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FEP Tube Cleaning |
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FEP Tube Cleaning |
This protocol is an important part of sample embedding with FEP tubes, e.g. Zebrafish embryo sample preparation.
- FEP tubes (Bola S1815-04, inner diameter 0.8 mm, outer diameter 1.6 mm)
- ultrasonicator
- 1 M NaOH (Merck)
- 0.5 M NaOH
- double-distilled H2O
- 70% ethanol
- syringe filter (Millex-HV PVDF 0.45 µm)
- B. Braun Omnifix F Solo 1 ml Syringe
- B. Braun needle (100 Sterican, blunt)
{% include image src="Fep_cable-drum.JPG" width="70%" caption="Bola S1815-04 FEP tube" %}
{% include image src="Syringe_packed.JPG" width="70%" caption="Braun Omnifix F Solo 1 ml Syringe, B. Braun Sterican needle" %}
{% include image src="Syringe_unpacked.JPG" width="70%" caption="Braun Omnifix F Solo 1 ml Syringe with attached needle" %}
- Flush tubes with 1 M NaOH, use syringe with attached needle
- Transfer flushed tubes to fresh Falcon with 0.5 M NaOH, use forceps
- Put the Falcon in an ultrasonic bath for 10 min
- From now on, touch the tubes only with gloves and/or forceps!
- Transfer the tubes from the Falcon into small basin with ddH2O and flush them with ddH2O
- Flush the tubes with 70% EtOH
- Transfer the tubes to a fresh Falcon with 70% EtOH
- Put the Falcon in an ultrasonic bath for 10 min
- Transfer the tubes to a fresh Falcon with ddH2O for storage
- Multilayer Mounting for Long-term Light Sheet Microscopy of Zebrafish (external "JoVE" video article)