Isolated Perfused Organs

Overview:

The development of isolated perfused organ systems as a research technique has contributed significantly to medical research. Many animal organs can be isolated and studied in vitro. This is often done by perfusing the organ through its natural vascular system with an oxygenated physiological solution or whole blood using a peristaltic or syringe pump. The advantages of such a preparation include more reproducible results due to:

  • Isolation from circulating hormones
  • Free from sympathetic/parasympathetic influence
  • Unaffected by other organs

Organs that are commonly studied include the liver, kidney, heart, lungs, intestine, mesenteric arterial bed, and individual blood vessels. Rodent organs, in particular hearts and lungs, are the most commonly used isolated mammalian organs, but organs from larger animals such as pigs are also used and even organs from non-mammalian species such as fish.


 

Method:

For optimum use of the perfused organ system, it is crucial that the experimenter give careful thought to the design of the experiment, the donor organ, its maintenance and instrumentation. Isolated perfused organ systems require maintenance of the organ at physiological temperatures and perfusion with crystalloid solutions, such as Kreb’s or Tyrode’s solutions, or mixtures containing blood.

ADInstruments offer modular perfusion systems by Radnoti for isolated heart (Langendorff and Working Heart) studies. These systems can be used in constant pressure or constant flow, re-circulating or non-re-circulating experimental protocols.

ADInstruments have also developed the STH Pump Controller which can be used with a peristaltic pump such as the Minipuls 3 Peristaltic Pump to control both perfusate flow and pressure. They are both included with the compact “all-in-one” Langendorff System by Panlab. The Pump Controller allows the researcher to perfuse the organ at a set constant perfusate flow or, with the aid of a pressure transducer, set constant perfusate pressure.

ADInstruments also offer monitoring equipment such as amplifiers, electrodes, transducers and electrochemical sensors to measure physiological as well as biochemical parameters including:

  • Pressure: can be measured using a BP Transducer with a Bridge or BP Amplifier (single ended or differential; cmH2O to mmHg)
  • Flow rate: can be measured using a Flowmeter by Transonic Systems with a calibrated Tubing Flowsensor. Note: If the STH Pump Controller is used, a signal is provided which can be calibrated to determine the flow rate.
  • Temperature: can be measured with a T-type Thermocouple Probe and the T-type Pod.
  • Dissolved O2: oxygen levels in the perfusate line can be monitored with an Oxygen Electrode and suitable meter.
  • pH: can be measured with a small pH Electrode and the pH Amp.
  • Ions: can be measured with suitable ion selective and reference electrode. This includes ions such as sodium, potassium and calcium. The pH Amp is suitable as a meter and can be used with a separate reference electrode.
  • CO2 and NH3: can be measured with an appropriate electrode used with the pH Amp.
  • Cardiac Output (CO): can be measured using the Millar Pressure-Volume Catheters with the MPVS unit to directly measure end-diastolic volume (edv) and end-systolic volume (esv). Therefore, CO is simply: CO = [edv-esv] x HR

Related areas of interest include:

  • Langendorff Heart application page
  • Working Heart application page
  • Radnoti Tissue/Organ Bath Principles Technique Note: Information about various techniques and principles related to isolated tissue/organ research applications. Download (255 KB)
  • Glassware Maintenance and Post-Experimental Cleaning Technique Note: Information about maintaining and cleaning glassware and other components manufactured by Radnoti Technology Inc. Download (65 KB)

Software:

The LabChart Advantage:

(may require additional Modules and Extensions)

  • Units Conversion for easy calibration into appropriate units such as mmHg, °C, pH
  • Comments can be added in real time, viewed or moved at a later stage
  • The Dose Response Module allows for the calculation of dose response type data from LabChart recordings, automatically or manually, offline or online.
  • The Peak Analysis Module provides automatic detection and analysis of multiple, but not overlapping, signal waveforms from recordings, online or offline.
  • Fast data extraction, analysis and export (e.g. csv. or txt.) to other applications using Timed Add to Data Pad and Multiple Add to Data Pad
  • The LabChart GLP Client and GLP Server software offers a comprehensive audit trail, secure signing and a centralized user authorization system required for GLP and 21 CFR Part 11 (FDA) compliant environments.

LabChart 

LabChart software (for Windows and Macintosh) together with a PowerLab data acquisition system offers up to 32 channels of real-time data acquisition, data integrity, easy selection of hardware settings, powerful online and offline analysis, procedure automation, seamless extraction of experimental data and flexible display options. Additional acquisition and analysis functionality is provided with the use of specialized LabChart Extensions and LabChart Modules. Modules are available as part of LabChart Pro while Extensions are free for download from the website for existing LabChart users.

