fluoroplex - kys t

www.ionoptix.com

IonOptix LLC 309 Hillside Street, Milton, MA 02186, USA T: 1‐617 696‐7335 F: 1‐617 698‐3553 E: [email protected] IonOptix Ltd 1a Dartmouth Ter., Ranelagh, Dublin 6 , IRL T: +353 1 685 4800 F: +353 1 443 0784 W: www.ionoptix.com www.ionoptix.com

USA T: 1‐617 696‐7335 F: 1‐617 698‐3553 E: [email protected] Europe T: +353 1 685 4800 F: +353 1 443 0784 W: www.ionoptix.com IonOptix

FluoroPlex Tissue Bath Fluorometry

Complete, turnkey system for high-speed fluorescence recording

• 250 ratiometric calcium measurements/second

• 1000 single wavelength measurements/second

Table of Contents

System Overview ....................................................................... 2

FluoroPlex Illumination ............................................................... 3

FluoroPlex MultiPlexer & Detection ............................................... 4

FluoroPlex Controller .................................................................. 5

ADI PowerLab and LabChart ........................................................ 6

Radnoti Tissue Baths and Force Detection ..................................... 7

Example Data ......................................................................... 8-9

IonOptix Systems .................................................................... 10

Contact ...................................................................... Back Cover

June 2012

For many years, a vast number of tissue and whole organ experiments have been done in water-jacketed tissue chambers. These experiments can be as simple as the measure-ment of force generated by smooth or striated muscle tissue or as elaborate as monitor-ing the EKG and chamber pressures from a beating heart. Multiple preparations are often examined simultaneously and it is not uncommon to arrange arrays of 4 or even 8 tissue chambers to increase data throughput. With the introduction of the IonOptix FluoroPlex, it is now possible to add fluorescence ion recording to tissue bath experi-ments in parallel with force, pressure, or other measurements. The ability to monitor regulatory intracellular ion levels promises to advance the insights revealed by this work-horse methodology.

The FluoroPlex epifluorescence illumination and detection assembly lies at the heart of the complete FluoroPlex Tissue Bath Fluorometry System. The system includes not only the FluoroPlex but also a suite of force transducer/ amplifier combinations and spe-cially-designed tissue baths built by Radnoti Glass Technologies Inc. to accommodate the solution-resistant tip of our liquid light guides. The complete system also includes an optional light-tight enclosure to house the Radnoti baths and stand, as well as an optional Radnoti thermo circulator. Data is handled by the optional ADInstruments PowerLab interface and LabChart and Scope software. The FluoroPlex System allows fast fluores-cence recordings from multiple tissue baths with simultaneous force measurements. Sampling occurs via two modes: burst and continuous. Burst mode sampling occurs at 1000 Hz with single wavelength excitation and 250 Hz with dual excitation. Continuous mode ratiometric sampling occurs at up to 10 Hz for four baths. Sampling modes, fre-quencies, duration and associated tasks such as signal averaging and background collec-tion are programmed within a simple user interface.

FluoroPlex System Overview

Light Paths: -- Path 1 -- Path 2 -- Path 1 or 2 – Emission

Excitation Source (top)

Bath Multiplexer

To Tissue Baths 5-8

Tissue Baths 1-4

Emission Filter Block (side)Sensor

The FluoroPlex epifluorescence illumination and detec-tion assembly is built for fast quantitative fluores-cence photometry collected from multiple tissue baths. Tissue fluorometry is completely automated through a stand-alone controller and is capable of both single and dual excitation modes. The FluoroPlex LED illumination system is designed to optimize speed and light throughput. With its intelligent controller, ingenious light path, and fast wavelength switching, the FluoroPlex promises to provide critical new in-sights that advance your research.

Emission Filter Block (side)


FLUOROPLEX LIGHT PATH

FluoroPlex Illumination

FLUOROPLEX

High Speed Dual Excitation Light Source

Features • LED illumination source: monochromatic

illumination with UV and Vis options.

• Excitation Intensity Control: Each LED can be independently controlled via gradu-ated, locking 10-turn potentiometers. The pots allow fine, calibrated control over illu-mination intensity.

