Promega QuantiFluor E6090 User manual

Technical Manual
QuantiFluor™
Handheld Fluorometers
Operating Instructions
INSTRUCTIONS FOR USE OF PRODUCTS E6090, E6105 AND E6100.
PRINTED IN USA, Revised 1/11 Part# TM338
8772TA

Promega Corporation · 2800 Woods Hollow Road ·Madison, WI 53711-5399 USA ·Toll Free in USA 800-356-9526 ·Phone [phone] ·Fax [phone] · www.promega.com
Printed in USA. Part# TM338
Revised 1/11 Page 1
1. Introduction...................................................................................................................................................................2
A. Description...........................................................................................................................................................2
B. Product Components .........................................................................................................................................2
C. QuantiFluor™ Handheld Fluorometers General Specifications .................................................................4
D. QuantiFluor™ Handheld Fluorometers Optical Specifications ..................................................................4
E. Inspection.............................................................................................................................................................4
F. Precautions and Special Instructions...............................................................................................................5
G. Environmental Requirements ...........................................................................................................................7
2. Setting up the QuantiFluor™ Handheld Fluorometers .......................................................................................7
3. Instrument Calibration and Operation....................................................................................................................9
A. Instrument Power Up and Shut Down ...........................................................................................................9
B. LCD .......................................................................................................................................................................9
C. Detection Channel Configuration ....................................................................................................................9
D. Calibration Overview.......................................................................................................................................10
E. Calibration .........................................................................................................................................................10
F. Setting Calibration Standard Value ...............................................................................................................11
G. Performing the Calibration .............................................................................................................................11
H. Sample Analysis................................................................................................................................................12
I. QuantiFluor™ PCR Tube Adapter Operation .............................................................................................13
J. Diagnostic Information ....................................................................................................................................13
K. Solid Standard ...................................................................................................................................................14
4. Internal Data Logging (IDL) ....................................................................................................................................15
A. Activating IDL...................................................................................................................................................15
B. Downloading Data to a Computer ................................................................................................................15
C. Erasing Data from the QuantiFluor™ Instrument ......................................................................................15
5. General Considerations ............................................................................................................................................16
A. Handling Samples.............................................................................................................................................16
B. Linear Range and Quenching .........................................................................................................................16
C. Temperature ......................................................................................................................................................16
D. Sample Position.................................................................................................................................................17
E. Data Quality ......................................................................................................................................................17
F. Periodic Cleaning and Maintenance..............................................................................................................17
6. Troubleshooting .........................................................................................................................................................18
7. Internal Data Logging (IDL) ....................................................................................................................................20
A. Shipping Checklist............................................................................................................................................20
B. Hardware Requirements for IDL ...................................................................................................................20
C. Installing IDL Software....................................................................................................................................20
D. Connecting the QuantiFluor™ Fluorometer ................................................................................................20
E. Troubleshooting Internal Data Logging........................................................................................................21
All technical literature is available on the Internet at: www.promega.com/tbs/
Please visit the web site to verify that you are using the most current version of this Technical Manual.
Please contact Promega Technical Services if you have questions on the use of this product. Email: [email protected]
QuantiFluor™ Handheld Fluorometers
Operating Instructions

Promega Corporation · 2800 Woods Hollow Road ·Madison, WI 53711-5399 USA ·Toll Free in USA 800-356-9526 ·Phone [phone] ·Fax [phone] · www.promega.com
Part# TM338 Printed in USA.
Page 2 Revised 1/11
8. Appendix......................................................................................................................................................................22
A. QuantiFluor™ Fluorometer Workflow Chart ..............................................................................................22
B. Returning the QuantiFluor™ Handheld Fluorometer................................................................................24
C. Instrument Disposal .........................................................................................................................................24
D. Certificate of Decontamination.......................................................................................................................25
E. Warranty Information......................................................................................................................................26
F. Service Agreements and Related Products...................................................................................................27
1. Introduction
1.A. Description
The QuantiFluor™ Handheld Fluorometer is a dual-channel fluorometer designed for fast, reliable and
accurate fluorescence measurements. When properly calibrated with a known standard, the
QuantiFluor™ Handheld Fluorometer can be used to determine concentrations of unknown samples.
QuantiFluor™ Handheld Fluorometer Features:
• Compliant with the following EU directives:
•2004/108/EC Electromagnetic Compatibility
•2006/95/EC Low Voltage Directive
• Comprehensive technical support
1.B. Product Components
Product Quantity Cat.#
QuantiFluor™-ST Handheld Fluorometer 1 each E6090
For Research Use. E6090 Filters: Channel A UV 365nm
Ex
/440–470nm
Em
; Channel B Blue (460nm
Ex
, 515–575nm
Em
).
Includes:
• 1 QuantiFluor™ Handheld Fluorometer
• 1 AC Adapter with Interchangeable Plugs
• 4 10 × 10mm Square Methacrylate Cuvettes
• 1 Internal Data Logging (IDL) Package
• 1 RS-232 Cable and Software Instructions
• 1 Spreadsheet Interface Software (1 CD)
• 1 Setup Guide
• 1 CD containing Technical Manual
Available Separately
Product Quantity Cat.#
QuantiFluor™-ST Minicell Adapter Kit 1 each E6112
For measuring 50–250µl of sample. Includes 400 glass cuvettes.
Product Quantity Cat.#
Solid Standard (for use with QuantiFluor™-ST instrument only) 1 each E6113
AC Adapter with Interchangeable Plugs 1 each E6096
QuantiFluor™ PCR Tube Adapter 1 each E6101

