Applied Photophysics RX2000 User manual
User Manual
RX2000
Rapid Mixing Stopped Flow Unit
April 2013
Document 4400Q902 version 1.02
Document 4400Q902 version 1.02 Page 2 of 18
RX2000 User Manual
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RX2000 User Manual
This document contains important safety information. Read this document before
attempting to install or use the RX2000. Failure to do so could result in injury to the
user.
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RX2000 User Manual
USE OF THIS DOCUMENT
This document is intended to inform the operator of Applied Photophysics’ RX2000 Rapid-Mixing Stopped-Flow
Unit on its design, installation and operation. 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 2013 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.
RX2000™is a trademark of Applied Photophysics Ltd.
Norton™ is a trademark of Symantec Corporation or its affiliates in the United States and other countries.
McAfee® is a registered trademark of McAfee, Inc. in the United States and other countries.
Sophos® is a registered trademark of Sophos Plc and Sophos Group.
Kloehn® is a registered trademark of Kloehn Inc.
All other trademarks or registered trademarks are the sole property of their respective owners.
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RX2000 User Manual
HAZARD AND OTHER INDICATORS
HAZARD INDICATORS USED IN THIS DOCUMENT
The sign to the left is used to indicate a hazardous situation, which, if not avoided, could result in
minor or moderate injury.
OTHER INFORMATORY INDICATORS USED IN THIS DOCUMENT
The sign to the left is used to indicate a situation which, if not avoided, could result in damage to
the instrument.
HAZARD INDICATORS USED ON THE RX2000
Note that this hazard indicator may be either coloured as below or as black and white.
The sign to the left is a general hazard indicator, indicating the presence of a hazard that is either
described by text accompanying the sign or in this User Manual.
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RX2000 User Manual
ESSENTIAL SAFETY INFORMATION
MAKE SURE THAT YOU HAVE READ AND UNDERSTAND ALL THE SAFETY INFORMATION CONTAINED
IN THIS DOCUMENT BEFORE ATTEMPTING TO OPERATE THE RX2000. IF YOU HAVE ANY QUESTIONS
REGARDING THE OPERATION OF THE RX2000, PLEASE CONTACT APL TECHNICAL SUPPORT SECTION
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.
The RX2000 pneumatic accessory uses compressed gas to drive the drive syringes and could
trap hands or clothing, causing injury to the user. Keep hands and clothing clear when
performing a drive using the accessory.
Corrosive chemical and organic solvents can cause damage to the RX2000. Do not allow
corrosive fluids to come into contact with any part of the spectrometer. Do not clean the
spectrometer with organic solvents. Use only a soft cloth and water or a mild detergent solution.
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RX2000 User Manual
RX2000 INSTALLATION AND OPERATIONAL REQUIREMENTS
Environmental requirements
The RX2000 is best installed in a safe position, in a clean, air-conditioned laboratory environment.
Operating conditions
Temperature: ±2oC of a fixed temperature in the range 18oC to 26oC.
Humidity 20% to 80% non-condensing
Storage conditions
Temperature -20°C to +50°C:
Humidity: 5% to 80% non-condensing
Bench space
The RX2000 requires a bench space of
Length x depth x height: 0.7 x 0.3 x 0.2 metres
alongside the spectrometer with which it is used.
A sturdy, stable, vibration-free work surface is recommended.
Fluid circulator unit
The RX2000 can be used with or without temperature control. Temperature control is provided by a standard
laboratory temperature controlled fluid circulator.