Dose Response Module

The Dose Response Module (Windows) provides easy analysis of dose response type data (response to stimulation by chemical, electrical or physical agonists) recorded in LabChart from various studies including:

  • Muscle contraction
  • Enzyme activity
  • Hormone secretion
  • Heart rate
  • Blood pressure
  • Membrane potential

This module provides:

  • Real-time or offline analysis
  • Automated or manual modes of analysis
  • Fast analysis of raw data to dose response parameters
    • Fast comment detection and conversion to dose response markers
    • Easy options for calculating different response parameters
    • Instant single or multiple dose response curves (Hill-curves) generation
    • Instant calculation of EC50 and Hill slopes
  • Export options to other software applications for further analysis

Peak Analysis Module

The Peak Analysis Module (Windows) provides automatic detection and analysis of multiple, but not overlapping, signal peaks in acquired waveforms.

It provides:

  • Real-time or offline analysis
  • Fast parameter extraction and analysis from single or multiple peaks
  • Automatic peak detection of an entire channel or a selection within a channel
  • Detection of positive or negative peaks
  • Provides several analysis presets suitable for general and specific types of waveforms
    • Evoked Response, Population Spike, Synaptic, Action Potential and Cardiac Action Potential
  • Instant tabulation of extracted parameters that can be exported to other applications
    • Height, width, area of the peak and time to peak

PV Loop Module

The PV Loop Module (Windows) records and analyzes left ventricular pressure and volume data for hemodynamic research in animals. Analyses can be conducted in real-time or offline.

It features:

  • Loop View- plots and displays the key cardiac parameters and allows individual loops to be reviewed and selected for inclusion/exclusion in analyses. Parameters include;
    • ESPVR (End Systolic Pressure-Volume Relationship). The slope of the ESPVR curve is an index of myocardial contractility; the steeper the ESPVR curve, the greater the cardiac contractility.
    • EDPVR (End Diastolic Pressure-Volume Relationship). The slope of the EDPVR curve is an indicator of ventricular compliance; the steeper the curve, the greater the ventricular stiffness.
  • Haemodynamics Table- reports parameters for each individual loop including:
    • Stroke Work
    • Cardiac Output
    • Ejection Fraction
    • Tau (the Isovolumic Relaxation Constant)
  • Analysis Plots
    • PRSW Plot: Displays Preload-Recruitable Stroke Work which is used as an index of contractility
    • dP/dt Max vs. EDV Plot: Shows the steepest slope during the upstroke of the pressure curve versus the end diastolic volume and is used as an index of contractility
    • PVA vs. EDV Plot: Shows pressure volume area versus the end diastolic volume which represents the mechanical energy generated by contraction of the ventricle
    • PVA vs. ESP Plot: Shows pressure volume area versus the end systolic pressure which represents the mechanical energy generated by contraction of the ventricle
    • Note: What the PVA vs. EDV, and PVA vs. ESP plots provide is a way of fitting a straight line to the data and obtaining an estimate of the slope parameter. This slope parameter can then be used a measure of contractility.
  • Automated Calibration
    • Cuvette calibration - for calculating absolute volume units using calibration cuvettes
    • Saline calibration - for determining parallel conductance of the ventricular wall (Vp)
       

GLP and 21 CFR Part 11

For those researchers working within a laboratory requiring GLP and 21 CFR Part 11 compliance the GLP Client and GLP Server are available for use with LabChart (Windows only) and PowerLab data acquisition systems. For more information, visit the Good Laboratory Practice application page or contact your nearest ADInstruments representative.

Hardware:

PowerLab Data Acquisition Systems

The PowerLab is a high-performance data acquisition unit capable of recording at speeds of up to 400,000 samples per second continuously to disk (aggregate). PowerLab units are compatible with instruments, signal conditioners and transducers supplied by ADInstruments, as well as many other third-party companies. In addition to standard single-ended BNC inputs, 4 differential Pod ports are also available for direct connection of Pod signal conditioners and appropriate transducers. Research PowerLab units include:

Research Systems

Compact “All-in-One” Langendorff Systems

Modular Langendorff and Working Heart Systems

Signal Conditioners

Pressure

Temperature

pH

  • ML165 pH Amp: Suitable for pH, temperature and ion-selective measurements in solution. Supplied with a RTD Temperature Probe.

Biopotentials

  • FE136 Animal Bio Amp: An isolated, high performance and software-controlled differential amplifier.
  • FE132 Bio Amp: A single channel, differential amplifier that is suitable for recording a range of biopotentials including Lead I, Lead II or Lead III ECG.
  • FE135 Dual Bio Amp: A dual channel, differential amplifier that is suitable for recording 2 independent biopotentials that share a common ground.
  • ML138 Octal Bio Amp: A differential amplifier that consists of eight electrically isolated differential input AC amplifiers.
  • GT201/F 16 Channel Bio Amp: A fully isolated, 16 channel stand-alone biological amplifier that consists of 8 modules, with each module containing two channel amplifiers that share a common ground input.