Promega Corporation · 2800 Woods Hollow Road ·Madison, WI 53711-5399 USA ·Toll Free in USA 800-356-9526 ·Phone [phone] ·Fax [phone] · www.promega.com
Printed in USA. Part# TM338
Revised 1/11 Page 3
Product Quantity Cat.#
QuantiFluor™-P Handheld Fluorometer (UV/Blue) 1 each E6105
For Research Use. E6105 Filters: Channel A UV 365nm
Ex
/440–470nm
Em
; Channel B Blue (460nm
Ex
, 515–575nm
Em
).
Includes:
• 1 QuantiFluor™ Handheld Fluorometer
• 4 AAA Batteries
• 4 10 × 10mm Square Methacrylate Cuvettes
• 1 Internal Data Logging (IDL) Package
• 1 RS-232 Cable and Software Instructions
• 1 Spreadsheet Interface Software (1 CD)
• 1 Setup Guide
• 1 CD containing Technical Manual
Product Quantity Cat.#
QuantiFluor™-P Handheld Fluorometer (Green/Blue) 1 each E6100
For Research Use. E6600 Filters: Channel A Green 525nm
Ex
/>570nm
Em
; Channel B Blue (460nm
Ex
, 515–575nm
Em
).
Includes:
• 1 QuantiFluor™ Handheld Fluorometer
• 4 AAA Batteries
• 4 10 × 10mm Square Methacrylate Cuvettes
• 1 Internal Data Logging (IDL) Package
• 1 RS-232 Cable and Software Instructions
• 1 Spreadsheet Interface Software (1 CD)
• 1 Setup Guide
• 1 CD containing Technical Manual
Available Separately
Product Quantity Cat.#
QuantiFluor™-P Minicell Adapter Kit 1 each E6111
For measuring 75–250µl of sample. Includes 400 glass cuvettes
Product Quantity Cat.#
QuantiFluor™ PCR Tube Adapter 1 each E6101
See the Related Products Section at the end of this Manual for additional accessories.

Promega Corporation · 2800 Woods Hollow Road ·Madison, WI 53711-5399 USA ·Toll Free in USA 800-356-9526 ·Phone [phone] ·Fax [phone] · www.promega.com
Part# TM338 Printed in USA.
Page 4 Revised 1/11
1.C. QuantiFluor™ Handheld Fluorometers General Specifications
Size QuantiFluor™-ST: 2.7” × 5.5” × 7.25” (6.9 × 14 × 18.42cm)
QuantiFluor™-P: 1.75” x 3.5” x 7.25” (4.45 × 8.9 × 18.4 cm)
Weight QuantiFluor™-ST: 28.8oz. (0.67kg)
QuantiFluor™-P: 14oz. (0.4kg)
Linear Dynamic range 4 logs, assay dependent
Resolution 12 bits
LCD Display 2 × 16 characters
Readout Direct concentration
Detectors Photodiodes: measurement capability from 300–1000nm
Calibration Type Single-point
Alarms Low battery (QuantiFluor™-P instruments only), high blank
Cuvette Type 10 × 10mm plastic, Minicell adapter
Warm-Up Time 5 seconds
Automatic Power Down After 3 minutes of inactivity (QuantiFluor™-P instruments only)
Power Requirements QuantiFluor™-ST:
AC Adapter: 100–240V AC, 50/60 Hz., 0.25A
Instrument: 9V DC, 0.3A
QuantiFluor™-P: 4 AAA Batteries
1.D. QuantiFluor™ Handheld Fluorometers Optical Specifications
1.E. Inspection
Upon receiving your QuantiFluor™ Handheld Fluorometer, please inspect the package carefully to
make sure all items are present and undamaged. Save original packaging in case the instrument needs to
be returned for repair or service. All shipments include:
• QuantiFluor™ Fluorometer (Cat.# E6090, E6105, or E6100)
• 4 AAA Batteries (Cat.# E6105 or E6100 only)
• AC Adapter with interchangeable plugs (Cat. # E6090 only)
• 4 10 × 10mm Square Polystyrene Cuvettes (3.5ml capacity)
• Internal Data Logging (IDL) package
• RS-232 Cable
• Spreadsheet Interface Software (1 CD)
• CD containing Technical Manual
Light Source UV LED* Blue LED
Green LED
(QuantiFluor™-P Instrument,
Cat.# E6100 only)
Excitation Optics 365nm 460nm 525nm
Emission Optics 440–470nm 515–575nm >570nm
Limit of
Detection
10ng/ml dsDNA
using Hoechst Dye
33258 <50pM Fluorescein <200pM Rhodamine
* For UV measurements, we recommend use of methacrylate cuvettes.

