User Manual Chirascan CS/LD
Contents
1. RHOOCTC pump supplied by Fluid Metering Inc Syosset NY USA The spectra have been corrected for background spectrum with the pump turned off and a 3 point smooth has been applied to even out the effect of the pulsing pump motion Document 4207Q116C03 01 Page 12 of 122. SOFTWARE IN WHOLE OR IN PART TO ANY THIRD PARTY FOR A FULL COPY OF THE LICENCE PLEASE CONTACT APL OR SEE THE SOFTWARE INSTALLATION DISC Chirascan and Chirascan plus are trademarks of Applied Photophysics Ltd Microsoft and Windows are registered trademarks of Microsoft Corporation in the United States and other countries All other trademarks or registered trademarks are the sole property of their respective owners Document 4207Q116C03 01 Page 4 of 12 Applied Ultrasensitive Spectroscopy for the Life Sciences CS LD Accessory User Manual HAZARD AND OTHER INDICATORS HAZARD INDICATORS USED IN THIS DOCUMENT AWARNING The sign to the left is used to indicate a hazardous situation which if not avoided could result in death or serious injury OTHER INFORMATORY INDICATORS USED IN THIS DOCUMENT leya 4 The sign to the left is used to indicate a situation which if not avoided could result in damage to the instrument Document 4207Q116C03 01 Page 5 of 12 Applied Ultrasensitive Spectroscopy for the Life Sciences CS LD Accessory User Manual ESSENTIAL SAFETY INFORMATION MAKE SURE THAT YOU HAVE READ AND UNDERSTAND ALL THE SAFETY INFORMATION CONTAINED IN THIS DOCUMENT BEFORE ATTEMPTING TO INSTALL OR OPERATE THE LINEAR DICHROISM ACCESSORY IF YOU HAVE ANY QUESTIONS REGARDING THE OPERATION OF THE ACCESSORY PLEASE CONTACT APL TECHNICAL SUPPORT SECTION AT THE ADDRESS SHOWN ON THE FIRST PAGE OF THIS DOCUMENT OBSERVE ALL
3. AppliedPhotophysics Ultrasensitive Spectroscopy for the Life Sciences User Manual Chirascan CS LD Chirascan Series Linear Dichroism Accessory February 2012 Document 4207Q116C03 01 o Photophysics C2272 j Applied Photophysics Ltd 21 Mole Business Park Leatherhead Surrey KT22 7BA UK Tel UK 44 1372 386 537 Tel USA 1 800 543 4130 Fax 44 1372 386 477 Applied Photophysics was established in 1971 by The Royal Institution of Great Britain WWW PD hoto D hys ics com Applied hotophysics Ultrasensitive Spectroscopy for the Life Sciences EPEE PE E vo vaen ee EEES CS LD Accessory User Manual This page is intentionally left blank Document 4207Q116C03 01 Page 2 of 12 Applied Ultrasensitive Spectroscopy for the Life Sciences PE ARA ees CS LD Accessory User Manual This document contains important safety information Read this document and the Chirascan series User Manual before attempting to install or use the CS LD accessory Failure to do so could result in death or serious injury Document 4207Q116C03 01 Page 3 of 12 Applied Ultrasensitive Spectroscopy for the Life Sciences E P ea ee CS LD Accessory User Manual USE OF THIS DOCUMENT This document is intended to inform the operator of Applied Photophysics CS LD linear dichroism accessory on its design installation and operation The CS LD is used with either the Chirascan or Chirascan plus spectrometer and this document
4. Document 4207Q116C03 01 Page 10 of 12 Applied Ultrasensitive Spectroscopy for the Life Sciences saan ee ieee Bosco esas CS LD Accessory User Manual 2 INSTALLATION AND OPERATION 2 1 Hardware installation WARNING The Linear Dichroism photomultiplier tube PMT and Large Area Photoediode LAAPD detector operate at high voltages can produce a shock leading to serious injury or death Do not connect or disconnect the detector from the spectrometer unless the spectrometer is powered off The detectors currently April 2011 onwards supplied with the Chirascan or Chirascan plus spectrometers can also be used in CD mode and their installation is described in the main Chirascan User Manual 2 2 Configuration and calibration The detector needs to configured and calibrated for LD measurements If the LD option was purchased with the spectrometer this will have been performed by APL as part of the production process If the LD option is purchased as an upgrade then the configuration and calibration will need to be performed by an APL qualified engineer leyg A Incorrect configuration of the detector can cause damage to the instrument or give incorrect results and LD calibration of the detector requires a special rig Configuration and calibration should only be performed by a qualified APL engineer Note that the calibration factors are specific to a particular detector Users upgrading from a single mode CD detector should not use th
