By Tiwari, G. S. Hoondal
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Additional resources for Laboratory Techniques In Microbiology & Biotechnology
2. Heat fix the smear and treat the smear for 10 min. with methylene blue stain. 3. Wash the slide under running tap water and stain the smear with carbol fuchsin for a shol1 while (5 sec) and wash immediately with water. 4. Blot dry and examine under oil immersion objective. Live vegetative cells appear purple and dead are stained red or pink. Live spores take up faint pink color and dead take up hilic color. Questions I. What is the concentration of methylene 'blue or carbol fuchsin? 2. What is intravital stain?
Iii)Neutral red solution: for modified Neisser method - Mix 1 g neutral red and 2 ml 1% glacial acetic acid in 1000 ml distilled water. Loeffler method Loeffler's methylene blue stain Procedure Albert method I. Prepare bacterial smear on clean grease free slide, air dry and heat fix it. 2. Stain it with Albert stain for 5 min. 3. Wash the stain with running tap water. Add Albert's iodine for Imin. 4. Wash with water, blot dry, and examine under oil immersion objective. Record the observations.
Now add more Ag NO, solution drop wise until precipitates returns and does not dissolve) Mordant (phenol I g and tannic acid 5g dissolved in 100 ml distilled water) Becker's method a. b. Fixative and mordant same as above. Stain: Basic fuchsin saturated alcoholic solution 45 ml mixed with 18 ml Shunk's mordant (spirit 18 ml and aniline oil 4 ml) and volume made to 100 ml with distilled water. Procedure Fontana's method I. 2. 3. 4. 5. 6. 7. Scrap material from the teeth using sterile needle. Transfer the material to slide and spread this on to the slide to prepare a thin smear.
Laboratory Techniques In Microbiology & Biotechnology by Tiwari, G. S. Hoondal