Horiba Scientific FluoroMax-4 User manual
FluoroMax®-4 & FluoroMax®-4P with USB rev. D (30 Jul 2012)
ii
Copyright © 2011, 2012 by HORIBA Instruments Incorporated. All rights reserved. No
part of this work may be reproduced, stored, in a retrieval system, or transmitted in any
form by any means, including electronic or mechanical, photocopying and recording,
without prior written permission from HORIBA Instruments Incorporated. Requests for
permission should be requested in writing. Origin®is a registered trademark of
OriginLab Corporation. Alconox®is a registered trademark of Alconox, Inc. Ludox®is
a registered trademark of W.R. Grace and Co. Teflon®is a registered trademark of E.I.
du Pont de Nemours and Company. Windows®is a trademark of Microsoft Corpora-
tion. Uniblitz®is a registered trademark of VA, Inc.
Information in this manual is subject to change without notice, and does not represent a
commitment on the part of the vendor.
July 2012
Part Number J810005
FluoroMax®-4 & FluoroMax®-4P with USB rev. D (30 Jul 2012)
iii
Table of Contents
0: Introduction ........................................................................................... 0-1
About the FluoroMax®-4 and FluoroMax®-4P ..................................................................................0-1
Chapter overview .............................................................................................................................0-2
Disclaimer.........................................................................................................................................0-3
Safety summary ...............................................................................................................................0-5
Risks of ultraviolet exposure ............................................................................................................0-7
Additional risks of xenon lamps........................................................................................................0-9
CE compliance statement..............................................................................................................0-11
1: Requirements & Installation ........................................................................ 1-1
Safety-training requirements............................................................................................................1-1
Surface requirements.......................................................................................................................1-2
Environmental requirements............................................................................................................1-3
Electrical requirements.....................................................................................................................1-4
Unpacking and Installation...............................................................................................................1-5
Software emulation ..........................................................................................................................1-9
2: System Description ................................................................................... 2-1
Introduction...................................................................................................................................... 2-1
Basic theory of operation .................................................................................................................2-1
Optical layout....................................................................................................................................2-2
3: System Operation...................................................................................... 3-1
Introduction.......................................................................................................................................3-1
Power switch....................................................................................................................................3-1
Turning on the system......................................................................................................................3-2
Checking system performance.........................................................................................................3-3
4: Data Acquisition ....................................................................................... 4-1
Introduction to FluorEssence™ and Multigroup ...............................................................................4-1
Experiment Menu button..................................................................................................................4-2
Previous Experiment Setup button ..................................................................................................4-4
Auto Run Previous Experiment button.............................................................................................4-5
Make Overlay File button.................................................................................................................4-6
3D Scan to 3D Profile button............................................................................................................4-7
Run JY Batch Experiments button...................................................................................................4-9
Real Time Control ..........................................................................................................................4-11
Create/Use Calibration Curve from CWA Data..............................................................................4-12
2D Intensity Map ............................................................................................................................4-16
Launch DataStation........................................................................................................................4-18
Switch menu between HJY Software Application and Origin Std..................................................4-19
Multigroup.......................................................................................................................................4-20
Running an unknown sample.........................................................................................................4-23
Using corrected signals in FluorEssence™ ...................................................................................4-28
5: Optimizing Data....................................................................................... 5-1
Cuvette preparation..........................................................................................................................5-1
Sample preparation..........................................................................................................................5-2
Running a scan on a sample............................................................................................................5-4
Measuring the Gfactor.....................................................................................................................5-5
Improving the signal-to-noise ratio...................................................................................................5-7
Correcting data...............................................................................................................................5-14
6: Maintenance............................................................................................ 6-1
Introduction.......................................................................................................................................6-1
Lamp replacement ...........................................................................................................................6-1
Electronics......................................................................................................................................6-11
Updating the FluoroMax®-4 firmware.............................................................................................6-13
FluoroMax®-4 & FluoroMax®-4P with USB rev. D (30 Jul 2012)
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7: Troubleshooting........................................................................................ 7-1
