Jenway 6285 User Manual

Fluorimeters

Models 6270, 6280 & 6285

Operating Manual

628 020/REV A/04- 06

Safety

This is important information; please read carefully before installing or using this
instrument.

1. The 6

2. The fluorimeter is a sensitive electronic and optical instrument designed for use in a

3. In addition to observing the instructions detailed in the Operating Manual and Service

4. Voltage levels hazardous to life are present in this instrument, for personal safety only

5. This instrument is designed for minimal maintenance, which must be carried out carefully

6. Other than for those items defined in the maintenance procedures herein there are no

7. Reference should always be made to the Health and Safety Data for any chemicals or

8. It is important that good laboratory practice is observed when handling samples,

9. If it is suspected that safety protection has been impaired in any way, the fluorimeter must

2 Series Fluorimeters are designed for operation by trained personnel that are

aware of the principles and applications involved. For further help and advice please
contact your local distributor, e-mail [e-mail]

or visit www.jenway.com

laboratory environment. Careful adherence to the installation instructions must be

observed. If in doubt contact a relevant and competent authority for advice before

proceeding.

Manual for this instrument all installation, operating and service personnel must be aware

of, and employ, a safe system of work.

trained engineers aware of the risk and avoidance of electric shock should remove

protective covers from the instrument.

following the procedures detailed in this manual. All safety instructions in these
procedures, as well as those defined locally for the area or environment where the work

is being carried out must be observed.

user serviceable items in this instrument. Removal of covers and attempted adjustment

or service by unqualified personnel will invalidate any warranty and incur additional
charges for repair.

reagents used. All available information, advice and warnings on the handling, storage,
use and disposal of such must be carefully observed. When not available this data must
be requested from the supplier before proceeding in any way.

chemicals, reagents and ancillary equipment in order to carry out measurement and

analysis with this instrument. Suitable safety and personal protective equipment must

be used at all times.

be made inoperative and secured against any intended operation. The fault condition
must be reported to the appropriate servicing authority. In all such reports the model

number and serial number of the fluorimeter must be quoted.

Contents:

Section 1 Introduction

Instrument description
Good practice guidelines

Section 2 Getting Started

Unpacking
Installation

Section 3 Fluorescence Theory
Section 4 Instrument Menu Options
Section 5 Instrument Settings

Instrument Lock

Date, Time and Language
Security

Section 6 Setting up a method
Section 7 Using a stored method
Section 8 Calibration
Section 9 Performing Measurements

Section 10 Timed Readings
Section 11 Saving Results
Section 12 Printing Results
Section 13 Maintenance & Troubleshooting

General and routine maintenance
Error codes and troubleshooting

Section 14 Accessories and Spares

Optional accessories

Section 15 Specification & Data

Technical specification
RS232 serial interface

EC Declaration of Conformity

Operator ID

Glossary of Terms

Section 1

Introduction

1.1 Instrument Description

Three models are available to cover a wide range of applications. The model 6280
covers the most sensitive determinations with emission wavelengths up to 650nm.
Where higher emission wavelengths will be utilised the model 6285 with its red­enhanced detector is applicable. For less sensitive applications with a broader
wavelength range the model 6270 will meet the necessary requirements.

All three models offer intuitive operation with a user interface based on logical menus
that can be navigated from the simple keypad. Up to 20 methods can be created
without restriction and be saved for future use, they can also be locked against
accidental change by password entry, ensuring data integrity.

The permanent time and date tag on every stored reading further enhances Good
Laboratory Practice, while calibration reminders and operator identity can also be
entered to support conformance and traceability of operation.

With press-to-read operation and Total energy transfer (TET) technology the output
of the high-energy xenon lamp is maximised and its expected life extended so that it
should never need replacing in normal use.

The high quality optics is complemented by the Intelligent Filter Modules (IFM) that
can be identified by the system, this enables error messages to be generated and
displayed should the wrong filters be fitted or their positions be incorrect.

All models are powered from an external, universal mains adapter suitable for use
from 90Vac to 264Vac, while the 12V dc input enables use of the fluorimeters in
vehicles or from suitable battery packs.

Where temperature controlled fluorescence studies are necessary an optional
electrically heated sample holder is easily fitted while for continuous flow analysis an
external sipper pump and a wide range of flow through cuvettes are also available.

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1.2 Good Practice Guidelines

1. The selection of the optimum excitation and emission wavelengths (filters) is

critical to achieving the best performance from the analysis.

2. All fluorimeters should be sited in a clean, dry, dust free environment. When in
use ambient temperature and light levels should remain as constant as possible.

3. Adherance to Standard Operating Procedures (SOP) and Good Laboratory
Practice (GLP) should be maintained, with regular calibration checks and a suitable
Quality Control (QC) programme.