Promega Corporation · 2800 Woods Hollow Road ·Madison, WI 53711-5399 USA ·Toll Free in USA 800-356-9526 ·Phone [phone] ·Fax [phone] · www.promega.com
Printed in USA. Part# TM338
Revised 1/11 Page 5
1.F. Precautions and Special Instructions
Safety Symbols and Markings
Danger. Hazardous voltage. Risk of electrical
shock.
Danger. Tension dangereuse. Risque de choc
électrique.
Warning. Risk of personal injury to the
operator or a safety hazard to the equipment
or surrounding area.
Avertissement. Risque de préjudice corporel pour
l'opérateur ou d'accident avec l'instrument ou
l'entourage.
It is important to understand and follow all
laws regarding the safe and proper disposal of
electrical instrumentation. Please contact your
local Promega Representative for disposal of
the instrument and power supply. Please
follow your institutional requirements for
disposal of the accessories.
Avertissement. Il est important de comprendre et de
respecter toutes les lois relatives à la destruction sûre
et correcte des appareils électriques. Veuillez contacter
votre représentant Promega local concernant la
destruction de l’appareil. Veuillez respecter les
exigences de votre établissement concernant la
destruction des accessoires.
Equipment protected by double insulation or
reinforced insulation (Cat.# E6090 AC
Adapter).
Equipement protégé par une double isolation ou par
une isolation renforcée (Cat.# E6090 AC Adapter).
Precautions
IMPORTANT SAFETY INSTRUCTIONS. PLEASE SAVE THESE INSTRUCTIONS.
Do not submerge the QuantiFluor™
Fluorometer in water.
Ne pas submerger le QuantiFluor™ Fluorometer
dans l'eau.
If a liquid is accidentally spilled inside the
Sample Compartment, invert the
QuantiFluor™ Fluorometer to drain the
excess liquid. Then wipe the inside area
dry with a clean soft towel or tissue.
En cas de versement involontaire de liquide dans le
compartiment de lecture, inversez le fluorimètre
QuantiFluor™ pour drainer le liquide résiduel.
Ensuite nettoyez et séchez l'intérieur de l'instrument
avec un tissu souple.
Always disconnect the power before
cleaning or performing routine
maintenance.
Il faut toujours éteindre l'instrument avant de le
nettoyer ou d'effectuer de l'entretien de routine.
Always disconnect the AC Adapter from the
power outlet when not in use (Cat.# E6090).
Toujours enlever l'adaptateur AC de la prise
électrique lorsqu'il n'est pas utilisé (Cat.# E6090)

Promega Corporation · 2800 Woods Hollow Road ·Madison, WI 53711-5399 USA ·Toll Free in USA 800-356-9526 ·Phone [phone] ·Fax [phone] · www.promega.com
Part# TM338 Printed in USA.
Page 6 Revised 1/11
Special Instructions
Note: Failure to operate, store and use the QuantiFluor™ Handheld Fluorometers as directed
will invalidate the warranty.
• Changes or modifications to this unit not expressly approved by the party responsible for
compliance could void the user’s authority to operate the equipment.
• Do not use this device in proximity to sources of strong electromagnetic radiation (e.g.,
unshielded intentional RF sources), because these may interfere with the proper operation.
• Do not use this instrument for anything other than its intended use.
• If the equipment is used in a manner other than that specified by Promega, the protection
provided by the equipment may be impaired.
• Do not disassemble the unit.
• The QuantiFluor™ Fluorometer readily accepts methacrylate cuvettes (Cat.# E6093) for
reading in the UV channel or polystyrene (Cat.# E6092) cuvettes for reading in the BLUE or
GREEN channel. Do not use glass or quartz cuvettes. Do not use the Minicell Cuvettes
without the Minicell Adapter.
• The Sample Compartment cannot accept cuvettes with an outer diameter over 12mm. The 10mm
size refers to the internal diameter of the cuvette, so some plastic cuvettes may be too large.
• Do not force oversized cuvettes into the Sample Compartment. This can damage the Sample
Compartment. If the cuvette does not easily fit into the Sample Compartment, use a different
cuvette.
• Use caution around solvents because they may corrode the plastic case of the QuantiFluor™
Fluorometer.
• If a liquid is accidentally spilled inside the Sample Compartment, invert the QuantiFluor™
Fluorometer to drain the excess liquid. Then wipe the inside area dry with a clean soft towel
or tissue.
Always power off the instrument before cleaning. Be sure to unplug QuantiFluor™-ST
fluorometers before cleaning.
• If extra cleaning is needed, use a mild detergent to dampen the towel and wipe.
• Do not submerge the QuantiFluor™ Fluorometer in water.
• Do not expose the QuantiFluor™ Fluorometer to temperatures outside the specified range
since damage to the unit may occur.
• A minimum volume of 2ml in a 10 × 10mm cuvette is required for best results.
• The Minicell Adapter requires a minimum volume of 50µl for the QuantiFluor™-ST
Fluorometer and 75µl for the QuantiFluor™-P Fluorometers and accepts maximum total
volume of 250µl.
• Avoid having any air bubbles in your sample because these may significantly affect the
fluorescence reading.

