2014/05/28 格蘭氏染色----染色試劑配置 (修編中)

Crystal Violet Stain  (Hucker modification)
 
       Solution A：Dissolve 2.0 gm of crystal violet (85% dye content ) in 20 ml of 95% ethyl alcohol
     
       Solution B：Dissolve 0.8 gm ammonium oxalate in 80.0 ml distilled water.

Mix solution A and B.

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Gram's Iodine ( Lugol's )

Dissolve 2.0 gm of potassium iodide in 300 ml of distilled water and then add 1.0 gm iodine crystal.


Iodine, 5% Aqueous Solution ( Ex. 30 )

Dissolve 2.0 gm of potassium iodide in 300 ml of distilled water and then add 2.0 gm iodine crystal.

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Malachite Green Solution ( spore stain )

Dissolve 5.0 gm of malachite green oxalate in 300 ml  distilled water.

((老師筆記：2.0g + 100 ml water →→無法全溶

→→  改用1g +100 ml  

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Safranin (for gram staining)

Safranin O (2.5% sol'n in 95% ethyl alcohol )..............10.0 ml
Distilled water ..........................................................100.0 ml

((摘自：Microbiological Applications p.329,330,331))


品牌： WAKO, Fisher  (查售價)


操作心得：

Safranin 2.5% 溶於 95% 酒精中，
預計配置1000 mL ( 所需 Safrnin O 為 100mL ) 總量為 1100mL
               
按化學重量百分比計算  X/X+100=25.6(5)
但以生物學角度操作，取 25g Safrnin O 溶於100mL水
(推導原理：只需染色，不至於到需要精準操作，目的：染色!!)

Iodine  顆粒難溶，碘為結晶粒。