pyroscience FireStingO2 User manual

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FireStingO2
FIBER-OPTIC OXYGEN METER
USER MANUAL

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Document Version 3.07
Refers to Pyro Oxygen Logger Software version >3.1
The FireStingO2 is manufactured by
PyroScience GmbH
Hubertusstr. 35
52064 Aachen
Germany
Phone [phone]
Fax [phone]
Email [e-mail]
Internet www.pyro-science.com
Registered: Aachen HRB 17329, Germany

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TABLE OF CONTENT
1OVERVIEW ............................................................................. 6
2SAFETY GUIDELINES .............................................................. 8
3INTRODUCTION TO THE FIRESTINGO2...................................11
4SOFTWARE INSTALLATION...................................................13
5OXYGEN SENSOR TYPES.......................................................14
5.1 NEEDLE-TYPE SENSORS............................................................ 14
5.2 BARE FIBER SENSORS ................................................................17
5.3 ROBUST PROBES...................................................................... 19
5.4 SENSOR SPOTS........................................................................ 20
5.5 FLOW-THROUGH CELLS ............................................................ 22
5.6 RESPIRATION VIALS.................................................................. 23
5.7 CONNECTING THE SENSORS....................................................... 24
5.8 CLEANING AND MAINTENANCE OF THE SENSORS........................... 25
6THE SOFTWARE “PYRO OXYGEN LOGGER”............................ 27
6.1 MAIN WINDOW........................................................................ 27
6.2 SETTINGS................................................................................ 35
6.2.1 Basic Settings.....................................................................37
6.2.2 Advanced Settings............................................................. 38
6.2.3 Conditions in the Sample ................................................... 39
6.2.4 Temperature...................................................................... 41
6.2.5 Options ............................................................................. 43
6.3 OVERVIEW PANEL .................................................................... 45
6.4 RAW DATA WINDOW ................................................................46
7CALIBRATION WITH “PYRO OXYGEN LOGGER”......................48
7.1 CALIBRATION PROCEDURE.........................................................49
7.1.1 Calibration Mode: Factory .................................................. 51
7.1.2 Calibration Mode: 1-Point in Ambient Air ............................ 52
7.1.3 Calibration Mode: 1-Point in Water or Humid Air ................. 54
7.1.4 Calibration Mode: 2-Point in Ambient Air............................ 56
7.1.5 Calibration Mode: 2-Point in Water or Humid Air................. 58

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7.1.6 Calibration Mode: Custom Mode........................................ 61
7.2 CALIBRATION WITH A 1.GENERATION FIRESTINGO2....................... 65
7.2.1 Calibration Mode: Factory (1. Generation FireSting) ............ 66
7.2.2 Calibration Mode: 1-Point (1. Generation FireSting)............. 66
7.2.3 Calibration Mode: 2-Point (1. Generation FireSting) ............ 69
7.2.4 Calibration Mode: Custom (1. Generation FireSting) .............71
7.3 ADVANCED ADJUSTMENTS......................................................... 72
8CALIBRATION STANDARDS...................................................74
8.1 THE AIR CALIBRATION STANDARD .............................................. 74
8.1.1 Ambient Air ....................................................................... 76
8.1.2 Water-Vapor Saturated Air .................................................77
8.1.3 Air Saturated Water............................................................77
8.2 THE 0% STANDARD.................................................................. 78
8.2.1 Water Mixed with a Strong Reductant ................................ 78
8.2.2 Water Flushed with Nitrogen Gas....................................... 79
8.2.3 Nitrogen Gas ..................................................................... 79
9CALIBRATION OF CONTACTLESS SENSORS ..........................80
9.1 CALIBRATION PROCEDURE......................................................... 80
9.2MANUAL BACKGROUND COMPENSATION .................................... 81
10 TEMPERATURE MEASUREMENT............................................83
10.1 AVAILABLE EXTERNAL TEMPERATURE SENSORS............................ 83
10.2 INTERNAL TEMPERATURE SENSOR .............................................. 83
10.3 AUTOMATIC TEMPERATURE COMPENSATION................................ 84
11 ANALOG OUTPUT AND AUTO-MODE ....................................86
11.1 ANALOG OUTPUT ....................................................................86
11.2 AUTO-MODE...........................................................................89
11.3 ADVANCED AUTO-MODE .......................................................... 91
12 APPENDIX.............................................................................93
12.1 SPECIFICATIONS OF THE FIRESTINGO2......................................... 93
12.2 EXTENSION PORT X1 ................................................................ 95
12.2.1 Connector X1 (Power, Digital Interface, Analog In).......... 95
12.2.2 Connector X2 (Analog Output) ..................................... 100
12.3 TROUBLESHOOTING ................................................................101

