Navigation Links
Optimizing electroporation parameters

Other important parameters for optimizing electroporation.
Field Strength The field strength (V/cm) of the electrical pulse used is an essential factor in determining the survival rate, as well as the transfection rate, of the cells used.

If the field strength of the pulse exceeds a specific threshhold (= critical external field strength), reversible permeation occurs in the cell membrane. This so-called permeation voltage is heavily dependent on the temperature at which electroporation takes place. The diagrams in Fig. 1 show the minimum values of the pulse voltage which have to be set in relation to the cell diameter and the temperature at which the electroporation is performed. The diameter of the cell is determined after the cells have been incubated in electroporation buffer for 1015 minutes (see adjustment of electroporation buffer). In addition, the gap width of the cuvettes must be taken into account when the minimum pulse voltage is determined. If the gap width is doubled, the pulse voltage must also be doubled in order to obtain the same field strength. A general rule when determining the ideal field strength is that small cells require a higher field strength in order to achieve membrane permeation. The pulse voltages in Fig.1 and the corresponding Table 1 are the minimum values at which the membrane can be permeated.

However, depending on the cell type used, optimal transfection efficiency is often only achieved at significantly higher voltages. To determine the optimal pulse voltage, it is advisable to carry out a series of experiments in which the minimum value, twice the value and then three times the value shown in Table 1 are used for suspension cells, and up to five times the value for adherent cells.

Cells which do not assume a rounded form in the electroporation buffer (see Fig. 2) often require even higher pulse voltages before optimal transfection can occur.

Please note that increasing the pulse voltage can increase the transfection rate but, at the same time, can also increase the cell mortality rate.

Cell diameter, in m Voltage 2 mm room temp. Voltage 4 mm room temp. Voltage 2 mm 4C Voltage 4 mm 4C
5 530 1100 1100
10 270 540 540 1100
15 180 360 360 710
20 130 260 260 530
25 110 220 220 430
30 90 180 180 360
35 80 160 160 310
40 70 140 140 270
45 60 120 120 240
50 50 100 100 210
60 40 80 80 160
80 30 60 60 120
Table 1. Minimum pulse voltages at which the cell membranes may be permeated, in relation to the diameter of the cell after a 1015 min incubation in the electroporation buffer, the electroporation temperature and the gap width of the cuvette. Note: Depending on the cell line, the optimal pulse voltage for the electroporation experiment may be between two and five times higher. Figure 1. Minimum pulse voltage at which the cell membrane is permeated. The minimum pulse voltage is dependent on the cell diameter following incubation in electroporation buffer, as well as on the temperature, and on the gap width of the cuvettes. The values shown can be used to determine the ideal pulse voltage to be set on the device.

Figure 2. Cells swell up under hypoosmolar conditions. In this state, electroporation can be performed under milder pulse conditions.

Example:

Cell type: Suspension cells

Cell diameter in electroporation medium: ~ 20 m

Gap width of cuvette: 2 mm

Temperature in the cuvette: Room temperature

Minimum pulse voltage according to diagrams (Fig. 1): ~ 130 V

Series of experiments for optimizing pulse voltage: 130 V / 260 V / 390 V



'"/>

Source:


Page: All 1 2 3 4 5 6

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:
*Name:
*Comment:
*Email:
TAG: Optimizing electroporation parameters