• Electronic wavelength switching: ideal for dual excitation, permits rapid wavelength switching for quantitative ratiometric dyes such as the calcium probe Fura-2.

• Liquid light guides: providing direct inter-rogation of isolated tissue, liquid light guides are a highly efficient and flexible means of delivering and collecting light. Systems can be equipped with up to eight light guides to maximize experimental throughput and resources. The light guides are solution resistant, allowing them to be positioned within the baths in close proximity to the tissue.

• Shuttering. The FluoroPlex utilizes elec-tronic shuttering between sampling periods to minimize light exposure, thereby avoiding dye bleaching and photo-damage to the preparation.

• Innovative light path: complete with wavelength filtering specific to your fluores-cent probe, the light path was painstakingly designed to minimize background while preserving signal.

• High-Speed: Burst mode sampling rate is 1000 Hz (250 Hz dual excitation); Continu-ous mode sampling rate for four baths is 10Hz.

The MultiPlexer delivers fluorescence illumination light to as many as eight light guides. The MultiPlexer consists of a focusing lens, microstepping motor-mounted mirror and light guide housing assembly. The MultiPlexer can be driven in two modes to increase temporal resolution and sampling rates or to increase data throughput from multiple baths. Focused excitation light is steered into one of up to eight available light guides. Excitation light emerging from the light guide directly interrogates the sample.

Fluorescence emission is then collected through the same light guide and synchronously reflected by the MultiPlexer mirror back into the FluoroPlex. The emis-sion light passes through a vertically reflecting dichroic and is reflected through an emission filter before reach-ing a photodiode with user controlled gain and offset, enabling optimization of the photodiode’s sensing effi-ciency. The photodiode’s output is sent to and processed by the FluoroPlex Controller.

FluoroPlex MultiPlexer & Detection

Fast, High-Throughput Recording

Features • Microstepping MultiPlexer: offering pre-

cise control of light guide selection, the multiplexer steers light to as many as eight light guides.

• Fast MultiPlexing: Switching to adjacent light guides (moving from one bath to an-other) requires less than 1/40th of a sec.

• Optics: Emission filtering and dichroics are optimized for several fluorophores, including the quantitative ratiometric calcium indicator Fura-2.

• Photodiode Sensor: Ideally suited to the fluorescence of whole tissue, the FluoroPlex offers a large linear dynamic range to collect from low resting calcium levels to peak calcium.

• Sensor Control: The photodiode can be optimized through graduated, locking 10-turn potentiometers. The pots allow fine, calibrated control over both signal amplifica-tion and offset.

MULTIPLEXER

MULTIPLEXER & EMISSION SCHEMATIC

Emission Filter Block (side)


The FluoroPlex Controller provides a convenient digital interface and an intuitive experiment programming dialog to facilitate data acquisition. FPX hardware func-tions can be easily controlled through its intelligent native software.

The controller interface offers the ability to select be-tween sampling modes, collect background and specify sampling rates and signal averaging parameters. De-signed to work in conjunction with existing laboratory interfaces and software, the controller houses a full suite of analog data outputs as well as a digital output for easy synchronization to external events.

FluoroPlex Controller

System Integration & Control

Features

• Stand-alone Controller: completely auto-mated, intuitive interface allows easy selec-tion of the illumination protocol.

• Selectable Sampling Modes: Protocols can be written in "Burst mode" for high-speed interrogation of an individual specimen before changing to the next channel, or "continuous mode" for continuous sampling of all baths.

• Background Subtraction: Background values are easy to collect and store. These values are automatically subtracted before data output. For dual excitation dyes, val-ues are automatically ratio’ed before data output.

• Selectable Sampling Rates: User-defined, variable sampling frequency enables investi-gators to choose the best rates to resolve events while minimizing photobleaching and phototoxicity.

• Signal Averaging: The FluoroPlex allows signal averaging for noise reduction.

Outputs

• Data Out: Eight channels of 12-bit analog data outputs for either eight background-subtracted ratiometric values, eight back-ground subtracted single emission values, or four raw numerator/four raw denominator values.

• Raw Out: Two channels of continuous, raw 12-bit analog values.