Promega Corporation · 2800 Woods Hollow Road ·Madison, WI 53711-5399 USA ·Toll Free in USA 800-356-9526 ·Phone [phone] ·Fax [phone] · www.promega.com
Printed in USA. Part# TM338
Revised 1/11 Page 7
1.G. Environmental Requirements
Shipping and Storage conditions for QuantiFluor™ Fluorometers:
5–40°C under noncondensing conditions, up to 75% humidity
Operating conditions for QuantiFluor™ Fluorometers:
15–30°C, up to 75% humidity, height above sea level: up to 2000m
2. Setting Up the QuantiFluor™ Handheld Fluorometers
QuantiFluor™-ST Fluorometer
1. Position the instrument on a flat surface.
2. Choose the appropriate AC plug for your wall outlet. If you need to remove a plug from the adapter,
depress the button immediately underneath the plug and slide the plug off the AC adapter (Figure 1).
Slide the desired AC plug onto the Adapter from the top until it clicks into place.
Figure 1. Placing the plug on the AC Adapter. Depress the button immediately underneath the plug (arrow 1), and
slide the plug off the AC Adapter in the direction of arrow 2. Choose the appropriate plug and slide onto the AC
Adapter from the top down until it clicks into place.
3. Plug the AC into the QuantiFluor™ Fluorometer (Figure 2).
Be sure to use only the AC Adapter provided with your instrument. Powering the instrument with an
unapproved AC Adapter may damage the unit.
4. Plug the AC Adapter into a wall outlet.
Figure 2. Top view of the QuantiFluor™-ST Fluorometer showing the AC Adapter Input.
5. To turn on the QuantiFluor™-ST, press the ON/OFF button. The instrument takes 5 seconds to warm
up. After the warm-up, the QuantiFluor™-ST is ready for operation.
6. To adjust the brightness of the LCD, press the UP ARROW or DOWN ARROW buttons.
8828TA
Serial portAC Adapter input
1
2
9629TA

Promega Corporation · 2800 Woods Hollow Road ·Madison, WI 53711-5399 USA ·Toll Free in USA 800-356-9526 ·Phone [phone] ·Fax [phone] · www.promega.com
Part# TM338 Printed in USA.
Page 8 Revised 1/11
QuantiFluor™-P Fluorometers
1. On the back of the instrument, loosen the screw and pull the screw up to remove the battery panel
(Figure 3).
2. Install the 4 AAA batteries into the appropriate spaces. Replace the battery panel and tighten the
screw. The panel has an o-ring, which creates a water-tight seal. The battery panel may be difficult to
install if there is no lubrication on the o-ring. Use a silicon-based grease to lightly lubricate the o-ring,
if necessary.
Figure 3. Back Panel of the QuantiFluor™-P Fluorometer showing the battery compartment.
Figure 4. Front Panel of the QuantiFluor™-ST Handheld Fluorometer. The control panel of the QuantiFluor™-P
instrument is identical.
8829TA
UV
xx.x
Indicates active
channel.
Fluorescence reading.
A/B button. Press
to select channel.
SET button. Press to
select sample analysis
mode (Discrete or
Continuous).
READ button. Press to
access READ screens.
DOWN ARROW button.
ENTER button.
CAL button. Press to
access CALIBRATION
screens.
DIAG button. Press to
access DIAGNOSTICS
screens.
ON/OFF button. Turns the
instrument on and off.
STD VAL button. Press to
set numeric value for the
standard solution.
DATA button. Press to
access IDL screens
(Internal Data Logging).
ESC button.
HOME screen.
UP ARROW button.
8827TA

Promega Corporation · 2800 Woods Hollow Road ·Madison, WI 53711-5399 USA ·Toll Free in USA 800-356-9526 ·Phone [phone] ·Fax [phone] · www.promega.com
Printed in USA. Part# TM338
Revised 1/11 Page 9
3. Instrument Calibration and Operation
An instrument workflow chart is provided in Section 8.
3.A. Instrument Power Up and Shut Down
To turn on the QuantiFluor™ Fluorometer, press the ON/OFF button. The instrument takes five seconds
to warm up. After the warm-up, the QuantiFluor™ Fluorometer is ready for use. Pressing the ON/OFF
button again will turn off the unit.
QuantiFluor™-P units only: If left idle for three minutes, the unit will automatically turn off to save
battery power.
A set of new batteries can last over 1,000 sample readings. If the batteries have low power or are not
positioned properly, the following warning message will appear: "Batt Level < 20% !! Caution !!"
QuantiFluor™-ST units only: After switching off the unit, unplug the AC Adapter from the wall outlet.
3.B. LCD
The QuantiFluor™ Fluorometer has a backlit, adjustable-brightness LCD (Liquid Crystal Display) for
easier viewing. The contrast may change with temperature and may get lighter as the unit gets colder.
While on the HOME screen, adjust the brightness of the LCD using the UP ARROW and DOWN
ARROW buttons.
3.C. Detection Channel Configuration
The QuantiFluor™ Fluorometers have two fluorescence detection channels. Identify the configuration by
looking at the label on the back side of the QuantiFluor™ Fluorometer. See the Optical Specifications
table in Section 1.D to confirm the correct configuration for your application.
Select the appropriate channel by pressing the A/B button to toggle between the two channels. The display
will show a label in the lower left corner of the HOME screen to identify which channel is activated.
QuantiFluor™-ST Fluorometer (Cat.# E6090)
Channel A UV (365nmEx, 440–470nmEm),
Channel B Blue (460nmEx, 515–575nmEm)
QuantiFluor™-P Fluorometer (UV, Blue; Cat.# E6105)
Channel A UV (365 nmEx, 440–470nmEm),
Channel B Blue (460 nmEx, 515–575nmEm)
QuantiFluor™-P Fluorometer (Green, Blue; Cat.# E6100)
Channel A Green (525nmEx, >570nmEm)
Channel B Blue (460nmEx, 515–575nmEm)

