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Eosin Staining Cells

Eosin Staining Cells - Histology Forum

Eosin Staining Cells - Histology Forum. Talk, discuss and post questions on general histology methods including tissue processing, embedding tissues, sectioning, special staining, enzyme staining and silver staining of samples.


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  #1  
Old 05-21-2008, 04:08 AM
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Default Eosin Staining Cells



Hi,
i cant find much information on Eosin.
Whats the difference between alcoholic Eosin and aqueous solutions?
Any advantages disadvantages?

thankx
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  #2  
Old 01-01-2015, 04:48 PM
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Default Re: Eosin Staining Cells

Viability of spermatozoa : fifteen minutes after sample collection, sperm viability is measured using the eosin-Y staining test that is obtained mixing 40 μl of an aliquot of spermatozoa with 20 μl of a eosin supravital stain (dissolve 0.5 - 67 g of eosin Y [C.I. 45380] in aqueous solution of 0.9% NaCl stored in a dark glass bottle at 4 C. Before use, the staining solution was brought to room temperature). The suspension was incubated for 30 sec at room temperature. Then, a 10 l droplet was transferred with the pipette to a microscope slide where it was smeared and allowed to dry in air. The slides were then stained in Harris' hematoxylin solution for 2 minutes and then washed for 30 sec with running cold tap water in order to remove the unbound nuclear stain. The next step involved 4-6 dips in acidic-ethanol solution (stock solution: 300 ml ethanol absolute + 100 ml dH20 + 2 ml HCl fuming 37%) to remove the non-specific bound dye (destaining) and then washed with running cold tap water for at least 2 minutes. The smears were air dried and examined directly. For each sample, 200 spermatozoa were assessed with microscope at a magnification of 1000x under oil immersion with a high-resolution 100x bright field objective and blue filter. Only sperm that were blue coloured were classified as alive and those that showed red or any reddish purple colouration were classified as dead. In alternative 1-2 min. after mixing 10 l spermatozoa suspension with 10 l of eosin supravital stain cover the sample with a coverslip and examined under a light microscope at 400-1000x magnification. Eosin dye penetrating head membrane of dead cells, which then have pink heads while the live cells appear unstained for membrane-intact (viable). Counting of living sperms (unstained) and dead ones (stained) on one hundred cells were randomly chosen in order to register the number of unstained mobile and immobile cells and stained spermatozoa (died cells). From this, the percentage of spermatozoa viability is estimated.
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Old 01-01-2015, 05:02 PM
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Default Re: Eosin Staining Cells

errata corrige two lines from the top : eosin supravital stain (dissolve 0.5 - 0.67g of eosin Y .......)
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Old 01-03-2015, 03:57 PM
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Default Re: Eosin Staining Cells

Harris' Hematoxylin Solution : dissolve in 1 liter dH2O, 22 g aluminum ammonium sulfate•12H2O (Fluka cod. 09693) and 7.5 g Hematoxylin. Then 0.75 g sodium iodate is added to oxidize the hematoxylin and the solution is mixed for 30 minutes and allowed to set overnight. All undissolved components are then filtered out and the solution is checked on a spectrophotometer at 560 nm, and additional dH2O is added until the absorbance reached 0.325, and then added 150 ml of trihydroxyhexane. Store reagent at room temperature (18-26 C) protected from light. Reagent is stable for 1 year that go discarded if staining time becomes excessive or if solution color changes from plum to blue or brown. Filter Harris Hematoxylin Solution before each use.
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