(Date:12/22/2014)... Dec. 22, 2014  ( www.competitivehealth.com ) — Competitive ... medical bill review and advocacy service, has signed an ... WellCard Savings discount health services marketplace. 63% ... more than they expected to pay. As part of ... costs, WellCard Savings is pleased to offer medical bill ...
(Date:12/22/2014)... 22, 2014 Fertility Associates of Memphis ... Bank USA , a network of more than 20 ... partnership, patients have fast access to a national pool ... IVF . Fertility Associates of Memphis is the only ... treatment option. , Dr. William Kutteh , director ...
(Date:12/19/2014)... BioPlus Specialty Pharmacy (BioPlus), one ... of Nick Maroulis, Pharm.D. to the newly created position ... In this position, Dr. Maroulis will continue to manage ... of our multi-site pharmacies as the company expands. Dr. ... that time he has served in many different capacities ...
(Date:12/19/2014)... Dec. 19, 2014 Bina Technologies, Inc. (Bina, ... that they have been acquired by Roche (SIX: RO, ... privately held company that provides a big data platform ... (NGS) data for the academic and translational research markets. ... and will continue to focus on development of their ...
Breaking Biology Technology:WellCard Savings Announces CoPatient Medical Bill Review Service for Members 2Fertility Associates of Memphis Announces Partnership with Donor Egg Bank USA 2Fertility Associates of Memphis Announces Partnership with Donor Egg Bank USA 3BioPlus Specialty Pharmacy Promotes Dr. Nick Maroulis to Vice President of Specialty Pharmacy Services 2Bina Technologies is acquired by Roche. 2Bina Technologies is acquired by Roche. 3
... Aligns Operations for Enhanced Performance, DECATUR, ... (NYSE: ADM ) chairman, chief executive ... organizational restructure of the,Company, designed to enhance ... responsibility for long-term success. The reorganization creates ...
... oximetry technology to significantly reduce, false alarms without missing ... ... Aug. 22 Masimo (Nasdaq:,MASI), the inventor of Pulse CO-Oximetry ... partnership with Apria,Healthcare to provide pulse oximetry products and accessories ...
... symbol: SBS, - Canadian Biotechnology Company confirms commercial ... AI Milano variant and clarifies status of native ... - SemBioSys Genetics Inc.,(TSX:SBS), a biotechnology company developing ... diseases, today updated a release,issued on July 12, ...
Cached Biology Technology:ADM Reorganizes Operations 2ADM Reorganizes Operations 3ADM Reorganizes Operations 4Apria Healthcare Chooses Masimo SET Pulse Oximetry for Use in the Company's Home Health Care Business 2Apria Healthcare Chooses Masimo SET Pulse Oximetry for Use in the Company's Home Health Care Business 3SemBioSys updates Apo AI development program 2SemBioSys updates Apo AI development program 3SemBioSys updates Apo AI development program 4
(Date:12/11/2014)... 2014  That blood pressure plays a role in human ... the medical term for high blood pressure – was first ... inflatable cuff that,s used in measuring blood pressure was invented ... new about hypertension, its triggers and its effects. In fact, ... and the best ways to treat it. For ...
(Date:12/10/2014)... 9, 2014  Valencell, a leader in performance biometric ... staggering demand from its licensees for highly accurate, clinically ... not solely coming from fitness and health sectors, but ... "A wearable is only as useful as the biometric ... ultimate driver in long-term mass consumer adoption of wearable ...
(Date:12/10/2014)... Research and Markets ( http://www.researchandmarkets.com/research/xclcvw/biometrics_market ... "Biometrics Market in Japan 2014-2018" report to ... The integration of biometrics and ... and upgradation of the driver,s license is one ... Besides the aforementioned projects, biometrics is being used ...
Breaking Biology News(10 mins):Research points to need for new approaches to treatment of high blood pressure 2Research points to need for new approaches to treatment of high blood pressure 3Research points to need for new approaches to treatment of high blood pressure 4Wearable Technology Products Demand Highly Accurate Biometric Technology 2Biometrics Market in Japan 2014-2018: Key Vendors are DDS, Fujitsu, Hitachi and NEC 2
... sole proprietors of their nests, some tropical species time ... a 150-year-old evolutionary mystery, says Queens University Biology professor ... confirms one of Charles Darwins more controversial theories ... many experts that different species can arise, unhindered, ...
... The Shifting Baselines Toolset is presented to the public and ... , When: Public symposium 1pm to 5:15pm, Wednesday 14th November ... city campus. , The meeting will be launched with a ... for interviews on the 14th and on other days by ...
... 13, 2007) A major study that will provide a ... be conducted by the La Jolla Institute for Allergy ... The five-year study, funded by the National Institute ... National Institutes of Health, could lead to revolutionary new ...
Cached Biology News:'Time-sharing' tropical birds key to evolutionary mystery 2How much fish have we got? 2How much fish have we got? 3NIH selects LIAI for major study on allergy molecular causes and possible treatments 2NIH selects LIAI for major study on allergy molecular causes and possible treatments 3NIH selects LIAI for major study on allergy molecular causes and possible treatments 4
Human ROR1 Affinity Purified Polyclonal Ab...
Ultra-sensitive, non-radioactive measurement of COX-I and COX-II activity...
Phospho-p70 S6 Kinase (T229) Affinity Purified PAb...
Recombinant Ovine (Sheep) Growth Hormone, Ultra Pure...
Biology Products: