Turner BioSystems Picofluor User manual

Picofluor™
Handheld Fluorometer
Operating Manual
Part Number: 998-0853
Version: 1.5

Turner BioSystems, Inc. and its suppliers own the written and other materials in the
Picofluor™ Handheld Fluorometer. The written materials are copyrighted and protected by
the copyright laws of the United States of America and, throughout the world, by the
copyright laws of each jurisdiction according to respective treaties and conventions. No part
of this manual may be reproduced manually, electronically, or any other form therein
without the written permission of Turner BioSystems, Inc.
Picofluor™is a trademark of Turner BioSystems, Inc. All of the trademarks, service marks,
and logos used in these materials are the property of Turner BioSystems, Inc. or their
respective owners.
All goods and services sold are subject to the terms and conditions of sale of the company
within the Turner BioSystems, Inc. group that supplies them. A copy of these terms and
conditions is available upon request.
NOTICE TO PURCHASER: The Picofluor™ Handheld Fluorometer is for research
purposes only. It is not intended or approved for diagnosis of disease in humans or
animals.
© Turner BioSystems, Inc. 2007 — All rights reserved.
Made in the USA
Part Number: 998-0853
Version: 1.5
February 9, 2010

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Table of Contents
1Introduction ....................................................................................................................................2
1.1 Description .....................................................................................................................................2
1.2 General Information and Precautions.................................................................... 2
1.3 LCD...................................................................................................................................................2
1.4 Inspection.......................................................................................................................................2
1.5 Setup................................................................................................................................................2
2Quick View Diagrams..................................................................................................................... 3
3Instrument Workflow .....................................................................................................................4
4Instrument Calibration and Operation .........................................................................................6
4.1 Instrument Power Up..................................................................................................................6
4.2LCD...................................................................................................................................................6
4.3Detection Channel Configuration............................................................................................6
4.4Calibration Overview...................................................................................................................6
4.5Calibration ......................................................................................................................................7
4.6Setting Calibration Standard Value........................................................................................7
4.7Performing the Calibration ........................................................................................................7
4.8Sample Analysis...........................................................................................................................7
4.9Diagnostic Information................................................................................................................8
5Internal Data Logging (IDL)...........................................................................................................9
5.1 To activate the IDL.......................................................................................................................9
5.2 Download Data .............................................................................................................................9
5.3 Erase Data .....................................................................................................................................9
6General Considerations for Analysis.........................................................................................10
6.1 Handling Samples......................................................................................................................10
6.2 Linear Range and Quenching................................................................................................10
6.3 Temperature Considerations..................................................................................................10
6.4 Positioning Samples..................................................................................................................11
6.5 Data Quality.................................................................................................................................11
7Warranty........................................................................................................................................12
7.1 Terms.............................................................................................................................................12
7.2 Warranty Service........................................................................................................................12
7.3 Out-of-Warranty Service..........................................................................................................13
APPENDIX A Instrument Specifications..............................................................................................14
A.1 General Specifications .............................................................................................................14
A.2 Optical Specifications ...............................................................................................................14
APPENDIX B Internal Datalogging .......................................................................................................15
B.1 Shipping Checklist.....................................................................................................................15
B.2 Hardware Requirements..........................................................................................................15
B.3 Installation ....................................................................................................................................15
B.4 Connecting...................................................................................................................................15
B.5 Troubleshooting..........................................................................................................................15

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1Introduction
1.1 Description
The Picofluor™ is a dual-channel fluorometer designed for fast, reliable and accurate fluorescence
measurements. When properly calibrated with a known standard, the Picofluor™ can be used to
determine concentrations of unknown samples.
1.2 General Information and Precautions
•The Picofluor™ readily accepts methacrylate or polystyrene cuvettes. Do not use glass or quartz
cuvettes.
•The Sample Compartment cannot accept cuvettes with an outer diameter over 12 mm. The 10 mm
size refers to the internal diameter of the cuvette so some plastic cuvettes may also 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 as they may corrode the plastic case of the Picofluor™.
•If a liquid is accidentally spilled inside the Sample Compartment, invert the Picofluor™ to drain the
excess liquid. Then wipe the inside area dry with a clean soft towel or tissue.
•If extra cleaning is needed, use a mild detergent to dampen the towel and wipe.
•Do not submerge the Picofluor™ in water.
•Do not expose the Picofluor™ to temperatures outside the specified range of 5 oC to 40 oC. Damage
may occur to the unit and will not be covered under warranty.
•A minimum volume of 2 ml in a 10 x 10 mm cuvette is required for best results.
•The Minicell Adapter requires a minimum volume of 75 ml and a maximum volume of 250 ml.
•Avoid having any air bubbles in your sample as they can significantly affect the fluorescence reading.
1.3 Inspection
Upon receiving the instrument, inspect everything carefully to make sure all accessories are present. All
shipments include:
•The Picofluor™
•4 AAA Batteries
•4 Plastic Cuvettes
•Internal Data Logging (IDL) package
•PC Interface Cable and Software Instruction
•Spreadsheet Interface Software (1CD)
•This Operating Manual
1.4 Setup
Before the Picofluor™ can be used; the supplied batteries must be installed.
1. On the back of the instrument, loosen the screw and pull the screw up to remove the battery panel
See Section 2 for a diagram.
2. Install the 4 AAA batteries into the appropriate spaces.
3. Replace the battery panel and tighten the screw. The panel has an o-ring, which creates a watertight
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.