5. 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 WARNING The Chirascan spectrometer is powered by the mains electricity supply which can produce a shock leading to serious injury or death Do not connect or disconnect the instrument from the mains supply unless the supply is powered off at source Ensure all communications and electrical connections are made before powering on the spectrometer Exercise care during operation and do not operate units with their covers removed Operate the spectrometer using only the cables provided Never operate a spectrometer with damaged cables WARNING The Linear Dichroism photomultiplier tube PMT and Large Area Photodiode LAAPD detectors operate at high voltages can produce a shock leading to serious injury or death Do not connect or disconnect the detector from the spectrometer unless the spectrometer is powered off Document 4207Q116C03 01 Page 6 of 12 Applied Ultrasensitive Spectroscopy for the Life Sciences E P ea ee CS LD Accessory User Manual CS LD INSTALLATION AND OPERATIONAL REQUIREMENTS Environmental and electrical requirements The CS LD has no environmental requirements additional to those of the Chirascan or Chirascan plus spectrometer Operation of the CS LD is controlled by t
6. e single mode CD detector and the CD LD detectors interchangeably 2 3 Operation From the drop down list on the Signal panel on the Chirascan Pro Data control page select Linear Dichroism Figure 2 1 From then on operation of the instrument is similar to that when making CD measurements for details of which see the main Chirascan series User Manual Signal Linear Dichroism gt del A he Options i Absorbance Fluorescence Bazeline Baseline E Bea Recall Figure 2 1 the Signal panel Document 4207Q116C03 01 Page 11 of 12 Applied hotop Ultrasensitive Spectroscopy for the Life Sciences ee ree CS LD Accessory User Manual Typically when making LD measurements a baseline is taken with no sample present and LD spectra are then measured with the sample present but no flow and then at selected flow rates Background correction is made by subtracting the spectrum measured without flow from those measured with flow co Co KI 0 015 mm m 0 005 Linear Dichroism delta A 220 230 240 250 260 270 280 290 300 VVavelength nm Figure 2 2 LD spectra for calf thymus DNA at three different flow rates Results for calf thymus DNA at a concentration of 1 mg ml in phosphate buffered saline are shown in Figure 2 2 at three different pump speeds 30 ml min orange 15 ml min red and 7 5 ml min cyan A 0 5 mm pathlength x 4 mm width flow cell of the type shown in Figure 1 1 was used with an
7. he Chirascan electronics and there are no additional electrical requirements Bench space The CS LD has no bench space requirements additional to those of the Chirascan spectrometer although some users may choose to use an external device to generate flow in the LD which may add to the bench space requirement Nitrogen purge gas The CS LD has no purge gas requirements additional to those of the Chirascan spectrometer Servicing servicing of the CS LD should only be undertaken by qualified personnel If you are in any doubt at all please contact the Applied Photophysics Technical Support Department at the address given on the front of this User Manual Document 4207Q116C03 01 Page 7 of 12 Applied Ultrasensitive Spectroscopy for the Life Sciences E A P AA E CS LD Accessory User Manual GLOSSARY The following abbreviations may be found in this User Manual APL Applied Photophysics Ltd CD Circular dichroism LD Linear dichroism PMT Photomultiplier tube 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 Document 4207Q116C03 01 Page 8 of 12 Applied Ultrasensitive Spectroscopy for the Life Sciences CS LD Accessory User Manual CONTENTS U EOF TAD DOCUMEN caes E E E teen eee e