Chart.................................................................................................................................................7-1
Using diagnostic spectra..................................................................................................................7-4
Further assistance… .......................................................................................................................7-9
8: Producing Correction Factors ....................................................................... 8-1
Introduction.......................................................................................................................................8-1
Generating emission correction factors ...........................................................................................8-2
Calculating emission correction factors ...........................................................................................8-3
Using correction-factor files............................................................................................................8-10
Excitation correction factors...........................................................................................................8-13
9: FluoroMax®-4P Phosphorimeter Operation ....................................................... 9-1
Introduction...................................................................................................................................... 9-1
Theory of operation..........................................................................................................................9-2
Applications for the phosphorimeter ................................................................................................9-5
Operation of the phosphorimeter .................................................................................................... 9-8
Processing phosphorimeter data ...................................................................................................9-10
Lamp replacement .........................................................................................................................9-11
10: Automated Polarizers...............................................................................10-1
Introduction.....................................................................................................................................10-1
Installation......................................................................................................................................10-5
Alignment .......................................................................................................................................10-6
Using automated polarizers .........................................................................................................10-13
Maintenance.................................................................................................................................10-18
Troubleshooting ...........................................................................................................................10-19
11: Technical Specifications ..........................................................................11-1
Introduction.....................................................................................................................................11-1
Spectrofluorometer system............................................................................................................11-2
Minimum host-computer requirements ..........................................................................................11-4
Software.........................................................................................................................................11-4
12: Components & Accessories .......................................................................12-1
Itemized list of FluoroMax®-4 accessories.....................................................................................12-2
FL-1013 Liquid Nitrogen Dewar Assembly ....................................................................................12-3
Sample cells...................................................................................................................................12-4
F-3026 Correction Factor Kit..........................................................................................................12-6
F-3023 Janis cryostat.....................................................................................................................12-7
F4-3000 Fiber Optic Mount and 1950 Fiber Optic Bundles ...........................................................12-8
1939 Cut-On Filter..........................................................................................................................12-9
FM-2008 Cut-On Filter Holder......................................................................................................12-10
FL4-1011 Four-Position Thermostatted Cell Holder....................................................................12-11
FL4-1012 Dual-Position Thermostatted Cell Holder....................................................................12-13
FL4-1027 Single-Position Thermostatted Cell Holder .................................................................12-15
J1933 Solid Sample Holder..........................................................................................................12-17
F-3005/6 Autotitration Injector......................................................................................................12-19
1905-OFR 150-W Xenon Lamp ...................................................................................................12-20
F-3004 Sample Heater/Cooler Peltier Thermocouple Drive ........................................................12-21
MicroMax 384 Microwell Plate Reader ........................................................................................12-22
FM4-2000 L-Format Polarizer......................................................................................................12-23
FL4-1015 Injector Port .................................................................................................................12-24
F-3029 Quanta-φaccessory........................................................................................................12-25
FM-2002 Shutter accessory.........................................................................................................12-26
F-3025 Stopped-flow accessory .................................................................................................12-27
FM4-2015 and FM4-2016 TCSPC upgrade.................................................................................12-28
F4-3030 Temperature Bath..........................................................................................................12-29
F-3031 Transmission Accessory..................................................................................................12-30
J400981 Trigger Cable.................................................................................................................12-31
FM-2007 Windows for the FluoroMax®-4 sample compartment..................................................12-32
13: Glossary ............................................................................................. 13-1
FluoroMax®-4 & FluoroMax®-4P with USB rev. D (30 Jul 2012)
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14: Bibliography .........................................................................................14-1
15: Compliance Information ...........................................................................15-1
Declaration of Conformity...............................................................................................................15-1
Supplementary Information............................................................................................................15-1
16: Index ..................................................................................................16-1
FluoroMax®-4 & FluoroMax®-4P with USB rev. D (30 Jul 2012)
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FluoroMax®-4 & FluoroMax®-4P with USB rev. D (30 Jul 2012) Introduction
0-1
Note:
Keep this and the other reference manuals near the system.