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12.4 MEASURING PRINCIPLE ........................................................... 102
12.5 OPERATING SEVERAL FIRESTINGO2 IN PARALLEL......................... 104
12.6 DEFINITION OF OXYGEN UNITS................................................. 105
12.7 TABLE OF OXYGEN SOLUBILITY .................................................107
12.8 EXPLANATION OF THE SENSOR CODE ........................................ 109

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1Overview
The compact USB-powered fiber-optic oxygen meter FireStingO2
with 1, 2, or 4 channels unifies several innovative technological
improvements making it the new standard of high precision
oxygen sensing with fiber-optical oxygen sensors (optodes). The
FireStingO2 utilizes a measuring principle based on
red light excitation and lifetime detection in the near infrared using
unique luminescent oxygen indicators (REDFLASH technology, see
Appendix 12.4 for more details). It is a multipurpose oxygen meter
•enabling measurements in water as well as in gas phases,
•working with fiber-based sensors comprising several size
classes like microsensors, minisensors, or robust probes,
•working with contactless sensors like sensor spots, flow-
through cells or respiration vials, and
•working with full range and trace oxygen sensors.
The FireStingO2 has integrated atmospheric pressure and
humidity sensors for a precise and easy sensor calibration, but also
for automatic pressure compensation of the oxygen
measurements. Furthermore, the FireStingO2 offers 4 analog
outputs and a built-in temperature port for an external
temperature sensor allowing automatic temperature
compensation of the oxygen measurement. If independent
temperature compensation for all oxygen channels is needed, a
USB-powered temperature extension module TeX4 can be easily
fixed below the FireStingO2 meter with a smart docking
mechanism. The user-friendly Pyro Oxygen Logger software
allows operation of several FireStingO2 meters in parallel as a
multi-channel system.
Along with the FireStingO2 we offer turnkey motorized
microprofiling setups for measurements of depth-profiles of
oxygen concentration in semi-solid environmental samples and

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along micro-gradients at high temporal and spatial resolution, as
well as customized OEM solutions.
More information concerning our products can be found at
www.pyro-science.com
or contact us under [e-mail].
We would be pleased to serve you concerning all needs for high
precision oxygen sensing.
Your PyroScience Team

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2Safety Guidelines
The FireStingO2 is a laboratory instrument to be used with fiber-
optic oxygen sensors (optodes) from PyroScience for measuring
oxygen at high precision and resolution. In order to guarantee an
optimal performance of the FireStingO2 please follow these
operation instructions and safety guidelines.
If any problems or damage evolve, please disconnect the
instrument immediately, mark it to prevent any further use and
consult PyroScience for repair or maintenance service. The
FireStingO2 should not be manipulated or opened by unauthorized
persons, only by PyroScience or persons advised directly from
PyroScience.
Please note that opening the housing will void the warranty. There
are no serviceable parts inside the device.
The FireStingO2 and sensors should be kept and stored outside the
reach of children in a secure place under dry and clean conditions
at room temperature, avoiding moisture, dust, corrosive
conditions and heating of the instrument. This device and the
sensors are not intended for medical, military or other safety
relevant areas. They must not be used for applications in humans;
not for in vivo examination on humans, not for human-diagnostic
or therapeutic purposes. The sensors should not be brought in
direct contact with foods intended for consumption by humans.
The FireStingO2 should be used in the laboratory by qualified
personal only, following the operation instructions and safety
guidelines of this manual. Please follow the appropriate laws and
guidelines for safety like EEC directives for protective labor
legislation, national protective labor legislation, safety regulations
for accident prevention and safety data-sheets from manufacturer
of chemicals used during measurements.