Instruments

Isolated Heart- Langendorff

Transit-Time Ultrasound Blood/Fluid Flowmeters

Millar Pressure Volume Systems (MPVS)

Transducers and Accessories

Pressure (fluid-filled) Transducers

Mikro Tip Pressure-Volume Catheters

Single Segment Pressure-Volume catheters (mice/rats)

  • PVR-1030 Mouse PV Catheter (1F, 4E, 3mm) - Non Repair
  • PVR-1035 Mouse PV Catheter (1F, 4E, 3.5mm) - Non Repair
  • PVR-1045 Mouse PV Catheter (1F, 4E, 4.5mm) - Non Repair
  • SPR-839 Mouse PV Catheter (1.4, 4E, 4.5mm)
  • SPR-847 Rat PV Catheter (1.4F, 4E, 9mm)
  • SPR-869 Rat PV Catheter (2F, 4E, 6mm)
  • SPR-838 Rat PV Catheter (2F, 4E, 9mm)
  • SPR-878 Rat PV Catheter (2F, 4E, 12mm)
  • SPR-858 Rat PV Catheter (2F, 4E, 14mm)
  • SPR-851 Rat PV Catheter (3F, 4E, 20mm)
  • SPR-853 Mouse Dual PV Catheter (1.4F, 2P, 4E, 4mm)
  • SPR-848 Mouse Dual PV Catheter (1.4F, 2P, 4E,4.5mm, Ap)
  • SPR-864 Mouse Dual PV Catheter (1.4F, 2P, 4E,4.5mm, Car)
  • SPR-901 Rat Dual PV Catheter (2F, 2P, 12mm, 4E)
  • SPR-902 Rat Dual PV Catheter (2F, 2P, 12mm, 4E 9mm Ap)

Multi Segment PV Catheters (dog/pig/sheep)

  • VENTRI-CATH-507 VC Catheter (5F, 12E, 7mm, DField, Pigtail, 122cm)
  • VENTRI-CATH-507S VC Catheter (5F, 12E, 7mm, DField, Straight, 122cm)
  • VENTRI-CATH-510 VC Catheter (5F, 12E, 10mm, DField, Pigtail, 122cm)
  • VENTRI-CATH-510S VC Catheter (5F, 12E, 10mm, DField, Straight, 122cm)
  • VENTRI-CATH-512 VC Catheter (5F, 12E, 12mm, DField, Pigtail, 122cm)
  • VENTRI-CATH-512S VC Catheter (5F, 12E, 12mm, DField, Straight, 122cm)
  • VENTRI-CATH-515 VC Catheter (5F, 12E, 15mm, DField, Pigtail, 122cm)
  • VENTRI-CATH-515S VC Catheter (5F, 12E, 15mm, DField, Straight, 122cm)
  • SPR-562-1 Multi-Seg PV Catheter (6F, 2P, 12E D-Field 7mm, 125cm)

Multi Segment PV Catheters (rabbits/cats)

  • SPR-877 Multi-Seg PV Catheter (3F, 10E D-Field 2.5mm, Straight, 120cm)
  • SPR-889 Multi-Seg PV Catheter (3F, 10E S-Field 3mm, U-tip, 80cm)
  • SPR-894 Multi-Seg PV Catheter (3F, 10E D-Field 4mm, U-tip, 80cm)

Thermocouple Temperature Probes

pH Sensors

  • MLA060 Redox Electrode: Suitable for operation from 0 to 60°C. The double junction is easily cleaned, making it suitable for biological samples.
  • MLA042 pH Electrode: Operates from 0 to 60°C and for a pH of 0 to 14. The double junction is easily cleaned and therefore the electrode is suitable for biological samples, in particular those of low ionic strength.
  • MI-405 Miniature Glass Electrode for pH: For use in general purpose analyses and in microchemical studies. Requires the use of an external reference electrode such as the MI-409 Miniature Reference Electrode.
  • MI-409 Miniature Reference Electrode: For use with any pH or ion selective electrodes and used with the ML165 pH Amp. Features an internal Ag-AgCl internal reference electrode with KCl filling solution.

Oxygen Electrodes

Biopotentials

Langendorff Accessories

Third-Party Electrodes
Third-party electrodes and meters may be connected to a PowerLab data acquisition system, provided that they have a ± 10 V analog output range. Such electrodes may include:

  • Ion Selective Electrodes and Microelectrodes
  • pH Field Effect Transistor (pHFET) Electrodes
  • Ion Selective Field Effect Transistor (ISFET) Electrodes
  • Fiber Optic Probes

 

Excessive thyroxine enhances susceptibility to apoptosis and decreases contractility of cardiomyocytes
Yun-Ying Wang, Bo Jiao, Wang-Gang Guo, Hong-Lei Che, Zhi-Bin Yu, Molecular and Cellular Endocrinology, 67-75, 2010

Mechano-Electric Feedback in the Fish Heart
S.M. Patrick, E White, H.A. Shiels, PLoS ONE, , 2010

Type 1 diabetic cardiomyopathy in the Akita (Ins2WT/C96Y) mouse model is characterized by lipotoxicity and diastolic dysfunction with preserved systolic function
Basu R, Oudit GY, Wang X, Zhang L, Ussher JR, Lopaschuk GD, Kassiri Z, American Journal of Physiology: Heart and Circulatory Physiology, H2096–H2108, 2009

Citations Database


The material on this page is provided in good faith and believed accurate at the time of writing. No responsibility will be taken, or liability accepted, for damages arising from the use of information herein. Readers are urged to check with respective manufacturers the accuracy of all product related information.

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