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Gabriella Turek
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Continued Operation sub pages fixes
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‎Beads.md

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This page is about fluorescent beads, used as fiduciary markers for
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multi-view registration in light sheet microscopy.
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This page is about fluorescent beads, used as fiduciary markers for multi-view registration in light sheet microscopy.
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## Estapor Fluorescent Microspheres
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- [website](http://www.estapor.com/estapor/en/fluorescent_microspheres/fluorescent_microspheres/19.html)
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- [website](https://www.estapor.com/estapor/en/fluorescent_microspheres/fluorescent_microspheres/19.html)
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- stock concentration for SPIM: 14 µl 0.5 µm beads in 14 ml medium
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(sample dependent, e.g. PBS/T for fruit fly, E3 for zebrafish)
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### tips & tricks
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### Tips & tricks
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- extended vortexing and/or treatment in ultrasonic bath is crucial
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for homogeneous bead distribution and successful bead detection
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- extended vortexing and/or treatment in ultrasonic bath is crucial for homogeneous bead distribution and successful bead detection
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- evaluate the bead concentration in agarose w/o specimen
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### F-XC050
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- "green" emission (see spectra)
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- EX 488 nm
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- appliations:
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- fiduciary marker in "green" detection channel for bright "green"
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fluorescence
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- fiduciary marker in "green" detection channel for bright "green" fluorescence
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- evaluation/adjustment of red-green overlay
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### F-Y050
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- "yellow" emission (see spectra)
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- EX 488/561 nm
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- applications:
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- fiduciary marker in "green" detection channel for weak "green"
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fluorescence
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- fiduciary marker in "red" detection channel for "green"
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fluorescence
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- fiduciary marker in "green" detection channel for weak "green" fluorescence
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- fiduciary marker in "red" detection channel for "green" fluorescence
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### spectral properties
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### Spectral properties
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[File:Estapor-beads.png|emission](File:Estapor-beads.png%7Cemission)
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spectra
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{% include image src="Estapor-beads.png" width="100%" caption="Emission spectra" %}

‎Fep_tube_cleaning.md

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This protocol is an important part of sample embedding with FEP tubes,
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e.g. [Zebrafish embryo sample
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preparation](Zebrafish_embryo_sample_preparation "wikilink").
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This protocol is an important part of sample embedding with FEP tubes, e.g. [Zebrafish embryo sample preparation](Zebrafish_embryo_sample_preparation).
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### material
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- FEP tubes (Bola S1815-04, inner diameter 0.8 mm, outer diameter 1.6
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mm)
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- FEP tubes (Bola S1815-04, inner diameter 0.8 mm, outer diameter 1.6 mm)
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- ultrasonicator
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- 1 M NaOH (Merck)
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- 0.5 M NaOH
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- double-distilled H2O
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- 70% ethanol
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- syringe filter (Millex-HV PVDF 0.45 µm)
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- B. Braun [Omnifix F Solo 1 ml
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Syringe](http://www.bbraun.com/cps/rde/xchg/bbraun-com/hs.xsl/products.html?prid=PRID00000578)
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- B. Braun [Omnifix F Solo 1 ml Syringe](https://www.bbraun.com/en/products/b/omnifix-f-solo.html)
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- B. Braun needle (100 Sterican, blunt)
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[File:Fep\_cable-drum.JPG|Bola](File:Fep_cable-drum.JPG%7CBola) S1815-04
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FEP tube [File:Syringe\_packed.JPG|B](File:Syringe_packed.JPG%7CB).
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Braun Omnifix F Solo 1 ml Syringe, B. Braun Sterican needle
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[File:Syringe\_unpacked.JPG|B](File:Syringe_unpacked.JPG%7CB). Braun
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Omnifix F Solo 1 ml Syringe w/ attached needle
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{% include image src="Fep_cable-drum.JPG" width="70%" caption="Bola S1815-04 FEP tube" %}
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{% include image src="Syringe_packed.JPG" width="70%" caption="Braun Omnifix F Solo 1 ml Syringe, B. Braun Sterican needle" %}
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### procedure
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{% include image src="Syringe_unpacked.JPG" width="70%" caption="Braun Omnifix F Solo 1 ml Syringe with attached needle" %}
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1. flush tubes with 1 M NaOH, use syringe with attached needle
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2. transfer flushed tubes to fresh Falcon with 0.5 M NaOH, use forceps
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3. put the Falcon in an ultrasonic bath for 10 min
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4. from now on, touch the tubes only with gloves and/or forceps\!
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5. transfer the tubes from the Falcon into small basin with ddH2O and
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flush them with ddH2O
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6. flush the tubes with 70% EtOH
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7. transfer the tubes to a fresh Falcon with 70% EtOH
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8. put the Falcon in an ultrasonic bath for 10 min
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9. transfer the tubes to a fresh Falcon with ddH2O for storage
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### Procedure
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## external links
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1. Flush tubes with 1 M NaOH, use syringe with attached needle
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2. Transfer flushed tubes to fresh Falcon with 0.5 M NaOH, use forceps
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3. Put the Falcon in an ultrasonic bath for 10 min
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4. From now on, touch the tubes only with gloves and/or forceps!
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5. Transfer the tubes from the Falcon into small basin with ddH2O and flush them with ddH2O
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6. Flush the tubes with 70% EtOH
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7. Transfer the tubes to a fresh Falcon with 70% EtOH
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8. Put the Falcon in an ultrasonic bath for 10 min
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9. Transfer the tubes to a fresh Falcon with ddH2O for storage
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- [Multilayer Mounting for Long-term Light Sheet Microscopy of
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Zebrafish](http://www.jove.com/video/51119/multilayer-mounting-for-long-term-light-sheet-microscopy-of-zebrafish)
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(external "JoVE" video article)
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## External links
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[Category:Operation](Category:Operation "wikilink")
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- [Multilayer Mounting for Long-term Light Sheet Microscopy of Zebrafish](https://www.jove.com/t/51119/multilayer-mounting-for-long-term-light-sheet-microscopy-of-zebrafish) (external "JoVE" video article)

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