPHYSICS IS WILLING TO LICENSE THE SOFTWARE
ONLY UPON THE CONDITION THAT THE LICENSEE ACCEPTS ALL THE TERMS CONTAINED IN THE
LICENSE AGREEMENT. THESE INCLUDE THAT THE LICENSEE MAY NOT SELL, RENT, LOAN OR
OTHERWISE ENCUMBER OR TRANSFER LICENSED SOFTWARE IN WHOLE OR IN PART TO ANY
THIRD PARTY. FOR A FULL COPY OF THE LICENSE PLEASE CONTACT APPLIED PHOTOPHYSICS OR
SEE THE SOFTWARE INSTALLATION DISK.
Chirascan is a trademark of Applied Photophysics Limited.
All other trademarks or registered trademarks are the sole property of their respective owners

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Chirascan CCD Emission Fluorometer
ESSENTIAL SAFETY INFORMATION
MAKE SURE THAT YOU HAVE READ AND UNDERSTOOD ALL THE SAFETY INFORMATION CONTAINED
IN THE MAIN CHIRASCAN USER MANUAL AND THE USER MANUAL PROVIDED BY OCEAN OPTICS
BEFORE ATTEMPTING TO INSTALL OR OPERATE THE CHIRASCAN CCD EMISSION FLUOROMETER.
IF YOU HAVE ANY QUESTIONS REGARDING THE OPERATION OF THE FLUOROMETER, PLEASE
CONTACT APPLIED PHOTOPHYSICS TECHNICAL SUPPORT AT THE ADDRESS SHOWN ON THE FIRST
PAGE OF THIS DOCUMENT.
OBSERVE ALL SAFETY LABELS AND NEVER ERASE OR REMOVE SAFETY LABELS.
PERFORMANCE OF INSTALLATION, OPERATION OR MAINTENANCE PROCEDURES OTHER THAN
THOSE DESCRIBED IN THIS USER MANUAL MAY RESULT IN A HAZARDOUS SITUATION AND WILL
VOID THE MANUFACTURERS WARRANTY.

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Chirascan CCD Emission Fluorometer
CHIRASCAN CCD EMISSION FLUOROMETER INSTALLATION AND OPERATIONAL
REQUIREMENTS
Environmental and electrical requirements
The Chirascan CCD Emission Fluorometer has no environmental requirements additional to those of the
Chirascan spectrometer, but will require a separate mains voltage supply.
Bench space
The Chirascan CCD Emission Fluorometer requires an additional bench space of about 150 mm depth and
200 mm width to the right of the Chirascan sample handling unit, but is low enough to fit below the Chirascan
detector.
Nitrogen purge gas
The Chirascan CCD Emission Fluorometer has no purge gas requirements additional to those of the Chirascan
spectrometer.
Circulating water
The Chirascan CCD Emission Fluorometer has no circulating water requirements additional to those of the
Chirascan spectrometer.
Servicing
Servicing of the Chirascan CCD Emission Fluorometer should only be undertaken by qualified personnel. If
you are in any doubt at all please contact the Applied Photophysics’Technical Support at the address given
on the front of this User Manual.

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Chirascan CCD Emission Fluorometer
GLOSSARY
The following abbreviations may be found in this User Manual
APL Applied Photophysics Limited
CCD charge-coupled device
CD circular dichroism
IR infrared
SHU sample handling unit
UV ultraviolet
HYPERLINKS
This document contains hyperlinks between references (for example the Contents tables, or references to
Sections or Figures in the text), and sources. To follow a link, place the cursor over the reference and use
CTRL+click. Hyperlinks in the text are indicated by underlined blue font.

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Chirascan CCD Emission Fluorometer
1 INTRODUCTION
The Chirascan CCD Emission Fluorometer is designed to be used with a Chirascan V100 Spectrometer and
the Single Cell Peltier Temperature Controller or the 6-cell Peltier Cell Holder. The detector is shown in Figure
1.1, with fitting for connecting to a Single Cell Peltier Temperature Controller.
Figure 1.1: the Chirascan CCD Emission Fluorometer
Fluorescence occurs when electromagnetic radiation absorbed by a sample is re-emitted at a longer
wavelength. With the Emission Fluorometer the emitted light is collected from the fluorescence window of the
Single Cell Peltier Temperature Controller or the 6-cell Peltier Cell Holder, and focused onto an optical fiber
light guide which takes it to a CCD spectrometer. The CCD spectrometer unit contains a grating that disperses
the incoming light and directs it onto a CCD detector with a 2-D arrangement of pixels (1044 horizontal x 64
vertical) that is responsive from 200 nm to 1000 nm. A full fluorescence spectrum can be acquired in just a few
seconds.
The CCD spectrometer is the QE Pro Scientific-grade mini-spectrometer type supplied by Ocean Optics Inc.,
Dunedin, FL, USA, and is supplied with a manual provided by Ocean Optics.

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Chirascan CCD Emission Fluorometer
2 INSTALLATION
2.1 Single Cell Peltier Temperature Controller
The fittings required for connecting the Emission Fluorometer to the Single Cell Peltier Temperature Controller
are shown in Figure 2.1.
Figure 2.1: the fittings required for connection to the single cell accessory,
left to right: dovetail insert, grub screw and nut, lens holder, light guide ferrule
Figure 2.2: the dovetail insert mounted on the Single Cell Peltier Controller
The dovetail insert mounts onto the Single Cell Peltier Controller fluorescence window as in Figure 2.2 (for
clarity, the Single Cell Peltier Controller is shown removed from the Chirascan sample handling unit). To hold
the insert in place, tighten the 16 mm grub screw that screws into the threaded hole in the center using a 1.5
mm hexagon key. Do not over tighten the grub screw, it is enough just to hold the dovetail in place.
16 mm grub screw
Hole for lens holder

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Chirascan CCD Emission Fluorometer
Figure 2.3: light guide collar with the light guide inserted.
Now insert the tip of the light guide through the grommet on the guide collar; the guide tip should protrude
about 3 cm through the grommet. Screw the light guide ferrule onto the tip of the guide (Figure 2.4).
Insert the ferrule into the lens holder, ensuring that it is fully seated home. Hold the lens holder in place by
tightening the two grub screws on the holder using a 1.5 mm hexagon key. Do not over tighten the grub screws;
they are only to hold the light guide in the lens holder.
The light guide collar mounts in the fluorescence port of the Chirascan sample handling unit as in Figure 2.4.
The lens holder inserts into the hole in the dovetail insert, with the 16 mm grub screw locating in the groove in
the lens holder. An M3 nut on the grub screw is used to hold the lens holder in place. For clarity, this
arrangement is shown with the Single Cell Peltier Holder removed from the Chirascan sample handling unit in
Figure 1.1.
Figure 2.4: the light guide mounted on the Chirascan sample handling unit,
Light guide
Light guide ferrule
Light guide collar
Grommet
Light guide
collar
CD detector
Lens holder