0: Introduction
About the FluoroMax®-4 and FluoroMax®-
4P
Both the FluoroMax®-4 and FluoroMax®-4P are self-contained, fully automated
spectrofluorometer systems. Data output is viewed on a PC, while printouts may be ob-
tained via an optional plotter or printer. All FluoroMax®-4 and FluoroMax®-4P func-
tions are under the control of FluorEssence™ spectroscopy software. The main parts of
the FluoroMax®-4 and FluoroMax®-4P spectrofluorometer systems are:
State-of-the-art optical components
A personal computer
FluorEssence™ for Windows®, the driving software.
The difference between the FluoroMax®-4 and FluoroMax®-4P is that the FluoroMax®-
4P contains a phosphorimeter for phosphorescence measurements. This manual ex-
plains how to operate and maintain a FluoroMax®-4 and FluoroMax®-4P
spectrofluorometer. The manual also describes measurements and tests essential to ob-
tain accurate data. For a complete discussion of the almost limitless power provided by
FluorEssence™, refer to the FluorEssence™ User’s Guide (especially regarding soft-
ware installation) and the on-line help for Origin®and FluorEssence™, which accom-
pany the system.
FluoroMax®-4 & FluoroMax®-4P with USB rev. D (30 Jul 2012) Introduction
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Chapter overview
1: Requirements & Installation
Power and environmental requirements; select the best spot
for the instrument.
2: System Description
How the FluoroMax®-4 and -4P work.
3: System Operation
Operation of the spectrofluorometer system, and calibration
instructions.
4: Data Acquisition
How to use the special FluorEssence™ buttons to acquire
and plot data; how to determine peaks in an unknown sam-
ple.
5: Optimizing Data
Hints for improving the signal-to-noise ratio, instructions
for obtaining corrected data, and other information useful
for optimizing data and ensuring reproducibility.
6: Maintenance
Routine maintenance procedures such as replacing the lamp.
7: Troubleshooting
Potential sources of problems, their most probable causes,
and possible solutions.
8: Producing Correction Factors
How to correct for variation in the system’s sensitivity
across the spectral range.
9: FluoroMax®-4P
Phosphorimeter Operation
Theory, operation, applications, and troubleshooting of the
phosphorimeter, available only on the FluoroMax®-4P sys-
tem.
10: Automated Polarizers
Installation, operation, and troubleshooting of the optional
automated polarizers.
11: Technical Specifications
Instrument specifications and computer requirements.
12: Components & Accessories
Accessories available for the FluoroMax®-4, and how to use
them.
13: Glossary
Some useful technical terms related to fluorescence spec-
troscopy.
14: Bibliography
Other important sources of information.
15: Declaration of Conformity
16: Index
FluoroMax®-4 & FluoroMax®-4P with USB rev. D (30 Jul 2012) Introduction
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Disclaimer
By setting up or starting to use any HORIBA Instruments Incorporated product, you are
accepting the following terms:
You are responsible for understanding the information contained in this document. You
should not rely on this information as absolute or all-encompassing; there may be local
issues (in your environment) not addressed in this document that you may need to ad-
dress, and there may be issues or procedures discussed that may not apply to your situa-
tion.
If you do not follow the instructions or procedures contained in this document, you are
responsible for yourself and your actions and all resulting consequences. If you rely on
the information contained in this document, you are responsible for:
Adhering to safety procedures
Following all precautions
Referring to additional safety documentation, such as Material Safety Data Sheets
(MSDS), when advised
As a condition of purchase, you agree to use safe operating procedures in the use of all
products supplied by HORIBA Instruments Incorporated, including those specified in
the MSDS provided with any chemicals and all warning and cautionary notices, and to
use all safety devices and guards when operating equipment. You agree to indemnify
and hold HORIBA Instruments Incorporated harmless from any liability or obligation
arising from your use or misuse of any such products, including, without limitation, to
persons injured directly or indirectly in connection with your use or operation of the
products. The foregoing indemnification shall in no event be deemed to have expanded
HORIBA Instruments Incorporated’s liability for the products.
