How much powdered form of Wrights Stain and what liquid do I mix with it to make enough for 15 students?
Concentraton of the staining solution: 0.15% in methanol; shake well and leave to stand for 2 days or longer. Filter.
You need aprox. 500 µl/slide.
More information see in:J.A. Kiernan: Histological & Histochemical Methods: Theory & Practice
Hi, dear Heath,
it would be of help if you had included the task you are organizing (I guess for a certain course) for your students. If you are going to organize a seminar/lesson on "staining blood" [= smears, paraffin sections of e. g. bone marrow specimens, or what so ever] the I guess it might be less than you want your students to make stock or working solutions for further use in research or charity projects for the '3rd world'. There are a lot of "Wright's stains" (e.g. WRIGHT stain, WRIGHT &GIEMSA stain) out in the wild and therefore it is important to know which purpose the "stain powder" or "staining solution" finally should serve. Usually staining solutions of basic dyes like the cationic/basic NEUTRAL Wright's stain" (which classically is a mixture of an acid dye = eosin (red) and basic dyes = methylene blue dyes in methanol" are applied in a low concentration range, depending on the substrate to be stained (0.05-0.2% w/v, or 0.5-1%).
If you need to know about numbers/amount for 15 students then calculate - as a first attempt - 50 ml of solution / student x 15.... depending on which sample(s), which dispensing system [e.g. modern pipet for 100, 200, 500, 1000 µl - or just a simple single use/disposable 2.0 ml plastic pipet], how often and for which period of time they have to stain with such a solution.
Principally remember: Stock as well as working solutions - if they are made and stored appropriately - are durable (sometimes their staining properties increase with additional riping) and can be used up to 2 years and more.
Citation: "It is advisable, if possible, to keep a separate stock of Wright's stain for bone marrow staining which is kept at least 6 months before use.
Like a fine wine, the older Wright's stain gets, the better the quality and clarity of the final stain."
End of citation. source: from: https://www.medialabinc.net/spg448397/manual_staining_of_bone_marrow_preparations_wright.aspx.
If you need more and precise data/information on Wright's stains, either consult pertinent/relevant "(Hand)books" [usually in Histology or Histochemistry] like KIERNAN reading their recipes for Stainings/Stains OR alternatively, search GOOGLE for (e. g.)
|WRIGHT's stain basic dye | the first 20 out of nearly 5 mio results will help in finally calculating your demand on powder and/or liquid. Another possibility would be to search in the Q/A-archive of ResearchGate, just only searching for "Wright stain" (this producing an URL when browsing in the RG-platform: https://www.researchgate.net/search.Search.html?type=question&query=WRIGHT%27s++stain).
Characterisation of Wright's stain as only one example [to be found on: http://www.bu.edu/histology/m/append02.htm]: "
8. Wright's Stain
Wright's stain is a neutral stain produced by the interaction of an acidic and a basic dye, producing a large salt molecule with a colored dye in both its parts. The Romanovsky-type mixtures, (including Wright's and Giemsa stains) are the best known of these neutral stains and they are formed by the interaction of methylene blue and eosin.
With the Wright's stain, blood cells exhibit four major staining properties that allow the cell types to be distinguished. Basophilia (affinity for methylene blue), azurophilia (affinity for the oxidation products of methylene blue called azures, which are reddish purple), acidophilia (affinity for eosin), and neutrophilia (affinity for a complex of dyes in the mixture, which are pale lilac). In a stained blood smear, erythrocytes bind eosin and appear orange to pink, nuclei purplish blue, basophilic granules very dark bluish purple, eosinophilic granules red to red-orange, neutrophilic granules reddish-brown to lilac, platelets violet to purple, and lymphocyte cytoplasm stains pale blue." [http://www.bu.edu/histology/m/append02.htm]
Now I see that Ute Neubacher has helped you meanwhile anyway.... so ignore....
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