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Calibration and application of the sensors, data acquisition, data
processing and data publication is on the user's authority.
When used in the field, the environmental conditions (like high
humidity, dust, exposure to direct solar radiation) may cause
damage or interference of the FireStingO2, which is on the user's
authority.

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Before using the FireStingO2 and its sensors, read carefully the
instructions and user manual for the oxygen meter FireStingO2.
In case of problems or damage, disconnect the instrument and
mark it to prevent any further use! Consult PyroScience for
advice! There are no serviceable parts inside the device. Please
note that opening the housing will void the warranty!
The FireStingO2 is not watertight, is sensitive to corrosive
conditions and to changes in temperature causing
condensation. Avoid any condition (e.g. direct sun light) causing
a heating of the device above 50°C (122°F) or below 0°C. Avoid
any elevated humidity causing condensing conditions.
Handle the sensors with care especially after removal of the
protective cap! Prevent mechanical stress to the fragile sensing
tip! Avoid strong bending of the fiber cable! Prevent injuries
with needle-type sensors!
Calibration and application of the sensors is on the user’s
authority, as well as data acquisition, treatment and
publication!
The sensors and the oxygen meter FireStingO2 are not intended
for medical, diagnostic, therapeutic, or military purposes or any
other safety-critical applications. The sensors must not be used
for applications in humans and should not be brought in direct
contact with foods intended for consumption by humans.
The FireStingO2 and sensors should be used in the laboratory
only by qualified personal following the user instructions and
the safety guidelines of the manual, as well as the appropriate
laws and guidelines for safety in the laboratory!
Keep the sensors and the oxygen meter FireStingO2 outside the
reach of children!

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3Introduction to the FireStingO2
The FireStingO2 is an optical oxygen meter that is compatible with
a broad range of oxygen sensors from PyroScience:
microsensors and minisensors (retractable needle-type,
fixed needle-type, or bare fiber),
robust probes and
contactless sensors (sensor spots, respiration vials, flow-
through cells)
Most sensors are available in versions for the full range (0-100%
O2) and for the trace range (0-21% O2). The optical detection
technology is based on the unique oxygen-sensitive REDFLASH
indicators which use red light excitation and lifetime detection in
the near infrared (see Appendix 12.4 for more details).
The FireStingO2 is a high precision, compact USB-powered fiber-
optic oxygen meter with 1, 2 or 4 channels for measurements in
the laboratory. Additionally, one temperature sensor can be
connected for automatic temperature compensation of the oxygen
measurement. The integrated sensors for atmospheric pressure
(mbar) and relative humidity (%RH) of the ambient air enable a
precise and easy sensor calibration, as well as automatic pressure
compensation of the oxygen measurements.
The FireStingO2 is operated via a Micro-USB connection to a
PC/tablet with a Windows operation system. The included logging
software Pyro Oxygen Logger provides comfortable calibration
and logging functionality.
The FireStingO2 is a laboratory instrument, and if used in the field,
please protect the FireStingO2 from heating, moisture and
corrosion.

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The FireStingO2 comes with 1, 2, or 4 channels (connectors 1to 4)
on the right side panel for up to 4 fiber-optic oxygen sensors and
one connector (T) for a temperature sensor.
The Micro-USB connector on the left side panel provides the
energy supply and the data exchange with the PC. Right-hand side
of it, a connector X1 for power and digital interface (7-pins) and a
connector X2 for analog output (5 pins) is located. The holes
function as air inlets for the internal temperature, pressure and
humidity sensors. Please avoid covering these holes to ensure free
air circulation towards the internal sensors.