Promega Corporation · 2800 Woods Hollow Road ·Madison, WI 53711-5399 USA ·Toll Free in USA 800-356-9526 ·Phone [phone] ·Fax [phone] · www.promega.com
Part# TM338 Printed in USA.
Page 10 Revised 1/11
3.D. Calibration Overview
The QuantiFluor™ Fluorometer should be calibrated using a primary standard. A primary standard
contains the same fluorescent material that will be measured in the unknown samples. The
QuantiFluor™ Fluorometer will give an actual quantitative concentration reading when a primary
standard of known concentration is used for calibration. The primary standard is either made to a
known concentration, or to a known dilution factor. Ideally, the primary standard and blank samples,
used for calibration, are made with the same buffer in which the tests are performed. The standard and
samples must be in the linear detection range to achieve accurate quantitative results. See the General
Considerations Section (Section 5) for more details about the linear range and sample quenching.
Notes:
1. The Minicell Adapter requires a minimum volume of 50µl for the QuantiFluor™-ST
Fluorometer and 75µl for the QuantiFluor™-P Fluorometer and accepts maximum total
volume of 250µl.
2. The 10 × 10mm square methacrylate or polystyrene cuvettes require a minimum volume of 2ml.
3. Be sure to insert the Minicell Adapter in the correct orientation when a Minicell cuvette is
used. If the BLUE channel is selected, insert the Minicell Adapter into the Sample
Compartment with the tab away from you and the blue label facing you. If the UV channel is
selected, insert the Minicell Adapter into the Sample Compartment with the tab close to you
and the UV label facing you.
4. The sample compartment lid must be closed before the instrument will read your sample.
Figure 5. Inserting the Quantifluor™-ST Minicell Adapter.
5. The Minicell Adapter for the QuantiFluor™-P is multidirectional and can be inserted in any
orientation.
3.E. Calibration
For best accuracy, always calibrate prior to a sample analysis. The QuantiFluor™ Fluorometer will save
the calibration settings for each channel until a new calibration is performed.
If the temperature of the sample or the QuantiFluor™ Fluorometer changes significantly, the readings
may show a small shift. In this case, consider recalibrating. The Solid Standard (Cat.# E6113, which is
available for the QuantiFluor
™ -ST only) is useful for checking the reading stability over time.
Cuvette orientation and cleanliness also can affect accuracy.
8764TA

Promega Corporation · 2800 Woods Hollow Road ·Madison, WI 53711-5399 USA ·Toll Free in USA 800-356-9526 ·Phone [phone] ·Fax [phone] · www.promega.com
Printed in USA. Part# TM338
Revised 1/11 Page 11
3.F. Setting the Calibration Standard Value
These directions define the numeric value for the standard to be read. The units of measurement are set
in µg/L. For example if a Primary Standard has a 50µg/L concentration, then set the value to 50, and the
implied unit of measure will be in µg/L.
1. Press the STD VAL button to view the value of the standard currently set. The screen will
display “Cal Val: XX.X <UP/DOWN> BLUE”.
Note: Be quick. After after 5 seconds, the display defaults to home. This standard value will
apply only to the channel you have previously selected using the A/B button (Section 3.C).
2. Press the UP ARROW or DOWN ARROW buttons to set the standard value. Holding down
either of the arrow buttons will result in faster scrolling of the value.
3. When you have set the standard value, press the ENTER to accept the value and return to
the HOME screen.
3.G. Performing the Calibration
1. Press the CAL button, the screen will display “Calib BLUE <ENT> to start”.
2. Press the ENTER button to move to the next screen, which will display “Insert Blank then
press <ENT>”.
3. Insert the blank sample, and press the ENTER button. The QuantiFluor™ Fluorometer will
average the reading for 10 seconds and set the zero (blank) point. During this time the screen
will display “Reading Blank”.
4. After the instrument has set the zero point, the screen will display “Insert Cal Soln then press
<ENT>”.
5. Insert your standard sample and press the ENTER button.
6. The average reading is taken for 10 seconds, and the Standard Calibration value is set. Once
the value is set, the screen will display “Calib Complete press <ENT>”.
Note: If the instrument was calibrated with the highest concentration standard, and the RFU
of the standard is too close to that of the blank, the instrument will register a
"Standard<Blank Recalibrate" message. The RFU of the standard (RFU-STD) must be at least
3X greater than the RFU of the blank (RFU-BLK) in order to calibrate the instrument. If the
"Standard<Blank Recalibrate" message appears, we recommend increasing the standard
concentration by 10X or examining the blank for high fluorescence background.
7. Press the ENTER button to accept the calibration. If the ENTER button is not pressed within
10 seconds, a message will display asking if you wish to abort the calibration. Press the UP
ARROW to abort the calibration or the DOWN ARROW to accept it. If you abort the
calibration, the display will return tot he HOME screen, and the instrument will default to the
previous calibration.
Note: At any time during Steps 1-4, you can press the ESC button to stop the calibration. The
display will return to the HOME screen, and the instrument will default to the previous
calibration.