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2Quick View Diagrams
Backlit Screen

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3Instrument Workflow
Turner Designs
TD HH1B 0500 5
5 second warm up
XXX.X
A
XXX.X
A
Calib. A
<ENT> to start READING A
Cal Val: XXX.X
<UP/DOWN> A
XXX.X
B
Insert Blank
then press <ENT>
Reading Blank
XX.X
Insert Cal Soln
then press <ENT>
Reading Cal Soln
XX.X
Calibration
Completed <ENT>
Abort Cal?
<UP>Yes <DOWN>No
WAIT XXX.X
A
XXX.X
A
Logging Points
Left: XXX
%FS-Blk: X.XX
%FS-STD: XX.XX
HOME Screen
Press A/B to toggle
between
and
Indicates which channel
is active Indicates activity of
IDL
Press CAL Press READ
and
If ENT is not pressed
within 5 seconds,
this screen will appear
Fluorescence reading
Press DIAG
then ENTER to
toggle between
Press STD VAL

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Datalogger:
<DATA> to toggle
Status: Stop
<ENT> to toggle
Status: Logging
<ENT> to toggle
Download data:
5X<ENT> to start Erase Data 5X<ENT> to start
Data logger has
no valid data!
Download data:
All downloaded Erase Data All data erased
Press DATA
Press DATA 1X Press DATA 2X
Press ENTER
If there is no data logged, this
screen will appear
Press DATA 3X

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4Instrument Calibration and Operation
4.1 Instrument Power Up
To turn on the Picofluor™, press the ON/OFF button. The instrument takes 5 seconds to warm up. After
the warm-up, the Picofluor™ is ready for use.
Pressing the ON/OFF button again will turn off the unit or, if left idle for 3 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 !!”
4.2 LCD
The PicoFluor™ 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, press the UP ARROW button to darken the contrast and press the DOWN ARROW button to
lighten it.
4.3 Detection Channel Configuration
The Picofluor™ has 2 fluorescence detection channels. Identify the configuration by looking at the label
on the back side of the Picofluor™. See the table in Appendix A to confirm the correct configuration for
your application.
Select the appropriate channel by pressing the A/B button to toggle between the 2 channels. The display
will show a label in the lower left corner of the HOME screen to identify which channel is activated.
Model 8000-003
Channel A
UV
= Ultra Violet excitation
Channel B
Blue
= Blue LED excitation
Model 8000-004
Channel A
RHOD
= Green LED excitation
Channel B
Blue
= Blue LED excitation
See Appendix A for optical (wavelength) specifications.
4.4 Calibration Overview
The Picofluor™ should be calibrated using a Primary Standard. A Primary Standard contains the same
fluorescent material that is measured in the unknown samples. The Picofluor™ 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, typically in ppb or ug/L units,
or to a known dilution factor. Ideally, the Primary Standard and Blank samples, used for calibration, are
made with the same water in which the tests are performed. The standard and samples must be in the
linear detection range to achieve accurate quantitative results. See Section 6.2 for more details about the
linear range and sample quenching.

