Sonidel Limited

NEPA21: In Vivo Mouse EP – Cerebellum – Pups between 3-8 days old

In Vivo Neonatal Brain (Cortex / Hypothalmus) EP
–        You may use either the CUY650P7 or CUY650P10 electrode.
–        You will also need the C118 cable. (C118 Technical Drawing and Photo of connection)
–        The method is explained in the following article:

Microtubule-based nuclear movement occurs independently of centrosome positioning in migrating neurons

–        Photo Illustration to accompany the article above.
–        This method may be executed with or without the use of gauze (our preferred method is without gauze).
EP Condition sheets for both options are available to clients who have purchased the NEPA21 system.

While the ‘Ueshima et al’ method in the above ‘Microtubule-based’ article demonstrates P8, we believe the method will work just as well for P10-14.

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NEPA21: In Vivo Neonatal Mouse Brain EP – subventricular zone (SVZ) – Postnatal day 2 pups, with CUY650P10

Please note the advantages of knowing electric resistance and current as important EP Factors.

 

Sample A	Setting: 100V	==>	Impedance: 200Ω	==>	Output Ampere: 0.5A	The Results will
Sample B	Setting: 100V	==>	Impedance: 500Ω	==>	Output Ampere: 0.2A	not be the same

 Please note the following CUY21 publications:egory

	Title	Electrode	Voltage	P on	P off	No of Pulses
Rat and Mouse (Neonatal) Brain	Renaud et al. Plexin-A2 and its ligand, Sema6A, control nucleus-centrosome coupling in migrating granule cells Nature Neuroscience, Volume 11, Number 4, Pages 440-449, April 2008	CUY650P10	75V	​50ms	450ms	5
	Boutin et al. Efficient In Vivo Electroporation of the Postnatal Rodent Forebrain PLoS ONE, Volume 3, Issue 4, e1883, 2 April 2008	CUY650P10	100V	50ms	950ms	5
	Umeshima et al. Microtubule-based nuclear movement occurs independently of centrosome positioning in migrating neurons PNAS, Volume 104, Number 41, Pages 16182-16187, October 9, 2007	CUY650P7 or CUY650P10 Syringe needle	70V	50ms	150ms	6

 

For a single output-pulse setting (CUY21 mode), we recommend that you next test 100V, 90V, 80V and 70V (5 pulses, pulse duration 50ms and pulse interval 950ms).

EP Condition Sheets for the NEPA21 4-Step multi-pulse protocol are available to NEPA21 clients

 

Moreover, if the NEPA21 is compared with other competing devices for in-vitro cell transfection, we again observe significantly better results with the NEPA21 system.

Please note the following link:

Neurospheres – Neural Stem Cells Derived from Mouse SVZ, 24 hrs, 48 hrs and 72 Hrs after EP

Other Relevant Links:
–  Postnatal Cerebellum EP with a 3 part electrode (CUY699P7x6) or equivalent electrode combination
–  Ex vivo and In Vivo (Retina)
Postnatal (P0) EP – Mouse Retina Ex Vivo EP
We recommend the CUY650P7 electrode.
Please note the following;
For Methodology, please note the publication: Efficient In Vivo Electroporation of the Postnatal Rodent
–  Mouse Embryo – Retina/Eye