HORIBA Instruments Incorporated products are not intended for any general cosmetic,
drug, food, or household application, but may be used for analytical measurements or
research in these fields. A condition of HORIBA Instruments Incorporated’s ac-
ceptance of a purchase order is that only qualified individuals, trained and familiar with
procedures suitable for the products ordered, will handle them. Training and mainte-
nance procedures may be purchased from HORIBA Instruments Incorporated at an ad-
ditional cost. HORIBA Instruments Incorporated cannot be held responsible for actions
your employer or contractor may take without proper training.
Due to HORIBA Instruments Incorporated’s efforts to continuously improve our prod-
ucts, all specifications, dimensions, internal workings, and operating procedures are
subject to change without notice. All specifications and measurements are approximate,
based on a standard configuration; results may vary with the application and environ-
ment. Any software manufactured by HORIBA Instruments Incorporated is also under
constant development and subject to change without notice.
Any warranties and remedies with respect to our products are limited to those provided
in writing as to a particular product. In no event shall HORIBA Instruments Incorpo-
FluoroMax®-4 & FluoroMax®-4P with USB rev. D (30 Jul 2012) Introduction
0-4
rated be held liable for any special, incidental, indirect or consequential damages of any
kind, or any damages whatsoever resulting from loss of use, loss of data, or loss of
profits, arising out of or in connection with our products or the use or possession there-
of. HORIBA Instruments Incorporated is also in no event liable for damages on any
theory of liability arising out of, or in connection with, the use or performance of our
hardware or software, regardless of whether you have been advised of the possibility of
damage.
FluoroMax®-4 & FluoroMax®-4P with USB rev. D (30 Jul 2012) Introduction
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Safety summary
The following general safety precautions must be observed during all phases of opera-
tion of this instrument. Failure to comply with these precautions or with specific warn-
ings elsewhere in this manual violates safety standards of design, manufacture and in-
tended use of instrument. HORIBA Instruments Incorporated assumes no liability for
the customer’s failure to comply with these requirements. Certain symbols are used
throughout the text for special conditions when operating the instruments:
A WARNING notice denotes a hazard. It calls at-
tention to an operating procedure, practice, or sim-
ilar that, if incorrectly performed or adhered to,
could result in personal injury or death. Do not
proceed beyond a WARNING notice until the in-
dicated conditions are fully understood and met.
HORIBA Instruments Incorporated is not respon-
sible for damage arising out of improper use of the
equipment.
A CAUTION notice denotes a hazard. It calls at-
tention to an operating procedure, practice, or sim-
ilar that, if incorrectly performed or adhered to,
could result in damage to the product. Do not pro-
ceed beyond a CAUTION notice until the indicat-
ed conditions are fully understood and met.
HORIBA Instruments Incorporated is not respon-
sible for damage arising out of improper use of the
equipment.
Ultraviolet light! Wear protective goggles, full-
face shield, skin-protection clothing, and UV-
blocking gloves. Do not stare into light.
Intense ultraviolet, visible, or infrared light! Wear
light-protective goggles, full-face shield, skin-
protection clothing, and light-blocking gloves. Do
not stare into light.
Extreme cold! Cryogenic materials must always be
handled with care. Wear protective goggles, full-
face shield, skin-protection clothing, and insulated
gloves.
Explosion hazard! Wear explosion-proof goggles,
full-face shield, skin-protection clothing, and pro-
tective gloves.
Caution:
Caution:
Caution:
Caution:
Caution:
Warning:
FluoroMax®-4 & FluoroMax®-4P with USB rev. D (30 Jul 2012) Introduction
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Risk of electric shock! This symbol warns the user
that un-insulated voltage within the unit may have
sufficient magnitude to cause electric shock.