• Digital Out: TTL pulse output allows syn-chronization with external hardware, which can be coordinated with one of four hard-ware events: (1) at the beginning of an experiment protocol, (2) at the beginning of an acquisition cycle when the MultiPlexer is positioned at the first bath, (3) accompany-ing each move to a new bank, or (4) accom-panying the output of each new data point.

FPX CONTROLLER

We offer AD Instruments PowerLab data acquisition package, including LabChart software for complete data acquisition and analysis systems. As the most recog-nized name in data acquisition for over two decades, AD Instruments products have been installed in thousands of research labs around the world. The PowerLab inter-face and LabChart software offer an ideal solution to digitize and analyze the outputs from our FluoroPlex quantitative fluorometry system.

ADI’s PowerLab systems offer versatile data acquisition and analysis solutions for life science research applica-tions. The systems are used to record and analyze physiological signals from human and animal subjects. In one compact unit, PowerLab systems perform the func-tions of chart recorders, XYT plotters, digital voltmeters and storage oscilloscopes.

PowerLab data acquisition units are smart peripheral devices that perform data acquisition, signal conditioning and pre-processing.

LABCHART

ADInstruments PowerLab & LabChart

Acquisition/Digitization of Quantitative Fluorescence Data

PowerLab Features

• Compact, all-in-one unit with small bench footprint

• External signals are acquired through analog inputs and amplified before being digitized

• The signal is multiplexed to an analog-digital converter

• The digitized signal is transmitted to the computer using USB connection

Lab Chart Features

• Continuously record and display up to 32 channels of data (up to 16 raw and up to 32 calculated signals)

• Perform online or offline analysis

• Display a range of specialized View windows including Scope View, Zoom View and XY View

• Automatically recognize PowerLab models, amplifiers and smart transducers

• Automatically export recorded values to other graphical or statistical packages

Over four decades the Radnoti brand has become syn-onymous with high quality at a reasonable price. Radnoti was founded as a glass blowing company but over the years it has evolved into a full service manufacturer of research systems used in a range of life science areas. Radnoti manufactures and supplies a complete range of glassware as well as associated hardware and software.

IonOptix worked with Radnoti to develop tissue baths that would accept and position the solution-resistant FluoroPlex liquid guides for direct interrogation of tissue fluorescence. We also co-developed a light-tight enclo-sure to fit the standard Radnoti 4-bath system, allowing researchers to perform light-sensitive fluorescence measurements on the bench in a well lit environment.

Radnoti Tissue Baths & Force Detection

Multi-Bath Tissue Systems

Features

• Radnoti Tissue Bath Systems are recom-mended for passive and stimulated muscle preparations for cardiac muscle, skeletal muscle, smooth muscle such as aortic rings, bladder or intestine as well as many other experiments.

• 5ml, 10ml, 25ml and 50ml baths available as standard selections with the system. Specialty tissue baths may be requested at an additional charge. The purchase of sev-eral different size baths for the unit is also an option.

• Extremely precise independent isometric force transducers with two force range settings (0-2gm, 0-20gm), based on pico-farad capacitance converted to DC voltage.

• Superb temperature control due to our water-jacketed tissue bath and components. Start-up to 37°C in 15 mins with less than 0.01° C error.

• Quick and easy disassembly due to our quick-disconnect tubing.

• Easy system maintenance and long-term durability due to GLP-compliant open archi-tecture and sturdy steel base that is coated to resist corrosive salt solutions..

• Easy cleaning: our crystal clear borosilicate glassware is fully autoclavable and unlike plastic systems, are not affected by agents and proteins.

• Easy replacement: Individual components allow for easy replacement in the event of breakage and can be replaced without fac-tory return.

• Customized configurations available. As with all Radnoti systems, the modular design allows for extreme flexibility and system reconfigurations.

• Optional additional reservoirs may be pur-chased to perform complex drug additions and wash out protocols.

• Available as a complete system, with Iso-metric Force Transducers and amplifiers, and Thermal Circulator Water Bath

TISSUE BATH SYSTEM

Example Data Sectioned Heart

0.0 0.5 1.01.0

1.5

2.0

Fluo

resc

ence

(a.u

.)

