Navigation Links
Optimizing electroporation parameters

Other important parameters for optimizing electroporation.
Influence of temperature

The temperature has a direct effect on the permeation voltage of the cell membrane (see field strength) as well as on the regeneration of the membrane following electroporation.

a) Influence of the temperature on the permeation voltage of the cell membrane:
Since the permeation voltage at 4C is twice that at room temperature (1), it is essential to take the temperature into account when determining the optimal field strength of the pulse. During electroporation at 4C, the necessary field strength of the pulse is nearly twice as high as those values for room temperature. However, mammalian cells are usually electroporated at room temperature.

b) Influence of the temperature on the regeneration of the cell membrane:
Incubating cells following electroporation at low temperatures (e.g. 4C) slows down the healing process of the cell membrane. In the case of eukaryotic cells, the resealing of the membrane pores can take half an hour or longer under these conditions. With certain cell types, this can lead to an increase in the amount of transfection material absorbed. However, some cells are extremely sensitive to low temperatures, particularly when permeated, and can suffer from irreversible damage after short incubation times in a cold environment. In those cases where electroporation at 4C leads to higher transfection rates, the cells' chances of survival can be boosted if they are resuspended in electroporation buffer at 37C or room temperature, cooled down to 4C and then transferred into precooled cuvettes. Following electroporation, the cells are cooled on ice for a maximum of two minutes and then heated to 37C in a water bath. Electroporation at higher temperatures (e.g. 25C) causes the permeated membrane areas to seal up more rapidly, which accelerates membrane regeneration and thus increases the cell survival rate. However, the transfection rate may be lower than that obtained when electroporation is carried out at low temperatures.

c) Influence of the temperature on the conductivity of the buffer:
The temperature has a profound effect on the conductivity of the electroporation buffer. Increasing the temperature causes the conductivity of a solution to increase as well, which may lead to lower transfection rates. For this reason, it is advisable not to work at temperatures in excess of 33C

(1) Zimmermann U. (1996) Effect of high intensity electric field pulses on eukaryotic cell membranes. In: Electromanipulation of Cells. U. Zi mmermann and G. Neill, editors. pp. 1. CRC Press, Boca Raton, 1996.



Page: All 1 2 3

Related biology technology :

1. Optimizing Transfection Conditions for Studying Signal Transduction Pathways
2. Optimizing pfuturbo DNA Polymerase Amplification Reactions with Perfect Match PCR Enhancer
3. Optimizing electroporation parameters
4. Optimizing Büchi® Rotary Evaporator Applications
5. Optimizing electroporation parameters
6. Optimizing Büchi® Rotary Evaporator Applications
7. Optimizing Bchi Rotary Evaporator Applications
8. Optimizing electroporation parameters
9. Optimizing electroporation parameters
10. Optimizing electroporation parameters
11. Optimizing electroporation parameters
Post Your Comments:

(Date:11/24/2015)... Tampa, Florida (PRWEB) , ... November 24, 2015 ... ... its biggest event of the year and one of the premier annual events ... USA, and ran from 8–11 November 2015, where ISPE hosted the largest number ...
(Date:11/24/2015)... , November 24, 2015 ... new market research report "Oligonucleotide Synthesis Market by Product ... (PCR, Gene Synthesis, Diagnostic, DNA, RNAi), End-User (Research, Pharmaceutical ... published by MarketsandMarkets, the market is expected to reach ... in 2015, at a CAGR of 10.1% during the ...
(Date:11/24/2015)... 2015 SHPG ) announced today that ... Jaffray 27 th Annual Healthcare Conference in New ... 8:30 a.m. EST (1:30 p.m. GMT). --> SHPG ) ... participate in the Piper Jaffray 27 th Annual Healthcare Conference ... December 1, 2015, at 8:30 a.m. EST (1:30 p.m. GMT). ...
(Date:11/24/2015)... 2015 /PRNewswire/ - Aeterna Zentaris Inc. (NASDAQ:  AEZS) ... remaining 11,000 post-share consolidation (or 1,100,000 pre-share consolidation) ... B Warrants") subject to the previously disclosed November ... 2015, which will result in the issuance of ... the issuance of such shares, there will be ...
Breaking Biology Technology:
(Date:11/10/2015)... , Nov. 10, 2015  In ... on the basis of product, type, application, ... included in this report are consumables, services, ... report are safety biomarkers, efficacy biomarkers, and ... this report are diagnostics development, drug discovery ...
(Date:11/9/2015)... DUBLIN , Nov. 09, 2015 /PRNewswire/ ... announced the addition of the "Global ... to their offering. --> ... "Global Law Enforcement Biometrics Market 2015-2019" ... Research and Markets ( ) ...
(Date:11/4/2015)... New York , November 4, 2015 ... to a new market report published by Transparency Market ... Share, Growth, Trends and Forecast 2015 - 2022", the global ... of US$ 30.3 bn by 2022. The market is ... the forecast period from 2015 to 2022. Rising security ...
Breaking Biology News(10 mins):