Danger to fingers! This symbol warns the user that
the equipment is heavy, and can crush or injure the
hand if precautions are not taken.
This symbol cautions the user that excessive hu-
midity, if present, can damage certain equipment.
Hot! This symbol warns the user that hot equip-
ment may be present, and could create a risk of
fire or burns.
Read this manual before using or servicing the in-
strument.
Wear protective gloves.
Wear appropriate safety goggles to protect the
eyes.
Wear an appropriate face-shield to protect the
face.
General information is given concerning operation
of the equipment.
Note:
Caution:
Caution:
Caution:
Caution:
FluoroMax®-4 & FluoroMax®-4P with USB rev. D (30 Jul 2012) Introduction
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Risks of ultraviolet exposure
Do not aim the UV light at anyone.
Do not look directly into the light.
Always wear protective goggles, full-face shield and skin protection
clothing and gloves when using the light source.
Light is subdivided into visible light, ranging from 400 nm (violet) to 700 nm (red);
longer infrared, “above red” or > 700nm, also called heat; and shorter ultraviolet
radiation (UVR), “below violet” or < 400nm. UVR is further subdivided into UV-A
or near-UV (320–400 nm), also called black (invisible) light; UV-B or mid-UV
(290–320 nm), which is more skin penetrating; and UV-C or far-UV (< 290 nm).
Health effects of exposure to UV light are familiar to anyone who has had sunburn.
However, the UV light level around some UV equipment greatly exceeds the level
found in nature. Acute (short-term) effects include redness or ulceration of the skin.
At high levels of exposure, these burns can be serious. For chronic exposures, there
is also a cumulative risk of harm. This risk depends upon the amount of exposure
during your lifetime. The long-term risks for large cumulative exposure include
premature aging of the skin, wrinkles and, most seriously, skin cancer and cataract.
Damage to vision is likely following exposure to high-intensity UV radiation. In
adults, more than 99% of UV radiation is absorbed by the anterior structures of the
eye. UVR can contribute to the development of age-related cataract, pterygium,
photodermatitis, and cancer of the skin around the eye. It may also contribute to
age-related macular degeneration. Like the skin, the covering of the eye or the cor-
nea, is epithelial tissue. The danger to the eye is enhanced by the fact that light can
enter from all angles around the eye and not only in the direction of vision. This is
especially true while working in a dark environment, as the pupil is wide open. The
Caution:
This instrument is used in conjunction with ultra-
violet light. Exposure to these radiations, even reflected
or diffused, can result in serious, and sometimes irre-
versible, eye and skin injuries.
Overexposure to ultraviolet rays threatens human health
by causing:
Immediate painful sunburn
Skin cancer
Eye damage
Immune-system suppression
Premature aging
FluoroMax®-4 & FluoroMax®-4P with USB rev. D (30 Jul 2012) Introduction
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lens can also be damaged, but because the cornea acts as a filter, the chances are re-
duced. This should not lessen the concern over lens damage however, because cata-
racts are the direct result of lens damage.
Burns to the eyes are usually more painful and serious than a burn to the skin. Make
sure your eye protection is appropriate for this work. NORMAL EYEGLASSES OR
CONTACTS OFFER VERY LIMITED PROTECTION!
Training
For the use of UV sources, new users must be trained by another member of the labora-
tory who, in the opinion of the member of staff in charge of the department, is suffi-
ciently competent to give instruction on the correct procedure. Newly trained users
should be overseen for some time by a competent person.
Caution:
UV exposures are not immediately felt. The us-
er may not realize the hazard until it is too late and the
damage is done.
FluoroMax®-4 & FluoroMax®-4P with USB rev. D (30 Jul 2012) Introduction
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Warning:
Xenon lamps
are dangerous.
Please read the fol-
lowing precautions.