4. The correct selection of cuvettes is imperative for accurate and reproducible
results:

a) Ensure all cuvettes used are compatible with fluorimetric measurements

where the emission beam is at 90° to the excitation source. Typical examples
have 4 clear sides.

b) Ensure any native fluorescence from the cuvette material is minimal at the

analysis wavelengths.
c) Plastic cuvettes should be used once only.
d) Glass and quartz cuvettes should be thoroughly cleaned after use. Discard

when scratches or marks are evident in their optical surfaces.
e) Ensure any cleaning agents used do not fluoresce at the analytical

wavelengths and are thoroughly rinsed away before drying.
f) Ensure the cuvettes used are compatible with the constituents of both the

samples and standards they are to hold. Plastic cuvettes are not compatible

with some organic solvents.
g) Cuvettes must be handled with care; by the top and non-optical surfaces only.

Any finger marks must be removed by using a suitable cleaning process.
h) Flow through cuvettes must be selected with additional consideration for the

sample type, aspirated volume, pumping system and rinse cycle, as well as

the waste handling to be used.

5. The high sensitivity of fluorimetric analysis means that all glassware used in the
preparation of samples and standards must be totally free from contamination.

6. Chemicals and reagents used in sample and standard preparation should be of
the highest grade of purity (AR Grade) and all should be checked for excessive
background fluorescence at the analytical wavelengths.

7. Samples and working standards should not be stored due to the effects of
evaporation, as well as chemical and photo-degradation. Only prepare samples and
working standards when they are required for analysis.

8. Fluorescence is inversely proportional to temperature. Ensure that all samples
and standards have equilibrated to ambient temperature before analysis. If in doubt,
use a temperature controlled cuvette holder.

9. Refrigerated or cold samples will form micro-bubbles on the cuvette wall as they
warm up. These are a common cause of drift in readings. Ensure all samples and
standards have equilibrated to ambient temperature before analysis.

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10. Check the linear range for each method and, where necessary, use a multi-point
calibration or calibration curve.

11. Be aware of the effects of quenching and, where necessary, use sample dilution
or extraction methods to eliminate this.

12. Monitor the blank during and between batches to identify any increase due to
contamination.

13. Sources of contamination to be considered include cleaning agents, micro­organisms, particles in suspension, stop-cock grease, filter paper residues and
plasticisers leached from containers, caps or sealing materials.

14. Many fluorescent assays are pH dependent. Ensure the pH of all samples and
standards is within specified limits before carrying out the analysis.

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Section 2

Getting Started

2.1 Unpacking

Remove the universal 12V power supply (with UK, US and EU leads) and the pack of
100 disposable cuvettes from the packaging.

Remove the fluorimeter from the carton by lifting it in the centre between the two
support cheeks; do not lift it by the support cheeks.

Place all items on a clean workbench then remove the support cheeks and the
polythene bag from the fluorimeter.

Any shortages or damage must be reported to your local distributor or the
manufacturer as soon as possible.

Keep all packing materials in case the unit has to be re-shipped at a later date.
It is important that when re-packing the instrument it is first sealed in a strong, clean
polythene bag to protect it from the dust and particles that are present in all packing
materials.

2.2 Installation

2.21 Location

In ideal circumstances the installation environment will be clean, dry and dust free
with the instrument protected from extreme variations in ambient lighting and
temperature change. For field use it is recommended that the instrument is used in
the optional storage case for additional protection.
Where conditions are less than ideal, maintenance and cleaning must be carried out
regularly and additional protection offered where possible. The optional dust cover
should always be fitted when the unit is not being used or is stored for short periods.

2.22 Supply Voltage

The fluorimeter is powered by a low voltage dc power supply that operates from a 90­264Vac mains supply. The universal power supply is supplied with 3 mains leads for
UK, EU and US sockets. The correct lead for your supply should be selected.

2.23 Mains Connections

The leads supplied have a moulded on plug. However, if this is removed for any
reason the wires in the mains lead are colour coded to conform to the internationally
recognised standard such that:

UK CONNECTIONS
BROWN LIVE BLACK LIVE
BLUE NEUTRAL WHITE NEUTRAL
GREEN/YELLOW EARTH GREEN EARTH

Safety
When disposing of any removed plug the connectors must be removed or
made incapable of insertion into a mains socket.

US CONNECTIONS

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2.24 Keypad Functions

1

425673135

246

1. UP ARROW key - used to navigate through menus, to increase values

and for paging up in stored results.

2. DOWN ARROW key - used to navigate through menus, to decrease values

and for paging down in stored results.

3. LEFT ARROW key - used to navigate through menus and to highlight the

selected digit when setting values.

4. RIGHT ARROW key - used to navigate through menus and to highlight the

selected digit when setting values.

5. ENTER key - used to accept the highlighted menu option.

6. CAL key - initiates a calibration sequence from within the

measurement mode.

7. PRINT key - initiates a print from the measurement display or stored

results. Sends data to RS232 serial port.