4. Instrument Calibration and Operation
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4.5 Calibration
For best accuracy, always calibrate prior to a sample analysis. The Picofluor™ will save the calibration
settings for each channel until a new calibration is performed.
If the temperature of the sample or the Picofluor™ changes significantly, the readings may show a small
shift. In this case, consider recalibrating. The solid Secondary Standard is useful for checking the reading
stability over time and can also be used to recalibrate if needed.
Cuvette orientation and cleanliness can have an impact on the result accuracy.
See Section4.8 for sample analysis guidelines to help insure the best results for the analysis.
4.6 Setting Calibration Standard Value
This defines the numeric value for the standard to be read. For example, if a Primary Standard has a 50
ug/L concentration, then set the value to 50 and the implied unit of measure will be in ug/L.
1. Press the STD VAL button.
2. Press the UP and DOWN ARROW buttons to set the standard value. Holding down either arrow
buttons will result in fast scrolling of the value.
3. When finished, press the ENTER or ESC button to accept the value and return to the HOME screen.
4.7 Performing the Calibration
1. Press the CAL button.
2. Press the ENTER button to start the calibration.
3. Insert the blank sample and press the ENTER button. The Picofluor™ will average the reading for 10
seconds and set the blanking zero point.
4. Insert the standard sample and press the ENTER button.
5. The reading is averaged for 10 seconds and the Standard Calibration value is set.
6. Press the ENTER button when calibration is complete to accept the calibration. If the ENTER button
is not pressed within 10 seconds, a dialog will appear asking the user if aborting the calibration is
desired. Press the UP or DOWN ARROW button to abort or accept the calibration, respectively.
At anytime during steps 1-4, press the ESC button to stop the calibration. The display will return to the
HOME screen and will default the instrument to the previous calibration.
4.8 Sample Analysis
1. Insert a minimum of 2 mL of the sample using a plastic cuvette only.
2. Press one of the READ buttons. The instrument will measure and average the fluorescence signal
over a 5-second interval.
3. The result will be displayed at the top and center of the HOME screen.
4. The top left corner will display “WAIT” for 5 seconds. Once the message “WAIT” disappears, the
Picofluor™ is ready to measure another sample.

4. Instrument Calibration and Operation
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4.9 Diagnostic Information
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.
If the instrument is calibrated using the highest standard concentration and the RFU-Std value is too
close to the RFU-Blk value, the sample reading may exceed the instrument’s detection limit.
Ideally: RFU-Std value ≥ 3
RFU-Blk value

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5Internal Data Logging (IDL)
The Picofluor™ can log up to 1,000 data points. The DATA screens control logging, downloading, and
erasing of data.
5.1 To activate the IDL
1. Press the DATA button 2 times.
2. Press the ENTER button to toggle between logging and stopping.
3. Press the ESC button when finished to return to the HOME screen.
5.2 Download Data
1. Connect the Picofluor™ to the PC’s serial port using the provided interface cable.
2. Open the Spreadsheet Interface Software. See Appendix B for computer requirements and
installation.
3. Press the DATA button 3 times.
4. Press the ENTER button 5 times to start the data download.
5. Press the ESC button when finished to return to the HOME screen.
5.3 Erase Data
1. Press the DATA button 4 times.
2. Press the ENTER button 5 times to erase all logged data.
3. Press the ESC button when finished to return to the HOME screen.

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6General Considerations forAnalysis
6.1 Handling Samples
1. Take care not to spill liquid into the Sample Compartment. Promptly wipe any spills.
2. The Picofluor™ is 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. Fill the cuvette at least 50% full. Significant error will result if it is not adequately filled.
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 will often help dissipate bubbles.
6.2 Linear Range and Quenching
The linear range is the concentration range in which the Picofluor™ 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. See the diagram below.
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.
6.3 Temperature Considerations
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.
0
1
2
3
4
5
6
7
0 2 4 6 8 10 12
Fluorophore conc.
Fluorescence

6. General Considerations for Analysis
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6.4 Positioning Samples
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. It is recommended that the cuvette be marked at the top and positioned in the
Sample Compartment the same way each time to minimize error.
6.5 Data Quality
The Picofluor™ 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.

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7Warranty
7.1 Terms
Turner BioSystems warrants the Picofluor™ Fluorometer and any accessories to be free from defects in
materials and workmanship under normal use for a period of one year from the time of initial purchase
under the following restrictions:
•The instrument and accessories must be installed, powered, and operated in compliance with the
directions in this Operating Manual and the directions accompanying the accessories.
•Damage incurred during shipping is not covered by warranty.
•Damage resulting from measuring incompatible samples is not covered by warranty.
•Damage resulting from reagent spills is not covered by warranty.
•Damage resulting from contact with corrosive materials or atmosphere is not covered by warranty.
•Damage from salts, proteins and other moderately corrosive materials that are not promptly removed
from the instrument are not covered by warranty.
•Damage caused by user modification of the instrument is not covered by warranty.
7.2 Warranty Service
To obtain service during the warranty period, take the following steps:
1. Write or call the Turner BioSystems Service Department and describe as precisely as possible the
nature of the problem.
2. Carry out minor adjustments or tests as suggested by the Technical Support Department.
3. If the instrument is still not functioning properly, you must obtain an RMA number from the Turner
BioSystems Service Department before shipping the instrument. This number is necessary for
tracking and repair under warranty.
4. Reference the RMA number on the outside of the package. Pack the instrument well as damage
incurred during shipping due to improper packing is not covered by warranty. Purchase shipping
insurance for the package and ship it to the Turner BioSystems Service Department. All shipments
must be prepaid.
The instrument will be repaired and returned free of charge for any customer in the United States. Turner
BioSystems will pay for return shipment and include a reimbursement check for the cost of surface
shipment to the Service Department.
Turner BioSystems cannot pay shipping, duties, or documentation costs outside the continental United
States. Contact Turner BioSystems for instructions if you are an international customer who made a
purchase directly from Turner BioSystems (not from a third party distributor). Customers outside of the
United States who have purchased our equipment from an authorized distributor should contact their
distributor directly for further instructions. The instrument will be repaired at no charge if it is under
warranty.
IMPORTANT: Under no circumstances should the instrument or accessories be returned without prior
authorization from Turner BioSystems or our authorized distributor.
Prior correspondence is needed to eliminate minor problems easily handled on-site in your laboratory in
order to save money and minimize equipment downtime. Prior correspondence can also help determine
the exact nature of the problem so that repairs can be tailored accordingly.

