Whole mounts

WHOLE MOUNTS

1. WHOLE MOUNTS

2. Whole mount Objects which are thin, flat and small which do not require sectioning to reveal their structure are mounted entire is called Whole mount Example : filamentous algae , fungi fern prothallus,leaf epidermis,small flowers etc….. Whole mount preservation is adopted for a wide range of materials like whole plant ,stem ,leaves , flowers and fruits. This helps to retain the natural colour form and shape and also prevents decay

3. Importance of whole mount Whole mount preserve the specimens without disturbing their natural habit In the case of fern prothallus, reproductive bobies of algae & fungi etc ….their whole mount preparations give more lucid & clear structure than those of their sections

4. Methods of whole mounting 1 ) The Hygrobutol method used to prepare permanent whole mount of filamentous algae like ulothrix , spirogyra etc… steps :- a) Kill & fix filamentous algae in chrom-acetic acid fluid b) Wash the filament in running water . c) Stain using haematoxylin or carmine. d) Dehydrate using 15 , 30, 45, 70, 85% alcohol with long duration in last stage . e) Counter stain with erythrosine B / orange G / fast green. f) Wash & kept in 95% alcohol

5. Add gradual changes of hygrobutol till the mixture contains 90% hygrobutol & 10% alcohol Add 10 times diluted canada balsm with hygrobutol & allow balsm to evaporate till it becomes ready for mounting 2) The Dioxan method Very expensive method in which pure dioxan is used . only aqueous / very weak alcoholic stains are to be used .

6. Steps : - killing & fixing  staining  washing dehydration – using different changes of dioxan infiltrate with balsm diluted with dioxan  allow to evaporate  mount in balsm on a slide with cover slips . 3) The Creo sote method One superior method to Dioxan method. Give more transparency to thick materials like styles ,leaf epidermis etc….. Usually Beechwood Creosote is used in this method.

7. Steps :- Killing & fixing b) Staining c) Washing Pass the material through 50,70,80% ethyl alcohol for dehytration for 10 minutes in each stage e) Transfer material to a mixture of creosote & 85% alcohol. Pass through changes having gradual increase of creosote until it reaches pure creosote. Mount in balsm ( because the rate of evaporation for monuting consistency is very slow for creosote)

8. 4) The glycerin- Xylol method Killing Fixing Staining---using stains like haematoxylin Put material in 10% glycerine & allow this to evaporate for 4 days until glycerine becomes concentrated. e) Remove glycerine by changes of 95% alcohol.

9. Counter stain with acid stains, if reguired. Dehydrate completely using 100% alcohol. h) Pass through changes of alcohol, xylol mixture until reaches pure xylol.(each stage needs 5-10 minutes) Transfer to balsm diluted with xylol. Allow xylol to evaporate & mountng is done in balsm

10. 5 . The Vevetian turpentine method The material is stained & dehydrated by the glycerine evaporation method. Steps :- 1) remove the glycerine in the material by 95% alcohol.Give three changes in absolute alcohol. 2)Transfer the material to mixture of 10% venetian turpentine in absolute alcohol. 3) keep for 5 minutes.

11. 4) Then allow the alcohol to evaporate when the medium contains only turpentine . And mount the material in a drop of turpentine on a clean slide. 6) Put a cover slip over the material and keep flat to harden. 7) Seal the cover glass using a standard adhesive like DPX or canada balsam.

12. Permanent whole mount Methods : - Filamentous algae like Spirogyra, Oedogonium – First killed in any fixing solution. Then the excess killing solution is washed off with water . The material stained with a self- mordanting haematoxylin for about half to 1 hour

13. The stain is now washed off & the material transferred to a destaining solution of 0.1% hydrochloric acid in a cavity block. Stir well & then drain out . Then wash in tap water & examine with a microscope The washing is repeated until the nucleus & pyrenoids alone retain the colour

14. Temporary & Semi – permanent Whole mount slides A very simple method of preserving small filamentous algae. Placing it in a drop of 10% glycerine & covering with a cover slip -- purely Temporary method.. An improved preservative & mounting medium is the Lactophenol mountant. In filamentous algae Spirogyra ,ulothrix etc,….& fungi like Rhizopus, Aspergillus,phytophtora etc ….. May be mounted in this medium .

15. Aman’s Lactophenol phenol ---------20ml Lactic acid------20ml Glycerine--------40ml Water ----------20ml To mounting in this medium the material may be stained in cotton blue or aniline blue , The excess stain may be removed by giving a wash in the liquid it self .

16. Glycerine jelly components of glycerine jelly Gelatin - 5gm Water - 30ml Glycerine - 35ml Phenol dissolved in 10 drops of water --5 gm gelatin is dissolved in luke warm water. The other components are added to it & then filtered hot .it is kept in a closed bottle. Material like filamentous algae & fungi are stained in Haematoxylin & then dehydrated before mounting in the glycerine jelly .for dehydration.. the glycerine evaporation method is suggested.

17. A small quantity of glycerine jelly is placed on a slide & melted . Then the material to be mounted is removed from the glycerine & transferred into the warm jelly on the slide. A clean cover slip is now put over the jelly .& pressed gently to extrude the excess jelly. After cooling , the excess jelly around the cover slip is wiped off

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