2.25 Instrument Display

1. Method Name - this will appear on all measurement screens, with the exception of

Raw Fluorescence.

2. Results display - provides direct readout of standards and sample results.

3. Units of measure - shows selected measurement units: ppm, RFU, U/ml, mU/l,

U/l, µM/l, mM/l, M/l, µg/ml, mg/ml, µg/dl, mg/dl, g/dl, ng/l, µg/l, mg/l, g/l, none, %, ppb

4. Status message - shows the current instrument status, such as reading … or

printing … and provides a reminder when calibration is due.

5. Gain - shows the current photomultiplier tube gain setting (0-100%). Gain should

be optimised either manually or by using auto set gain for each method.

Note: This option is not available on the Model 6270.

6. Menu Options - EXIT - allows the user to return to the main menu,
READ - press read to read the sample or standard, SAVE - saves the currently

displayed result to instrument memory

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2.26 Rear Panel Layout

1. Output Socket – 9 way output socket for RS232

2. Switch – Power On/Off switch

3. Connection Sockets – 4 x pin sockets for heated cell block controller.

4. Power In Socket – Connection socket for 12V DC universal mains adapter

2.27 Sample Chamber Filter and Cuvette Positioning

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2.28 Power on and Self-Tests

Connect the mains supply cable to the rear panel mains input socket and plug the
other end into a suitable mains supply socket.

Lift the sample chamber lid on the instrument and ensure that there is no sample or
other item present in the sample holder, close the lid.

Switch on the supply socket, then the instrument, using the Power switch located on
the rear panel.

The instrument will then perform a power on self-test protocol. The following screen
will be shown for approx. 3 seconds:

Please ensure
sample chamber
door remains
closed during
power on tests

Followed by:

On successful completion of these tests the Main Menu screen will be displayed.
For optimum performance a 10-minute warm-up period is required if the ambient
temperature is below 10°C. The unit must be re-calibrated and sample measurement
repeated if this calibration check shows excessive drift.

Should a problem occur during the self-tests an information box or error support
message will be displayed. For assistance please refer to the troubleshooting section
in this manual.

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Example Menu Operation

A common operating system is used throughout this and similar Jenway products, a
brief overview of navigating through the menu system with the cursor keys follows;

Use the up and down arrow keys to highlight a menu option, press the enter key to confirm.

Use the right and left arrow keys to select a digit for adjustment with the up and down arrow

Left arrow

Up arrow key

Right arrow

Down arrow

Enter key

Left arrow

Up arrow key

Right arrow

Down arrow

keys, press the enter key to confirm the setting when ALL digits have been correctly set.
Use the up or down arrow keys to browse through pre-set options, when the correct selection

is displayed press the enter key to confirm your choice.

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Enter key

Left arrow

Up arrow key

Right arrow

Down arrow

Enter key

Section 3

Fluorescence Theory

The interaction between electromagnetic radiation and matter provides a useful,
qualitative and quantitative analytical tool, known as spectroscopy. The region of the
electromagnetic spectrum, to which matter under investigation is subjected to,
defines the type of transitions that occur within the molecules.
Fluorimetry uses radiation from the UV-Vis region of the electromagnetic spectrum to
study transitions between electronic levels in a molecule or atom. The absorption of
energy from light radiation (photons) by a molecule or atom, promotes electrons from
a low energy ground state to a higher energy excited level. This is known as
excitation and the amount of energy transferred to the molecule or atom will depend
on two main factors. The composition of the matter under investigation and the
energy and wavelength of the radiation, have a significant effect on the transitions of
electrons.
The molecule or atom converts the excitation energy to vibrational or light energy and
the electron returns to its ground state. Vibrational energy is transferred through
movement and collision with other molecules, but energy not lost in this way is
released as light radiation. The light emission is known as fluorescence and if some
energy has been removed through vibration, it will be of a lower energy and longer
wavelength than the excitation energy. The wavelength and intensity of the emitted
radiation is dependent on the structure and composition of the molecule and the
excitation wavelength used.

Relationship between concentration and fluorescence

The fluorescence signal F and concentration C of the matter under investigation, are
related by:

ε

-

F = KQP

(1-10

0

bC

)

Where

K = A constant characteristic of the instrument (Including instrument

electronics, pH and Temperature)
Q = Quantum efficiency (= Photons emitted/Photons absorbed)
P

= Power of incident radiation

0

ε = Molar absorptivity of the species (matter)

b = Absorption path length

If the concentration of the matter in question is low (dilute),
relationship is then linear and the equation can be written as

F = 2.3KQP

εbC

0

The accuracy of fluorescent measurements is very high because the radiant energy
being formed is measured directly. There are also only a few, easily controlled limits
on the sensitivity of fluorescence measurements. From the equation above it can be
seen that adjustments made to instrument electrical noise and competing radiations,
along with physical limitations such as radiation energy, sample volume and cell size
affect the measurement sensitivity.

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εbC is small. The