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4Software Installation
IMPORTANT: Do not connect the FireStingO2 to your PC before
the Pyro Oxygen Logger software has been installed. The software
will install automatically the appropriate USB-drivers.
System requirements:
PC with Windows XP/Vista/7/8 (but not Windows RT) and
min. 200 MB free disk space
Installation steps:
•download the installer package for the newest version of
the Pyro Oxygen Logger software from the PyroScience
homepage: www.pyro-science.com/downloads.html
•unzip and start the installer and follow the instructions
•connect the FireStingO2 with the Micro-USB cable to the
computer. The red logo will flash shortly indicating the
correct startup of the oxygen meter.
After the successful installation a new program group „Pyro
Oxygen Logger“ is added to the start menu, and a short-cut named
"Oxygen Logger" can be found on the desktop.

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5Oxygen Sensor Types
For an overview of all available oxygen sensor types, please see
also: http://www.pyro-science.com/overview-fiber-optic-oxygen-
sensors.html.
5.1 Needle-Type Sensors
The needle-type oxygen sensors are composed of a fiber-optical
cable (C) with an ST-plug (P) for connection to the FireStingO2
meter, a metal housing (H, fixed or retractable), and a syringe
needle (N; 40 mm standard length) including the fiber with a
fragile sensing tip and protected with a protective cap (PC). On
each sensor, a specific Sensor Code (SC) is attached to the cable.
The oxygen-sensitive REDFLASH indicator is immobilized at the
tip of the sensor, which can be retractable (R)or fixed (F) in the
housing.
The needle-type oxygen sensors are offered with different tip
diameters, including retractable microsensors with a tapered

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sensor tip of ca. 40-60 µm in diameter (A) and minisensors with a
flat sensor tip of 230 µm or 430 µm (B), as well as fixed minisensors
with 0.5 mm and 1.1 mm tip diameter (C).
The retractable sensors are available as microsensors (OXR50; A)
and as minisensors (OXR230, OXR430; B). The retractable sensor
tip allows sensor insertion
through septa or package
material, and subsequent
extension of the sensor tip into
the sample for oxygen measurements. The sensors are shipped in

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position 0 (retracted position) of the push button (PB) to protect
the tip from breaking.
The protective cap (PC) covering the needle has to be removed
carefully before using the sensor.
CAUTION: Remove only the protective cap, but not the needle!
Then the sensor should be fixed in a stable set-up like a solid
laboratory stand or a micromanipulator mounted on a heavy stand
from PyroScience. Move gently the push-button from the
retracted position 0to one of the extended positions 1-3. In
position 1 the sensor tip is flush with the needle tip, whereby in
positions 2 or 3 the fragile sensor tip extends ca. 6 mm or 12 mm
out of the needle.
Please take care when handling the needle-type sensors to prevent
injuries and breaking of the sensor. Ensure enough space in front of
the fragile sensor tip, especially when pushing it out!
The needle-type microsensors (OXR50; A) are appropriate for high
resolution measurements in semi-solid samples, like sediments,
biofilms or soils with PyroScience turnkey motorized micro-
profiling setups. They allow measurements in small sample
volumes, along steep gradients and of depth-resolved oxygen
profiles at high spatial resolution.

