Chick and Quail Embryo (in ovo) – Neural Tube, Mesencephalon, Diencephalon (HH* Stage 10
For Chick and Quail Embryo (in ovo) – Neural Tube, Mesencephalon, Diencephalon (HH* Stage 10) applications we recommend the following electrodes:
Chick Limb EP:
- We usually recommend the following articles and electrodes used in those articles
- Congenic method in the chick limb buds by electroporation (Takayuki Suzuki* and Toshihiko Ogura, 2008)
- Electroporation into the Limb: Beyond Misexpression (Chapter 9: Takayuki Suzuki and Toshihiko Ogura)
- CUY612P5-5-3 (anode electrode) and CUY612P6-3 (cathode electrode) are used in the articles
- Technical drawing of both the CUY612P5-5-3 (anode electrode) and CUY612P6-3 (cathode electrode)
For your further information:
– Please note this recent NEPA21 paper (Nature) Specialized filopodia direct long-range transport of SHH during vertebrate tissue patterning.pdf
– It demonstrates In Ovo EP (forelimb)
– The publication is by one of our clients and he used our electrodes: the CUY611P7-2 (as the negative electrode) and the CUY611P3-1 (as the positive electrode)
– Technical drawings of both electrodes CUY611P7-2 and CUY611P3-1
Please also note:
– a recent presentation from Harvard Medical School
– The presentation records their success with our NEPA21 device (in comparison with the BTX device)
– It is Ex Ovo EP. They had previously used the BTX ECM830 before purchasing the NEPA21
Please note this comparison of the NEPA21 and BTX ECM830:
Measurement and Display of Delivered EP Pulse – Comparison of NEPA21 and BTX ECM830
It demonstrates the superiority of the NEPA21 device and the importance of controlling current rather than voltage.
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Please also note the following links and attached articles for further information on the NEPA21’s In Ovo capacity. (Please note where a reference is made in the resource material to the CUY21 systems (EDIT or SC), the NEPA21 replaces them and can replicate their exact same output pulse form – in addition to its own multi-pulse and polarity-exchange capability).
| CHICK EMBRYO: – Neural Tube – Limb Bud – Digestive Organ – Etc |
A useful place to start is this In Ovo EP Resource link on our website and the attached article titled: ‘Method of Electroporation for Early Chick Embryo’
Please note the following links to articles on our website and attached further resource material:
Cultured Embryos
– Publication and reference material:
- Manipulating gene expressions by electroporation in the developing brain of mammalian embryos
- NEPA21 and CUY21 Publication List and Electrode Recommedations for Rat Mouse Cultured Embryo
- NEPA21 and CUY21 Publication List for IN OVO EP – with Parameter Settings and Electrode Recommendations
- Electroporation-mediated gene transfer system applied to cultured CNS neurons
- Chapter 6: Method of Electroporation for the Early Chick Embryo (Jun Hatakeyama and Kenji Shimamura)
In Ovo – Neural Tube and Brain Regions
– Publication and reference material:
1. Gain- and loss-of-function in chick embryos by electroporation
2. Misexpression of genes in brain vesicles by in ovo electroporation
3. Applications for Chick Embryo
In Ovo and Ex Vivo and Ex Ovo Chick Retina EP
– Publication and reference material:
- Electroporation for early chick embryos using News culture (gastrula) – Application note 1.pdf
- Electroporation-mediated gene transfer system applied to cultured CNS neurons – Application Note 2
- Bimodal functions of Notch-mediated signaling are involved in neural crest formation during avian ectoderm development
- Irx4-mediated regulation of Slit1 expression contributes to the definition of early axonal paths inside the retina
- Ex ovo electroporation for gene transfer into older chicken embryos
- Electroporation-mediated gene transfer system applied to cultured CNS neurons
- Chick Embryo Retina EP.pptx – powerpoint (Attached)
Focal Electroporation on Slice Cultures and In Utero EP
– Publication and reference material:
1. For Slice Culture
Negative electrode: CUY700 series or CUY701 series
Positive electrode: CUY614 & 615, CUY611P3-1 or CUY195P0.3
Protocols:
2. For In Utero
Electrodes: CUY650P3 or CUY650P5
Protocols:
NOTE:
– The appropriate electrode size option is dependent on the size of your target.
– Kindly note that we are also able to custom-manufacture electrodes to your own specifications.
New Cultures
– Publication and reference material:
– Electroporation for early chick embryos using News culture (gastrula) – Application note 1.pdf
– Collinear activation of Hoxb genes during gastrulation is linked to mesoderm cell ingression
– Dual mode of paraxial mesoderm formation during chick gastrulation
Limb embryonic ectoderm
– Publication and reference material:
Please note the following articles, especially the first named article
- Efficient Ectopic Gene Expression Targeting Chick Mesoderm
- FGF8 initiates inner ear induction in chick and mouse
NOTE:
In the attachment Limb bud 2003.pdf, the article titled “Tbx5 and Tbx4 trigger limb initiation through activation of the Wnt/Fgfsignaling cascade”, Takeuchi et al, stage 13 is specifically mentioned.
Chick Limb EP:
For your further information: – Please note this recent NEPA21 paper (Nature) Specialized filopodia direct long-range transport of SHH during vertebrate tissue patterning – It demonstrates In Ovo EP (forelimb) – The publication is by one of our clients and he used our electrodes: the CUY611P7-2 (as the negative electrode) and the CUY611P3-1 (as the positive electrode) – Technical drawings of both electrodes CUY611P7-2 and CUY611P3-1
Please also note: – a recent presentation from Harvard Medical School – The presentation records their success with our NEPA21 device (in comparison with the BTX device) – It is Ex Ovo EP. They had previously used the BTX ECM830 before purchasing the NEPA21
Please note this comparison of the NEPA21 and BTX ECM830: Measurement and Display of Delivered EP Pulse – Comparison of NEPA21 and BTX ECM830 It demonstrates the superiority of the NEPA21 device and the importance of controlling current rather than voltage.
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Regarding the Parameter List The parameter list is similar in format to the following table:
There are 6 suggested condition-settings on the parameter list. It recommended that you try all 6 condition-settings first time out. Based on the results feedback you provide me after your experiments I will then suggest further optimised setting-conditions. Please note that we can repeat this process as often as you wish while you have the device. This is a process we have found very efficient in assisting other clients optimise their particular applications of interest.
To ensure early productivity in this endeavour it is vitally important (for me) that you record the data feedback I need in order to make appropriate optimisation suggestions. On the same line of each condition-setting, there is a section for you to record the measurements (Red) and result (Green) information. The ‘Measurement’ data is displayed on the face of the NEPA21 device after the delivery of each condition. The ‘Result’ data (Viability % and Transfection Efficiency % ) are estimate calculations I ask you to make based on your observation. It is very helpful if you can also send photos of these results. In addition and if merited, a short narrative observations on the results can prove useful. In short, the better the quality of information we have the easier it is for us to make appropriate optimisation suggestions.
For your information, please note these links to further information: – Down Load NEPA21 Presentation – Down Load Comparison of NEPA21 with Nucleofector Neon
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Something else for you to consider:
Follow this link to our webstie for In Ovo Limb Bud Sonoporation