Additional risks of xenon lamps
Among the dangers associated with xenon lamps
are:
Burns caused by contact with a hot xenon lamp.
Fire ignited by hot xenon lamp.
Interaction of other nearby chemicals with intense ultraviolet, visible, or infrared
radiation.
Damage caused to apparatus placed close to the xenon lamp.
Explosion or mechanical failure of the xenon lamp.
Visible radiation
Any very bright visible light source will cause a human aversion response: we either
blink or turn our head away. Although we may see a retinal afterimage (which can last
for several minutes), the aversion response time (about 0.25 seconds) normally protects
our vision. This aversion response should be trusted and obeyed. NEVER STARE AT
ANY BRIGHT LIGHT-SOURCE FOR AN EXTENDED PERIOD. Overriding the
aversion response by forcing yourself to look at a bright light-source may result in per-
manent injury to the retina. This type of injury can occur during a single prolonged ex-
posure. Excessive exposure to visible light can result in skin and eye damage.
Visible light sources that are not bright enough to cause retinal burns are not necessari-
ly safe to view for an extended period. In fact, any sufficiently bright visible light
source viewed for an extended period will eventually cause degradation of both night
and color vision. Appropriate protective filters are needed for any light source that
causes viewing discomfort when viewed for an extended period of time. For these rea-
sons, prolonged viewing of bright light sources should be limited by the use of appro-
priate filters.
The blue-light wavelengths (400–500 nm) present a unique hazard to the retina by
causing photochemical effects similar to those found in UV-radiation exposure.
Infrared radiation
Infrared (or heat) radiation is defined as having a wavelength between 780 nm and 1
mm. Specific biological effectiveness “bands” have been defined by the CIE (Commis-
sion Internationale de l’Eclairage or International Commission on Illumination) as fol-
lows:
• IR-A (near IR) (780–1400 nm)
• IR-B (mid IR) (1400–3000 nm)
• IR-C (far IR) (3000 nm–1 mm)
FluoroMax®-4 & FluoroMax®-4P with USB rev. D (30 Jul 2012) Introduction
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The skin and eyes absorb infrared radiation (IR) as heat. Workers normally notice ex-
cessive exposure through heat sensation and pain. Infrared radiation in the IR-A that
enters the human eye will reach (and can be focused upon) the sensitive cells of the ret-
ina. For high irradiance sources in the IR-A, the retina is the part of the eye that is at
risk. For sources in the IR-B and IR-C, both the skin and the cornea may be at risk from
“flash burns.” In addition, the heat deposited in the cornea may be conducted to the lens
of the eye. This heating of the lens is believed to be the cause of so called “glassblow-
ers’ ” cataracts because the heat transfer may cause clouding of the lens.
Retinal IR Hazards (780 to 1400 nm): possible retinal lesions from acute high irra-
diance exposures to small dimension sources.
Lens IR Hazards (1400 to 1900 nm): possible cataract induction from chronic lower
irradiance exposures.
Corneal IR Hazards (1900 nm to 1 mm): possible flashburns from acute high irradi-
ance exposures.
Who is likely to be injured? The user and anyone exposed to the radiation or xenon
lamp shards as a result of faulty procedures. Injuries may be slight to severe.
FluoroMax®-4 & FluoroMax®-4P with USB rev. D (30 Jul 2012) Introduction
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CE compliance statement
The FluoroMax®-4 spectrofluorometer is tested for compliance with both the EMC Di-
rective 89/336/EEC and the Low Voltage Directive for Safety 73/23/EEC, and bears
the international CE mark as indication of this compliance. HORIBA Instruments In-
corporated guarantees the product line’s CE compliance only when original HORIBA
Instruments Incorporated supplied parts are used. Chapter 15 herein provides a table of
all CE Compliance tests and standards used to qualify this product.
FluoroMax®-4 & FluoroMax®-4P with USB rev. D (30 Jul 2012) Introduction
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This manual suits for next models
1
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