Wednesday 17 February 2021

Triple Stain Wacker 3A

The Wacker 3A staining method (developed by Robin Wacker) is one of the methods which can be used to distinguish cell differences in plants. The images of the cross section of a Rubus idaeus (raspberry) sprout are showing a nice example:


The method is based on the following solutions: 
  • Astra blue (2% in distilled water)
  • Acriflavin (1% in distilled water) 
  • Acridine red (1% in 50% ethanol).  
*All solutions slightly acidified with 2ml acetic acid per 100ml solution.


For a less laborious and faster colouring, a colour mixture can be made from this, for simultaneous colouring according to Rolf-Dieter Müller. The composition of the W3Asim mixture is: 
  • Astra blue, Acriflavin and Acridine red in a ratio of 4: 1: 1. 
With its rich turquoise colour tone, the colour of not lignified tissues is very close to the colour effect of the original Wacker staining method. For details, consult the literature below:

  • Wacker, R. (July 2006). "Eine neue und einfache Methode zur polychromatischen An färbung von Paraffinschnitten pflanzlicher Gewebe für Durchlicht- und Fluoreszenzmikroskopie". Mikrokosmos, (4), 210- 212. [Online]

  • Müller, R.D., Weiß, J. (2011). "Wacker für alle - neue Simultanfärbungen auf Basis der W3A Färbung von Robin Wacker". [Online]

Rubus idaeus or raspberry is very suitable for growing in slightly moist places due to the construction of the leaves. These are felt-like on the bottom side, where the stomata are located and are not moistened by water. This allows the evaporation of water to continue regularly, as long as the state of the atmosphere allows this.

Raspberries develop special horizontal roots under the ground, on which buds develop at the upper side. The branches emerging from these buds form independent plants as soon as the common root dies. They are naturally arranged in rows and sometimes quite far away from the mother plant.

The raspberry is found in forests and rocky places in Central and Northern Europe. It is also widely cultivated.

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