Time (s)

Control 100 nM ISO

60 65 70 305 310

1.5

2.0

2.5

3.0

3.5

4.0

Con

trac

tile

forc

e

Time (s)0.0 0.5 1.0

1.5

2.0

2.5

3.0

3.5

4.0

Cotra

ctile

forc

e

Time (s)

Control 100 nM ISO

65 70 305 310

1.0

1.2

1.4

1.6

1.8

2.0

Fluo

resc

ence

(a.u

.)

Time (s)

Ventricular Papillary Muscle (± Iso)

Fura Ratio

Fura Ratio

Left Atrium (± Iso)

Fura Ratio

Fura Ratio

Example Data (Continued)

Skeletal Muscle

Epitrochlearis Muscle

Single Stimulation,

0.5 millisec pulse

Fluo

Force

Bath 1 Fura

Bath 1 Force

Bath 2 Fura

Bath 2 Force

50 Hz tetanic

stimulation for

0.5 sec

Time (seconds)

Time (seconds) Data coutesty of Anthony Payne and Tom Clanton (University of Florida)

Force (grams)

Force (grams)

Fura Ratio

Fluo

rescen

ce (cou

nts)

Diaphragm Muscle

1Hz 10Hz 20Hz

VESSEL DIAMETER SYSTEM

IonOptix Systems

Our goal is to support the scientific research community with an array of systems that meet experimental demands while adhering to our philosophy of developing innovative high-performance products at fair prices. The following are some of our application-driven systems currently available. Visit us at www.IonOptix.com for more information.

Calcium and Contractility/ Diameter

• Myocyte Calcium and Contractility Recording Systems

• HyperSwitch Configuration

• µStep Configuration

• Vessel Calcium and Diameter Recording System

• Myocyte Contractility System

• Vessel Diameter System

Tissue Bath Fluorometry

• FluoroPlex

General Photometry

• Fluorescence Photometry System

Cell Pacing

• C-Pace EP Cell Culture Pacing

Myocyte Harvesting

• Myocyte Harvesting System

CELL CULTURE PACING

MYOCYTE CALCIUM AND CONTRACTILITY SYSTEM

www.ionoptix.com IonOptix LLC 309 Hillside Street, Milton, MA 02186, USA T: 1‐617 696‐7335 F: 1‐617 698‐3553 E: [email protected] IonOptix Ltd 1a Dartmouth Ter., Ranelagh, Dublin 6 , IRL T: +353 1 685 4800 F: +353 1 443 0784 W: www.ionoptix.com www.ionoptix.com

USA T: 1‐617 696‐7335 F: 1‐617 698‐3553 E: [email protected] Europe T: +353 1 685 4800 F: +353 1 443 0784 W: www.ionoptix.com IonOptix

Company History IonOptix makes quality ratiometric fluorescence and cell dimensioning data acquisition systems. We have been making reasonably priced, high performance systems since 1990.

IonOptix prides itself on post-sale customer support. Telephone and email support is available on an unlimited basis. More importantly, every system sale includes a one to two day installation visit to set up the system and train the customer. We consider this training to be critical as it gets the customer up and running as quickly as possible. We run experiments with your preparations during the visit to be assured that all technical issues particular to the experimenter’s preparations have been covered.

“The Barn” — IonOptix Headquarters in Milton, Massachusetts

Resellers Our good friends at Cairn Research are currently our only resellers. They sell our components in conjunction with their fluorescence and electrophysiology systems.

Cairn Research

www.cairn-research.co.uk

Authorized Representatives In efforts to afford this high level of service to all our customers, IonOptix has entered into agreements with local representatives in several countries. The represen-tatives have been chosen based on their technical and biological expertise, familiarity with our products, and on their customer service skills.

Bioprobes Ltd. China, Hong Kong www.bioprobeshk.com www.bioprobeschina.com

Primetech Japan www.primetech.co.jp

Scitech Korea Inc. Korea www.scitechkorea.co.kr

Kuo Yang Scientific Corp. Taiwan, Republic of China www.kuoyang.com.tw

SDR Clinical Technology Australia, New Zealand www.sdr.com.au

Commat Ltd. Turkey www.commat.com.tr

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