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Jul 19, 2018

Preparation of Post Staining Solutions

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Post staining of thin sections is used to increase contrast in TEM images. Typically this is accomplished in a two-step process using uranyl acetate, or a uranyl replacement, followed by a lead citrate solution.

 

View the Kirsch Note here

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  • Greetings I have been facing a problem with Technovit 9100. The polymerization never worked with me I have tried multiple times with the (stable and destabilized) basis solution in different conditions, however, I still end up with a liquid form which never hardens. Tissue type: Human acetabular bone with soft tissue attached. Total size: 13*10 mm.Human femoral head. Total size 5*5 mm. Protocol: 1) Dehydration on tissue rocker for (agitation): (50%, 70%, 80%, 95%, 95%) 1 hour each; (abs alcohol) overnight then (abs alcohol) for 1 hour. At room temperature 2) Intermedium on tissue rocker for (agitation): Xylin for (1 hour) then again Xylin for (1 hour). At room temperature 3) Pre-infiltration solution; Solution preparation: 100 ml of basis solution + ½ g Hardener 1. Solution used on tissue rocker for (agitation): 1 Hour At room temperature 4) Infiltration solution; Solution preparation: 100 ml of basis solution + 1.5g Hardener 1 + 10g of powder. Solution used for 24 Hour At 4C 5) Polymerization: Solution preparation: Stock A (100ml): 16g of powder + 80ml basis solution (mix until dissolve) + 0.8g Hardener 1 then top up the solution until 100 ml is reached. Stock B (100ml): 0.8 ml Hardener 2 + 0.4 ml Regulator + 100 ml basis solution Finally, the polymerization solution is used in a ratio of 9 parts from stock A + 1 part of stock B mixed immediately before use. We have tried a number of conditions to see if the polymerization will happen: Solution with the tissue at (RT, 4C and - 15C) for 24HSolution with the tissue at (RT, 4C and - 15C) for 72HSolution without the tissue at (RT, 4C and - 15C) for 24H However, all of that ended unsuccessfully. I would really appreciate any insight or help to this matter Sincerely Abdulaziz
  • When working with solutions for biological applications, distilled or deionized water is usually the solvent of choice. View the Kirsch Note here