8. ne eee eee eee ee ee ee eee ee 4 HAZARD AND OTHER INDICA TOR marno nennen E T ETE ERE 5 ESSENTIAL SAFETY INFORMATION cicsccinsierscsiccievecevecenceesceacech neeencceteiscuecs nen cnoetisaceancd senecose eseunceredceesbsasencedioneseuebendnes 6 CS LD INSTALLATION AND OPERATIONAL REQUIREMENTS 0 cccccccseceeeeeeeeeeeeeeeeseeeeeeeeseeaeeeeeseseaaeeeeesseaaes 7 GEOS ANY ora E E E EE E SE E E E 8 CONTENT arse orate cence cece AEA E ITE E AEE O E geo 9 PIO URE are E R E A E 9 TINTRODUG MON oireeni ne net ro ee E E A E EE E 10 2 INSTALLATION AND OPERATION siccccccccceiccnedsncntionsendssneiendetjocasnananedsnsintensendannesandatewsenspacedendehiewaensanesdciedeeeusencnenndecs 11 2 1 Hardware installation cccccccccccssssececeeeeeeeeeeeeeeeeeeeeeeeeeeeesseeeeeseaeeeeseaueeessaeeeesseeeeeseaseeeseaeeeessegeeesseneeeeseess 11 2 2 Configuration and calibration cccccccsseeeeececceeceeeeeeeeceeeeeeeeeeeeeeeeeeeesseeeeeeeeeeeeesseaeseeeeeeeessaaeseeeeeeeessaagasess 11 2 ODO ON co cee aea ea DEE A a N e EEE EE EEE O EEE E T N EE EEE E 11 FIGURES Figure 1 1 flow cell of the type commonly used with the CS LD ec cceccccccccsseeeeeeeeaeeeeeeeeseeeeeeeeeeaaseeeessaaeeeees 10 Figure 2 1 the Signal panel n a1ennoannnnn1nnnnnnnnrnrnrernnnnrnrnrernnanrnrnnrrnnnnrnnnrennnansnrnteonannrnrnrennnansnanneennnnrnnnrennnnnrnnnn eee nn ee 11 Figure 2 2 LD spectra for calf thymus DNA at three different flow rates 0 0 0 ccccceecceec
9. seeeeeeseeeeeeeeeeeeeeeseeessaeeeeens 12 Page 9 of 12 Document 4207Q116C03 01 Applied Ultrasensitive Spectroscopy for the Life Sciences CS LD Accessory User Manual 1 INTRODUCTION Linear dichroism is the difference in the absorbance of parallel and perpendicular linearly polarized light Equation 1 1 It is usually produced by preferential orientation of macromolecules for example by stretching along the streamlines in a flow field AA Aj A 1 1 where A is the absorbance of the linearly polarised light and the subscripts I and L indicate parallel and perpendicular respectively On older Chirascan and Chirascan plus spectrometers linear dichroism measurements required a different detector from that used for circular dichroism measurements On current instruments April 2011 onwards the same detector may be used for both although it needs to be configured and calibrated for each mode This should be done by a qualified APL engineer Figure 1 1 flow cell of the type commonly used with the CS LD Sampling handling and orientation are left to the discretion of the user Commonly a flow through cell such as that shown in Figure 1 1 available from Hellma UK Ltd is used The use of a concentric cylinder Couette type flow cell is also well established For more information on linear dichroism and its measurement see B Norden A Rodger and T Dafforn Linear Dichroism and Circular Dichroism RSC Publishing 2010
10. should be used in conjunction with the User Manual applicable those instruments It is assumed that the user of this document is familiar with the operation of the Chirascan or Chirascan plus 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 has read the safety information contained in its User Manual 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 2012 APPLIED PHOTOPHYSICS LTD 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 LTD APL AND IS SUPPLIED UNDER LICENCE APL IS WILLING TO LICENSE THE SOFTWARE ONLY UPON THE CONDITION THAT THE LICENSEE ACCEPTS ALL THE TERMS CONTAINED IN THE LICENCE AGREEMENT THESE INCLUDE THAT THE LICENSEE MAY NOT SELL RENT LOAN OR OTHERWISE ENCUMBER OR TRANSFER LICENSED
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