Promega Corporation · 2800 Woods Hollow Road ·Madison, WI 53711-5399 USA ·Toll Free in USA 800-356-9526 ·Phone [phone] ·Fax [phone] · www.promega.com
Part# TM338 Printed in USA.
Page 12 Revised 1/11
3.H. Sample Analysis
The QuantiFluor™ Fluorometers are capable of measuring samples in two reading modes: Discrete or
Continuous. In Discrete Mode, the instrument measures and averages the fluorescence signal over a 5-
second interval. In Continuous Mode, the instrument measures and averages the fluorescence signal
over a user-defined interval for each sample measurement. The instrument measures the sample
repeatedly for a user-defined number of samples.
Discrete Mode
1. Press the SET button to set the read mode.
2. Press the UP ARROW or the DOWN ARROW button to toggle between Discrete and
Continuous Modes.
3. Once the screen reads "Read Mode: Discrete", press the ENTER button.
4. Insert the sample. Using the specified cuvette or the Minicell Adapter and Minicell Cuvette.
Note: The sample compartment lid must be closed before the instrument will read your sample.
5. Press the READ button. The instrument will measure and average the fluorescence signal
over a 5-second interval. During this time the instrument screen will indicate "Reading". The
result will be displayed at the top center of the screen.
6. Once the "Reading" message disappears, another sample can be read.
Continuous Mode
Note: For Continuous Mode, either the Internal Data Logging function must be enabled or the
instrument must be connected to an external PC or printer.
1. Press the SET button to set the read mode.
2. Press the UP ARROW or the DOWN ARROW button to toggle between Discrete and
Continuous Modes.
3. Once the screen reads "Read Mode: Continuous", press the ENTER button.
4. The screen will read "Continuous Meas Interval: 5 sec." Use the UP ARROW and DOWN
ARROW buttons to change the interval time.
5. Press the ENTER button when your desired interval time is displayed. (5, 10, 30 or 60 seconds)
6. The screen will display "Cont Meas Count". Use the UP ARROW and DOWN ARROW
buttons to select the number of samples you want to measure. Press the ENTER button.
7. Press the READ button.
8. The screen will display "CONT: count = XX <ENT> int = x sec". Press the ENTER button.
9. The screen will display "Insert Sample Press <ENTER>". Insert your sample, close the compartment
lid, and press the ENTER button again.
Note: The sample compartment lid must be closed before the instrument will read your sample.

Promega Corporation · 2800 Woods Hollow Road ·Madison, WI 53711-5399 USA ·Toll Free in USA 800-356-9526 ·Phone [phone] ·Fax [phone] · www.promega.com
Printed in USA. Part# TM338
Revised 1/11 Page 13
10. During reading the instrument screen will indicate "READING” or “WAITING”. The result
will be displayed at the top, center of the screen when all counts are measured. At this point,
another sample can be read.
11. T
O read next sample repeat steps 7–10.
3.I. QuantiFluor™ PCR tube Adapter Operation
The QuantiFluor™ PCR tube Adapter (Cat.# E6101) allows measurement of small
sample volumes in 0.5 ml PCR tubes, while still preserving the superior sensitivity
of the QuantiFluor™ Fluorometer. The PCR tube Adapter is multidirectional and
can be inserted into the Optical Kit sample compartment in any orientation.
Transfer the sample to a 0.5 ml PCR tube. The minimum volume required for
reproducible results is 100μl. Insert the PCR tube into the PCR tube Adapter, close
the lid, and measure the sample. Take care not to introduce air bubbles into the
sample.
3.J. Diagnostic Information
The Diagnostic screens show the number of points available and the current values for the calibration
and blank standards.
1. Press the DIAG button to access the diagnostic screens.
2. The first screen shows the number of available data points for internal data logging.
3. Press the ENTER button to toggle to the RFU (Relative Fluorescence Unit) values from the
calibration blank and standard.
4. Press the ESC button when finished to return to the HOME screen.
Note: If the instrument was calibrated with the highest concentration standard, and the RFU of
the standard is too close to that of the blank, the instrument will register a Standard<Blank Error
message.The RFU value of the Standard must be 3 times greater than the RFU value of the blank.
9628TA
QuantiFluor™
PCR tube Adapter.