7. Warranty
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7.3 Out-of-Warranty Service
Follow the same steps as for Warranty Service. Our Service Department is happy to assist you by
telephone or correspondence at no charge. Repair service will be billed at a flat rate. Your invoice will
include freight charges.
Mailing Address:
Turner BioSystems
645 North Mary Avenue
Sunnyvale, CA 94085
USA
Phone: [phone]
Toll-Free: 1(888) 636-2401 (US/Canada)
Fax: [phone]

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APPENDIX A Instrument Specifications
A.1 General Specifications
A.2 Optical Specifications
Specification
Description
Size 1.75” x 3.5” x 7.25”
(4.45 cm x 8.9 cm x 18.4 cm)
Weight 14 oz (0.4 kg)
Dynamic range 4 orders of magnitude
Resolution 12 bits
LCD Display 2 x 16 characters
Case Meets IP 67 Standard; dustproof and waterproof
Temperature 41 - 104 oF; 5 - 40 oC
Detectors Photodiodes: measurement capability from 300—1000 nm
Calibration Type Single-point
Alarms Low battery, circuit failure,
high blank
Cuvette Type 10 x 10 mm plastic
Warm-Up Time 5 seconds
Automatic Power Down After 90 seconds of inactivity
Light Source UV LED Blue LED Green LED
Excitation Optics 365 nm 460 nm 525 nm
Emission Optics 440 - 470 nm 515 - 575 nm >570 nm
Limit of Detection <50 pM Fluorescein <200 pM Rhodamine

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APPENDIX B Internal Datalogging
B.1 Shipping Checklist
The Internal Data Logging Package includes:
•Interface cable
•Spreadsheet Interface Software (1 CD)
Both of these items are necessary for retrieving the stored data from the Picofluor™.
B.2 Hardware Requirements
•PC with Windows 95 or later
•MS Excel 5.0 or later
•At least 1 available serial port
B.3 Installation
1. Exit all Windows programs.
2. Insert the CD and run the setup program.
3. The setup wizard will install the necessary files. When the setup is complete, a SPREADSHEET
INTERFACE SOFTWARE icon will appear in the PROGRAMS menu.
4. Restart the computer.
B.4 Connecting
1. Using the provided cable, connect the 9-pin adapter of the cable into the available serial port of the
computer.
2. Plug the opposite end of the cable into the base of the Picofluor™.
3. Open the Spreadsheet Interface Software.
4. Click on the box to the right of the COM port icon to select the appropriate COM port. This is usually
COM port 1.
5. Click on the START button. The program will open MS Excel and should be ready for data transfer.
The boxes left of the words COM port and MS Excel should both be green.
6. Follow the directions from Section 5.2 for collecting and downloading data from the Picofluor™. Data
will automatically appear in the Excel spreadsheet. Be sure to save this data before closing the
software.
B.5 Troubleshooting
Difficulties can arise when parameters are set incorrectly or when cable connections are not tight enough.
Here are some common problems.
1. The box to the left of the COM port is red. This means that the COM port is not available. Causes:
•Another instrument or program (such as palm pilot/ hot sync) could be occupying the port, making
it unavailable. Make sure to close all programs of this type before downloading data
•The port selected is incorrect. Follow step 4 of connecting to choose another COM port.
2. All the lights are green, but no data is transferred, even though the instrument says “All data
downloaded.”
•The connection between the instrument and the computer is bad. Check and tighten the cable
connections. Make sure both ends of the cable are plugged in tightly.

Part Number: 998-0853
Version: 1.5
645 N. Mary Avenue
•
Sunnyvale, CA 94085
Phone: [phone] •Toll Free: [phone] •Fax: [phone]
www.turnerbiosystems.com.com