Validation of the use of field effect transistors for extracellular signal recording in pharmacological bioassays.
The use of field effect transistors (FETs) in biomedical research has been in rapid progression in recent years. The present study aims to demonstrate a quantitative use of these devices in pharmacological bioassays. FETs were made as a 4 x 4 matrix of gates with a width of 200 microm separating each gate. The surface of the FETs (silicon oxide), covered with a layer of laminin, fibronectin, or nitro cellulose was suitable for cell adhesion. The cultured dissociated cardiac myocytes were spontaneously active within 24 to 48 h after initial plating. Simultaneous intracellular patch clamp recordings were used to verify the electrophysiological signals of cells that were coupled to the gates. All positive chronotropes (isoproterenol, norepinephrine) and negative chronotropes (verapamil, carbamylcholine, SDZ PCO400) showed their characteristic effects on heart cells in terms of changes of beat frequency. As the myocytes were in a complete syncitium on each FET, the cells need not be directly coupled to the gate in order to detect any ionic changes. This enables global cellular responses to be analyzed. The system also offers an opportunity to study the interconnections and communications between different cells. Furthermore, the changes of signal shapes in the presence of different agents could also be detected. The present study demonstrates how versatile and sensitive this recording system is in distinguishing different ionic signal shapes. The authors believe that this system has the potential to replace some currently employed in vitro methods, offering an alternative, which can substantially reduce animal use in pharmacological experiments.
Yeung
C K
CK
Max-Planck-Institute for Polymer Research, Ackermannweg 10, Mainz, D-55128, Germany. ckyeung@dera.gov.uk
Ingebrandt
S
S
Krause
M
M
Offenhäusser
A
A
Knoll
W
W
D000200:Action Potentials; D000818:Animals; D001578:Benzopyrans / Q000494:pharmacology; D001681:Biological Assay / Q000295:instrumentation* / Q000379:methods; D015374:Biosensing Techniques / Q000295:instrumentation* / Q000379:methods; D002217:Carbachol / Q000494:pharmacology; D002317:Cardiovascular Agents / Q000494:pharmacology; D002478:Cells, Cultured; D003517:Cyclopentanes / Q000494:pharmacology; D004305:Dose-Response Relationship, Drug; D004563:Electrochemistry / Q000295:instrumentation; D004566:Electrodes; D006321:Heart / Q000187:drug effects / Q000196:embryology / Q000502:physiology; D007545:Isoproterenol / Q000494:pharmacology; D009206:Myocardium / Q000166:cytology; D009638:Norepinephrine / Q000494:pharmacology; D018408:Patch-Clamp Techniques; D051381:Rats; D017207:Rats, Sprague-Dawley; D012666:Semiconductors; D014173:Transistors, Electronic; D014700:Verapamil / Q000494:pharmacology
D001578:Benzopyrans; D002317:Cardiovascular Agents; D003517:Cyclopentanes; C073091:SDZ PCO 400; D002217:Carbachol; D014700:Verapamil; D007545:Isoproterenol; D009638:Norepinephrine