[Pollinator] Recipe for Calberla's Fluid

[Peter Kevan]

Hi Peter! These recipes and discussion of methods are presented in the book edited by Dafni et al. Practical Pollination Biology (600 or so pages) which can be obtained in the USA through BioQuip and on Amazon. In EU it is available through Natural History Books (NHBS) and in Canada (and elsewhere) directly from the printers, Volumesdirect.com (or ?.ca). Cheers, Peter 

----- Original Message -----

From: "Peter Bernhardt" <bernhap2 at slu.edu> 
To: "Bee United" <beemonitoring at yahoogroups.com>, "Pollinator List-serv" <pollinator at lists.sonic.net> 
Sent: Monday, October 13, 2014 10:11:43 AM 
Subject: [Pollinator] Recipe for Calberla's Fluid 


We've received another request for a recipe for Calberla's fluid to stain pollen carried by bees and other pollinators. There is no reason to use a Safranin stain at al if you are using any form of light microscroscopy. Safranin sucks as it invariably gets into pollen cytoplasm obscuring exine apertures needed for identification. Restrict your staining to Calberla's fluid with a basic fuchsin base. 


Mother Bernhardt's Calberla's Solution 


5 ml glycerin 
10 ml of 95 or 100% ethanol 
15 ml distilled water 
1 - 3 drops of home made aqueous basic fuchsin based on a super saturated slurry* 


*The trick is to start by making a super-saturated solution of basic fuchsin and water (a slurry) and then filter the solution to remove the crystals. The highly concentrated solution of basic fuchsin can now be bottled and stored indefinitely (no refrigeration needed). 



Now, add the first three ingredients. When staining the solution of ethanol and water add the basic fuchsin solution drop-by-drop until it turns pink and you can still see your finger on the opposite side of the bottle. Think of the pink in a stained glass window. If the solution turns ruby, blood, or brick red you've over done it. 


I've also been asked about pollen atlases for the midwest. There are none. Dr. David Bogler started an online pollen atlas of the Midwest via the Missouri Botanical Garden. it was not finished due to lack of funding but he continues to work on it. Therefore, it's my advice you make a library collection of pollen slides of known species and photograph them as you go (at different angles and magnifications). 


Peter Bernhardt, Prof. of Biology 
Dept. of Biology 
Saint Louis University 
Saint Louis, MO 63103 
Tel. (office): 314-977-7152 


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