Posted on Leave a comment

Antibody Discoveries Customized for You

If you need immunohistochemistry for your project, we have years of experience working with both known and unknown antibody protocols.  We can get your project done fast with great results at a fraction of the price.  Call or email us today!

Picture of Mouse Stomach - Antibody HMGB1
Mouse Stomach – Antibody HMGB1
Posted on Leave a comment

Homemade Aqueous Mounting Media Works Great

The earlier 2 posts about the aqueous mounting media we made has a third update.  3 different labs to date has used our homemade aqueous mounting media with great success.  It has been used flawlessly on fluorescent antibody staining and special staining (oil red O) for histology techniques.

The current alternative to making it is buying it from 1 of many companies for an outrageous amount of money.  I wonder if one manufacturer makes large batches of it, then sells it to the different retailers?  What do you think?

The protocol needs a little tweaking as it’s very thick and I don’t think it needs to be.  There’s also the antifade agent (Either p-Phenylenediamine hydrochloride: 100 mg or n-propyl gallate: 500 mg) that is optional to add.  I did add it to the mix, using p-Phenylenediamine and it turned the liquid brown.  After doing this I and others had doubt about its effectiveness, but it worked and the brown color had no effect on the fluorescence or ORO.  I think next time I make it, I will use less antifade.

I remember making this stuff because it took so long, about 4 hours.  Of course, this was the first time and I was doing 10 other things besides this.

As promised, the recipe for this solution is copied here.  This is found on John A Kiernan’s website http://publish.uwo.ca/~jkiernan/aboutjk.htm, histology books and papers he has written or contributed to. Thank you John for giving us your insight into the world of histology, you have truly made difference to so many people!

Polyvinylpyrrolidone (PVP) medium

 

This is my favorite aqueous mounting medium. The composition can be varied according to each need (Kiernan, 1990). For immunofluorescence, make it up in a buffer and add an anti-fading agent.

 

Polyvinylpyrrolidone (M.W. 10,000): 25 g

Water (or a 0.1 M phosphate or

TRIS buffer, pH 7.4 or 9): 25 ml

Dissolve the PVP by leaving for several hours on a magnetic stirrer. Then add:

Glycerol: 1.0 ml

Thymol: One small crystal

Bottles of PVP medium usually keep for 2 to 3 years at room temperature. Keep it in a dark place if an anti-fading agent has been added. Discard if it looks infected or becomes too viscous.

Anti-fading agent:

Either p-Phenylenediamine hydrochloride: 100 mg

or n-propyl gallate: 500 mg

 

This mounting medium is more runny than glycerol jelly or Apathy’s. It is very easy to apply, and not prone to bubbles. The refractive index is 1.46 (Pearse, 1968), but increases as the water evaporates at the edges of the coverslip until unstained structures are barely visible. If you want a high degree of transparency, wait several days before sealing the edges of the coverslip.

 

 

  1. A. Kiernan

Department of Anatomy & Cell Biology

The University of Western Ontario

London, Canada N6A 5C1

 

Posted on Leave a comment

Aqueous Mounting Media, Update

Several labs have and continue to use the newly made aqueous mounting media.  So far it has been used for IF only but does do just as well as the commercially bought products. At this point the only difference that has been observed is the color of the liquid, it’s brown.  This does in no way cause any visual problems when viewing the slides under a fluorescent scope, causing no distortion or color skewing.

Amyloid fluorescent

I am hesitant to say this is good for all aqueous mounting types because it has not been used on special staining like oil red O.

I hope to try this on test slides within the next several weeks and will update again as this progresses.

Posted on Leave a comment

Small Business Histology

Come experience our extremely skilled and experienced technical team to show what your experiment results could look like.    Try us today.

background.jpg

 

 

 

 

 

 

An Expert Histology Service