Promega Corporation · 2800 Woods Hollow Road ·Madison, WI 53711-5399 USA ·Toll Free in USA 800-356-9526 ·Phone [phone] ·Fax [phone] · www.promega.com
Part# TM338 Printed in USA.
Page 14 Revised 1/11
3.K. Solid Standard
Note: The Solid Standard (Cat.# E6113) is for use with the QuantiFluor™-ST Handheld Fluorometer
(Cat.# E6090) only.
The Solid Standard (Cat.# E6113) is designed to verify the stability of the QuantiFluor™-ST Handheld
Fluorometer by checking drift of the calibration. It will also verify if any instrument settings have been
changed.
1. After calibration, insert the Solid Standard into the sample compartment of the
QuantiFluor™-ST Fluorometer.
Note: always insert the Solid Standard with the tab away from you.
2. Press the READ button.
3. Record the reading.
4. Check the calibration and/or the stability of the instrument by periodically reading the Solid
Standard.
5. Confirm that the current measured value does not deviate from previously recorded value by
5% or more. If the value of the Solid Standard deviates by 5% or more, it is time to recalibrate
the instrument (see Sections 3.D–3.G)
Note: If you recalibrate the channel, then a new measurement for the Solid Standard must be
taken and stored for future reference.

Promega Corporation · 2800 Woods Hollow Road ·Madison, WI 53711-5399 USA ·Toll Free in USA 800-356-9526 ·Phone [phone] ·Fax [phone] · www.promega.com
Printed in USA. Part# TM338
Revised 1/11 Page 15
4. Internal Data Logging (IDL; additional information is available in Section 7)
4.A. Activating IDL
1. Press the DATA button 2 times.
2. Press the ENTER button to choose between logging and stop.
3. Press the ESC button when finished to return to the HOME screen.
Note: Once activated, the IDL is capable of storing 1000 data points
4.B. Downloading Data to a Computer
Important Note: Before you can download data that have been logged on your QuantiFluor™
Fluorometer to a computer, that computer must have the Spreadsheet Interface Software Installed. See
Section 7 for computer requirements and installation instructions.
1. Connect the QuantiFluor™ Fluorometer to the computer serial port using the provided
interface cable.
2. Open the Spreadsheet Interface Software.
3. Press the DATA button 3 times. The screen will display: "Download data: 5X<ENT> to start".
4. Press the ENTER button 5 times to start downloading the data.
5. Press the ESC button when the data have downloaded to return to the HOME screen.
Note: If no data are logged, the screen will display the following message "Data logger has no
valid data."
4.C. Erasing Data from the QuantiFluor™ Instrument
1. Press the DATA button 4 times. The screen will display "Erase data: 5X <ENT> to start."
2. Press the ENTER button 5 times to erase all logged data from the QuantiFluor™ Instrument. When
the data are erased, the screen will display the following message: "Erase Data All data erased."
3. Press the ESC button to return to the HOME screen.

Promega Corporation · 2800 Woods Hollow Road ·Madison, WI 53711-5399 USA ·Toll Free in USA 800-356-9526 ·Phone [phone] ·Fax [phone] · www.promega.com
Part# TM338 Printed in USA.
Page 16 Revised 1/11
5. General Considerations
5.A. Handling Samples
1. Take care not to spill liquid into the Sample Compartment. Promptly wipe any spills.
2. The QuantiFluor™ Fluorometers are very sensitive, and even small amounts of material from
a previous sample may result in errors. Use a clean cuvette for each reading. Thorough and
proper cleaning of cuvettes between sample readings is essential and is especially important if
the same cuvette is used for both the sample and the blank.
3. The minimum volume is 2ml for 10 × 10mm cuvette, or 50µl and 75µl for the QuantiFluor™-
ST and QuantiFluor™-P Minicell Adapters, respectively.
4. Ensure that the cuvette is clean and dry on the outside when taking readings. Moisture and
condensation on the outside can result in error.
5. Minute bubbles in samples will cause errors in the readings. Be sure not to introduce bubbles
into samples. Particular care must be taken with the Minicell Adapter. Slight tapping on the
outside cuvette wall often will help dissipate bubbles.
5.B. Linear Range and Quenching
The linear range is the concentration range in which the QuantiFluor™ Fluorometer readout is directly
proportional to the concentration of the fluorophore. The linear range begins with the smallest detectable
concentration and continues to an upper concentration limit that depends upon the properties of the
fluorescent material, the filters used, and the path length.
A nonlinear relationship occurs at higher concentrations where the fluorescence signal increases at
decreasing rates in relation to the concentration change. At even higher concentrations, readings begin to
decrease despite increasing sample concentrations. This effect is known as "signal quenching."
Linearity may be checked by diluting a sample 1:1 or some other appropriate ratio (be sure to use a
corresponding buffer for the dilutions). If the sample is within the linear range, the reading will decrease
in direct proportion to the dilution. If the reading does not decrease in direct proportion to the dilution
or if the reading increases, the sample is beyond the linear range of the fluorophore.
5.C. Temperature
Fluorescence is temperature-sensitive. As the temperature of the sample increases, the fluorescence
decreases. For greatest accuracy, read the blank, standard and samples at the same temperature.