Servicing
Servicing of the RX2000 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
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RX2000 User Manual
CONTENTS
USE OF THIS DOCUMENT.......................................................................................................................................4
HAZARD AND OTHER INDICATORS.......................................................................................................................5
ESSENTIAL SAFETY INFORMATION......................................................................................................................6
RX2000 INSTALLATION AND OPERATIONAL REQUIREMENTS..........................................................................7
CONTENTS ...............................................................................................................................................................8
FIGURES...................................................................................................................................................................8
1 INTRODUCTION ....................................................................................................................................................9
2 HARDWARE.........................................................................................................................................................10
2.1 Basic layout................................................................................................................................................10
2.2 The drive syringes......................................................................................................................................11
2.3 Reagent control valves...............................................................................................................................11
2.4 Waste control valve....................................................................................................................................12
2.5 The observation cell...................................................................................................................................13
2.6 Accessories................................................................................................................................................13
2.7 RX2000 specifications................................................................................................................................13
3 OPERATION.........................................................................................................................................................14
3.1 Instrument set-up .......................................................................................................................................14
3.1.1 Positioning the mixing unit and the DuoCell ............................................................................................. 14
3.1.2 Temperature control.................................................................................................................................. 14
3.1.3 Trigger signal.............................................................................................................................................. 14
3.2 Operation using the manual drive..............................................................................................................14
3.3 Operation using the pneumatic drive accessory........................................................................................15
3.4 Operation using the anaerobic accessory..................................................................................................16
3.4.1 Mounting the anaerobic accessory............................................................................................................ 16
3.4.2 Anaerobic operation .................................................................................................................................. 16
3.5 Operation using ratio mixing ......................................................................................................................17
3.6 Changing the drive syringe. .......................................................................................................................17
3.7 Selected test reactions...............................................................................................................................17
3.7.1 Formation of Iron (III) Thiocyante.............................................................................................................. 17
3.7.2 Fluorescence test reaction......................................................................................................................... 18
FIGURES
Figure 1.1: the RX2000.............................................................................................................................................9
Figure 2.1: schematic of the RX2000 .....................................................................................................................10
Figure 2.2: the valve positions in the Load (left) and Drive (right) positions...........................................................11
Figure 2.3: flow line closed (left) and open to waste reservoir (right).....................................................................12
Figure 2.4: flow line in its normal drive position (left) and normal empty position (right)........................................12
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RX2000 User Manual
1 INTRODUCTION
Figure 1.1: the RX2000
The RX2000, shown in Figure 1.1, is a high performance stopped-flow unit that enables your existing UV-visible
spectrometer, fluorimeter or circular dichroism (CD) spectrometer to be used to follow reactions that are at least
a thousand times faster than those accessible manually. With a dead-time of only 8 ms, first order reactions with
rates in excess of 200 s-1 can be followed.
The RX2000 is easily the most accomplished device of its type, making use of the latest advances in stopped-
flow design, combined with the simplicity, robustness and reliability of high quality instrument engineering. In one
action the RX2000 mixes two reactants, fills a sample cell, stops the flow and simultaneously provides an output
trigger. Its dual pathlength micro-volume cell employs the latest silica fusion technology to give the highest
possible optical efficiency and specification. All crucial sample flow circuit surfaces are biocompatible and
chemically inert. In addition, carefully selected materials are used throughout to protect the instrument from
aggressive chemicals. The entire flow circuit and the drive syringes themselves are located within a
thermostatable bath for accurate sample temperature control.
The stopped-flow technique for the rapid mixing of chemical reagents has gained widespread acceptance as
being a most useful and versatile method for obtaining reaction rate information. After the two reagents are
mixed, the course of their reaction together can be followed in conjunction with single beam UV-visible
absorption or fluorescence spectrometry as appropriate, using the optical absorption or emission properties of
either the initial reagents, or a product or a reaction intermediate. Changes in light scattering within the reaction
cell provide another measurement method. The technique finds applications in chemistry (redox reactions, metal-
ligand complexes, solvation, dye binding etc.), biochemistry (enzyme kinetics, electron and proton reactions,
binding of poisons to biologically important molecules, calcium and sodium ion interactions, protein binding,
enzyme conformation and denaturation studies) and biophysics (cell membrane studies, osmotic swelling,
oxygen uptake and release by red blood cells).
This User Manual gives comprehensive information about how to configure and operate the RX2000 Rapid
Mixing Stopped-Flow Unit.
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RX2000 User Manual
2 HARDWARE
This Chapter describes the RX2000 hardware, covering the basic layout and main components of the accessory.
2.1 Basic layout
Figure 2.1: schematic of the RX2000
A schematic of the RX2000 is shown in Figure 2.1. The unit is designed to be compact and easy to use, allowing
stopped-flow measurements to be obtained with UV-visible spectrometers, fluorimeters and CD polarimeters with
the minimum amount of setting-up. It comprises a base in which the electronics, battery and LCD temperature
display panel are located. Attached to the top of the base is a reagent handling system with thermostat bath and
this is connected to a specialised cuvette via an umbilical. The sealed thermostat bath incorporates a window
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