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Please always use a stable stand and a micromanipulator to move
the sensor into a semisolid sample like sediment, biofilm or soil.
The minisensors are standard sensors for robust long-term oxygen
measurements in gaseous or aquatic environments. Fixed
minisensors with a mechanically protected tip (OXF500PT; C) can
be used for insertion through a rubber septum without addition of
air to gas samples. Here the very sensor tip is flush with the tip of a
beveled needle and glued into it.
In case of fixed minisensors with a tip diameter of 1.1 mm
(OXF1100; C), the fixed sensor tip extends 1 mm from the needle
tip and is hence unprotected after removal of the protective cap.
Avoid any contact to harsh surfaces!
The needle-type oxygen sensors are also optionally available with
optical isolation for measurements in samples with auto-
fluorescence. With this optical isolation, interferences between the
REDFLASH indicator luminescence and autofluorescence, e.g.
from photosynthetic pigments, are reduced. However, the optical
isolation causes an increase in the response time t90, which is the
time for 90% of the total sensor signal change (see respective
Sensor Specifications).
5.2 Bare Fiber Sensors
The bare fiber sensors are identical
to the retractable needle-type
oxygen sensors, except that they do
not contain a housing. They
comprise only the bare optical fiber
with the REDFLASH indicator
immobilized on the sensor tip,
secured during transportation and storage in a protective plastic
tube (PT). The fiber-optical cable (C) is connected with the ST-plug

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(P) to the oxygen meter FireStingO2. On each sensor, a label (L)
with a specific Sensor Code is attached to the fiber-optic cable.
The protective plastic tube (PT) covering the bare fiber has to be
removed carefully before using the sensor. For this, fix the plastic
tube in a stable laboratory stand, loosen the clamping screw and
carefully pull out the bare fiber sensor.
NOTE: After removal of the protective plastic tube the fragile
sensor tip is unprotected and sensitive to breakage!
CAUTION during handling to avoid breakage or injuries!
The bare fiber sensors
are available both as
microsensors (OXB50;
A) and as minisensors
(OXB230, B; OXB430,
C). They can be used
with customized
housings with
complex geometries
or they might be
inserted directly into samples ("implantable sensors").

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5.3 Robust Probes
The Robust Oxygen Probes are
composed of a black fiber and a
stainless steel tubing 3 mm in
diameter, and 30 or 100 mm in
length (e.g. item no. OXROB3,
OXROB10). The oxygen
sensitive part is the disc 3mm in
diameter located at the very end of the tubing (green disc in image
below).
The Robust Oxygen Probes can be optionally obtained with an
optical isolation (black coating) on the sensing surface (item no.
OXROB3-OI, OXROB10-OI), which is recommended for
applications with strong external illumination like direct sun light.
The Robust Oxygen Probes can be used for long-term
measurements in gases and liquids. The whole sensor including its
cabling is completely submersible in water and specified for
longtime submersion in liquid media.
In contrast to the needle-type sensors, the sensitive sensor tip of
the robust probe is protected by the steel tubing and is more
robust for standard laboratory applications. Due to the
significantly bigger dimensions of the robust probes compared to
our micro- and minisensors, diffusion limitation in liquid samples of

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oxygen towards the 3 mm tip has a measurable effect on the
response time. For optimum operation conditions it is therefore
recommended to utilize these robust probes in permanently stirred
liquids (this diffusion limitation is negligible in gaseous samples;
see also the respective sensor specifications).
Ensure that the complete sensing surface of the Robust Oxygen
Probe is covered completely by the sample and prevent
mechanical stress (e.g. scratching) to the sensing surface!
5.4 Sensor Spots
PyroScience offers a
range of contactless
oxygen sensors,
comprising sensor
spots, respiration
vials and flow-
through cells. The
sensor spots are
coated with the
REDFLASH indicator
with PET foil or glass
as carrier material.
The sensor spots allow oxygen measurements in closed vessels in
combination with the FireStingO2 equipped with Optical Fibers
(e.g. item no. SPFIB or SPFIB-CL2). The sensor spots have
standard diameters of 5 mm (item no. OXSP5) and can be
optionally obtained with an optical isolation (black coating) on the
sensing surface (e.g. item no. OXSP5OI,OXSG8OI). Optical
isolation is recommended if strong illumination
might disturb the measurement or if no light shall
be introduced into the sample.