Promega Corporation · 2800 Woods Hollow Road ·Madison, WI 53711-5399 USA ·Toll Free in USA 800-356-9526 ·Phone [phone] ·Fax [phone] · www.promega.com
Printed in USA. Part# TM338
Revised 1/11 Page 17
5.D. Sample Position
For low-concentration samples, cuvettes often give slightly different measurements depending upon
their orientation in the sample compartment. This is due to defects in the shape of the cuvette that are
not visible to the human eye. We recommend marking the cuvette at the top and positioning it in the
Sample Compartment the same way each time to minimize error.
5.E. Data Quality
The QuantiFluor™ Fluorometer is only as accurate as the standards that are used to calibrate it. It is
important to take care when preparing standards, samples and blank. Follow the appropriate laboratory
practices when preparing all solutions and reagents.
5.F. Periodic Maintenance and Cleaning
Keep the outside of the instrument clean by wiping with a damp cloth. Remove spills or stains with a
cloth dampened with a mild detergent.
If a liquid is accidentally spilled inside the Sample Compartment, invert the QuantiFluor™ Fluorometer
to drain the excess liquid. Then wipe the inside area dry with a clean soft cloth. If extra cleaning is
needed use a cloth dampened with a mild detergent.
Do not use solvents; they may corrode the plastic case of the QuantiFluor™ Fluorometer.
Do not submerge the QuantiFluor™ Fluorometer in water. Do not disassemble the unit.
Always power off the instrument before cleaning. Be sure to unplug the QuantiFluor™- Fluorometer
before cleaning.

Promega Corporation · 2800 Woods Hollow Road ·Madison, WI 53711-5399 USA ·Toll Free in USA 800-356-9526 ·Phone [phone] ·Fax [phone] · www.promega.com
Part# TM338 Printed in USA.
Page 18 Revised 1/11
6. Troubleshooting
For questions not addressed here, please contact your local Promega Branch Office or Distributor. Contact information available at:
www.promega.com. E-mail: [email protected]
Symptoms Possible Causes and Comments
Low or no reading Use the Solid Standard to determine if the
instrument is functioning properly (with
QuantiFluor™-ST, Cat.# E6090 only; Section 3.K). If
the value of the Solid Standard deviates from the
previous reading by 5% or more, recalibrate the
instrument.
Check to ensure that all reagents and standards
are made to specification and not expired. Make
sure that dilutions were performed correctly.
Protect sample from light. Some fluorophores
are susceptible to photodegradation.
Make sure the correct optical channel is selected.
Make sure that the instrument is properly
calibrated before reading samples.
Ensure that the sample signal is within the linear
range of detection. Perform a dilution series if
necessary.
Ensure that the sample cuvette is appropriate.
Methacrylate cuvettes should be used for UV
applications.
Avoid sudden temperature fluctuations.
Ensure that sample volume is at least 2ml for
the 10 × 10mm cuvette or 50µl for the
QuantiFluor™-ST Minicell cuvette or 75µl
for the QuantiFluor™-P Minicell cuvette.
If you are using a QuantiFluor™-ST Minicell
Adapter, be sure that it is in the correct orientation.
If the BLUE channel is selected, insert the Minicell
Adapter in the Sample Compartment with the tab away
from you and the BLUE label facing you. If the UV
channel is selected, insert the Minicell Adapter in the
Sample Compartment with the tab close to you and the
UV label facing you.

Promega Corporation · 2800 Woods Hollow Road ·Madison, WI 53711-5399 USA ·Toll Free in USA 800-356-9526 ·Phone [phone] ·Fax [phone] · www.promega.com
Printed in USA. Part# TM338
Revised 1/11 Page 19
Symptoms Possible Causes and Comments
Low or no reading (continued) Ensure that the orientation of the Minicell
cuvettes is consistent.
Make sure that the Relative Fluorescence Unit
value of the calibration standard is at least 3X
greater than the blank. If the values of the stan-
dard and the blank are too close, the instrument
will register a Standard<Blank error message.
Readings outside the linear range Dilute samples and standards so that the signal
of detection falls within the linear range of detection.
Fluctuating results Use the Solid Standard to determine
if the instrument is functioning properly.
Check for spills in the sample compartment. If
liquid is spilled inside the sample compartment,
invert the fluorometer to drain the excess liquid.
Wipe the inside area with a clean soft cloth.
Clean any spills with a damp cloth or a towel
dampened with a mild detergent.
Ensure that sample volume is at least 2ml for a
10 × 10mm cuvette or 50µl for the QuantiFluor™-ST
or 75µl QuantiFluor™-P Fluorometer minicell
cuvette, respectively.
Ensure that the sample cuvette is appropriate.
Methacrylate cuvettes should be used for UV
applications.
Avoid dramatic temperature fluctuations.
Fluorescence is temperature-sensitive. As the
temperature of the sample increases, the
fluorescence decreases. For greatest accuracy,
read the blank, standard and samples at the
same temperature.