eprintid: 8976
rev_number: 5
userid: 6
dir: disk0/00/00/89/76
datestamp: 2011-11-10 19:58:10
lastmod: 2016-11-15 13:48:33
status_changed: 2011-11-10 19:58:10
type: thesis_degree
metadata_visibility: show
contact_email: bal46@pitt.edu
item_issues_count: 0
eprint_status: archive
creators_name: Lambie, Bradley Alan
creators_email: bal46@pitt.edu
creators_id: BAL46
title: Making Single-cell Electroporation with Microelectrodes Predictable and Reproducible.
ispublished: unpub
divisions: sch_as_chemistry
full_text_status: public
keywords: carbon fiber etching; current interruption; electroporation; microelectrodes; single-cell electroporation
abstract: Electroporation is the creation of transient pores in a membrane by the application of an external electric field. When using microelectrodes, which can be used to electroporate single-cells, for applying an electric field to the cell, there is a question of how much voltage to apply. Unlike in bulk electroporation where hundreds of volts may be applied between electrodes, a rather small voltage is applied to a microelectrode in single-cell electroporation. In the single-cell experiment with microelectrodes, a substantial fraction of the voltage does not exist in solution because it is lost at the microelectrode/solution interface. This problem is the same as the classical electrochemist's problem of knowing the 'iR' drop in solution and correcting for it to obtain true interfacial potential differences. Therefore, we have used current interruption experiments to determine the iR drop in solution near microelectrodes. Because the electric field produced by microelectrodes is inhomogeneous, computer simulations were preformed to understand the electric field distribution. Results of the current interruption are validated by comparing two independent measurements of the resistance in solution: one value results from the measured iR drop in conjunction with the known applied current. The other value results from a measured solution conductivity and a computer simulated cell constant. This paper shows how to calculate the approximate current required to electroporate a cell with a microelectrode of a particular size, shape and distance from the cell. Carbon fiber microelectrodes were used to electroporate single A549 cells using the current calculated.
date: 2010-09-30
date_type: completed
institution: University of Pittsburgh
refereed: TRUE
etdcommittee_type: committee_chair
etdcommittee_type: committee_member
etdcommittee_type: committee_member
etdcommittee_type: committee_member
etdcommittee_name: Weber, Steve
etdcommittee_name: Michael, Adrian
etdcommittee_name: Orwar, Owe
etdcommittee_name: Amemiya, Shigeru
etdcommittee_email: sweber@pitt.edu
etdcommittee_email: amichael@pitt.edu
etdcommittee_email: amemiya@pitt.edu
etdcommittee_id: SWEBER
etdcommittee_id: AMICHAEL
etdcommittee_id: AMEMIYA
etd_defense_date: 2010-03-26
etd_approval_date: 2010-09-30
etd_submission_date: 2010-08-07
etd_access_restriction: 5_year
etd_patent_pending: FALSE
assigned_doi: doi:10.5195/pitt.etd.2011.8976
thesis_type: dissertation
degree: PhD
committee: Steve Weber (sweber@pitt.edu) - Committee Chair
committee: Adrian Michael (amichael@pitt.edu) - Committee Member
committee: Owe Orwar () - Committee Member
committee: Shigeru Amemiya (amemiya@pitt.edu) - Committee Member
etdurn: etd-08072010-140815
other_id: http://etd.library.pitt.edu/ETD/available/etd-08072010-140815/
other_id: etd-08072010-140815
citation:   Lambie, Bradley Alan  (2010) Making Single-cell Electroporation with Microelectrodes Predictable and Reproducible.  Doctoral Dissertation, University of Pittsburgh.    (Unpublished)  
document_url: http://d-scholarship-dev.library.pitt.edu/8976/1/BLambie2010.pdf