A chamber to permit invasive manipulation of adherent cells in laminar flow with minimal disturbance of the flow field

被引:21
作者
Levitan, I
Helmke, BP
Davies, PF
机构
[1] Univ Penn, Inst Med & Engn, Vagelos Res Labs 1010, Philadelphia, PA 19104 USA
[2] Univ Penn, Dept Pathol & Lab Med, Philadelphia, PA 19104 USA
[3] Univ Penn, Dept Bioengn, Philadelphia, PA 19104 USA
关键词
hemodynamics; endothelial cells; electrophysiology; shear stress; biomechanics; biosensors; surface tension;
D O I
10.1114/1.1317529
中图分类号
R318 [生物医学工程];
学科分类号
0831 ;
摘要
An obstacle to real-time in vitro measurements of endothelial cell responses to hemodynamic forces is the inaccessibility of the cells to instruments of measurement and manipulation. We have designed a parallel plate laminar flow chamber that permits access to adherent cells during exposure to flow. The "minimally invasive flow device" (MIF device) has longitudinal slits (1 mm wide) cut in the top plate of the chamber to allow insertion of a recording, measurement, or stimulating instrument (e.g., micropipette) into the flow field. Surface tension forces at the slit openings are sufficient to counteract the hydrostatic pressure generated in the chamber and thus prevent overflow. The invasive probe is brought near to the cell surface, makes direct contact with the cell membrane, or enters the cell. The slits provide access to a large number (and choice) of cells. The MIF device can maintain physiological levels of shear stress (<1-15 dyn/cm(2)) without overflow in the absence and presence of fine instruments such as micropipettes used in electrophysiology, membrane aspiration, and microinjection. Microbead trajectory profiles demonstrated negligible deviations in laminar flow near the surface of target cells in the presence of microscale instruments. Patch-clamp electrophysiological recordings of flow-induced changes in membrane potential were demonstrated. The MIF device offers numerous possibilities to investigate real-time endothelial responses to well-defined flow conditions in vitro including electrophysiology, cell surface mechanical probing, local controlled chemical release, biosensing, microinjection, and amperometric techniques. (C) 2000 Biomedical Engineering Society. [S0090-6964(00)00210-1].
引用
收藏
页码:1184 / 1193
页数:10
相关论文
共 37 条
[11]   Molecular cloning and expression of a bovine endothelial inward rectifier potassium channel [J].
Forsyth, SE ;
Hoger, A ;
Hoger, JH .
FEBS LETTERS, 1997, 409 (02) :277-282
[12]   FLOW EFFECTS ON PROSTACYCLIN PRODUCTION BY CULTURED HUMAN-ENDOTHELIAL CELLS [J].
FRANGOS, JA ;
ESKIN, SG ;
MCINTIRE, LV ;
IVES, CL .
SCIENCE, 1985, 227 (4693) :1477-1479
[13]   INDUCTION OF HUMAN VASCULAR ENDOTHELIAL STRESS FIBERS BY FLUID SHEAR-STRESS [J].
FRANKE, RP ;
GRAFE, M ;
SCHNITTLER, H ;
SEIFFGE, D ;
MITTERMAYER, C ;
DRENCKHAHN, D .
NATURE, 1984, 307 (5952) :648-649
[14]  
FUNG YC, 1990, BIOMECHANICS MOTION, P418
[15]   FLOW-INDUCED CALCIUM TRANSIENTS IN SINGLE ENDOTHELIAL-CELLS - SPATIAL AND TEMPORAL ANALYSIS [J].
GEIGER, RV ;
BERK, BC ;
ALEXANDER, RW ;
NEREM, RM .
AMERICAN JOURNAL OF PHYSIOLOGY, 1992, 262 (06) :C1411-C1417
[16]   STRETCH-ACTIVATED SINGLE ION CHANNEL CURRENTS IN TISSUE-CULTURED EMBRYONIC CHICK SKELETAL-MUSCLE [J].
GUHARAY, F ;
SACHS, F .
JOURNAL OF PHYSIOLOGY-LONDON, 1984, 352 (JUL) :685-701
[17]   IMPROVED PATCH-CLAMP TECHNIQUES FOR HIGH-RESOLUTION CURRENT RECORDING FROM CELLS AND CELL-FREE MEMBRANE PATCHES [J].
HAMILL, OP ;
MARTY, A ;
NEHER, E ;
SAKMANN, B ;
SIGWORTH, FJ .
PFLUGERS ARCHIV-EUROPEAN JOURNAL OF PHYSIOLOGY, 1981, 391 (02) :85-100
[18]   Direct observations of the mechanical behaviors of the cytoskeleton in living fibroblasts [J].
Heidemann, SR ;
Kaech, S ;
Buxbaum, RE ;
Matus, A .
JOURNAL OF CELL BIOLOGY, 1999, 145 (01) :109-122
[19]   Rapid displacement of vimentin intermediate filaments in living endothelial cells exposed to flow [J].
Helmke, BP ;
Goldman, RD ;
Davies, PF .
CIRCULATION RESEARCH, 2000, 86 (07) :745-752
[20]   SHEAR ACTIVATED CHANNELS IN CELL-ATTACHED PATCHES OF CULTURED BOVINE AORTIC ENDOTHELIAL-CELLS [J].
JACOBS, ER ;
CHELIAKINE, C ;
GEBREMEDHIN, D ;
BIRKS, EK ;
DAVIES, PF ;
HARDER, DR .
PFLUGERS ARCHIV-EUROPEAN JOURNAL OF PHYSIOLOGY, 1995, 431 (01) :129-131