Books QQ BB) co Asttomunry ~ —vtereview Tablet Some common Seater BH in res ot Moteadar Pando bas Concerton Chemie Se” nd Lanta eR Sonn ed ‘Bae aan esi wit os 28% aa) Boh anda, Wen ect ait oes 238%00) Both Acctoce 370th) Mice Gtioreocm iss 734) Mice eomie toot 13 thmaqy Mice Dionne ‘1 Mire Etnasol Be saute) Bate OTRO cerncurnes starrn tae Books @)Q BPE) co) atten = | vitereview « ™ Ses @ Micure 2 Mice m aor Mare 71262 tube) More a Mt o 67 ate) Mico 25 Stdragy Both 2 13 tat) Mixture Sites 28aq) Mature Poussim deromae 238 S80) Mire = ort Mur Propione Todor acd = 1340 1.241%) Both a62.6 Bap /tetangeci Google —erromosomes sharma sharma Ee « Books QQ BB) co Asttomunry ~ —vtereview Alcoholic Carnoy’s fuid (1886) Gioia wei acd pat Sccetaacl 3 ans Is effective forall plant, animal and human materials both for squash and block preparations, the period of fixation varying from 15 min to 24 h in cold or at room temperature. The fluid should be washed out with 70 or 90 per ent ethanol, Difficult materials can be mordanted in ethanol: 1-3 percent ferric ammonium sulphate (7:25) mixture for 3-12 h after fixation (Lesins, Google —_evomosomes sharma shama Ew = Books QQ B BK co Aadtomytorary * ——_Wte review cenr 1954). Certain Rhodophyceze are mordanted in aqueous 0.5-S per cent ferric ammonium sulphate solution after fixation (Austin, 1959). Modifications include mixtures in the proportions 1:1 (Von Beneden and [Neyt, 1887) and 1:2. Zacharias (1888) added a few drops of osmic acid solution to acetic aeid-ethanol (1:4) mixture, Burns and Yang (1961) fixed ‘Nicotiana microspores in acetic acid-95 per cent ethanol mixture (3:2) for 15 min after dissolving the pollen grain walls. For electron microscopic studies, anthers can be fixed in a mixture of eadmium chloride and absolute ethanol for 30 min or in only 0.5 m CdCl, for 10 min followed by refixation in acetic-ethanol (1:3). A modiication containing acetic acid, 96 per cent ethanol, concentrated HCl and distilled water (:3:2:2) has been used for mosquito chromosomes after 30s in Carnoy’s fixative (Amirkhanian, 1968). Carnoy’s fluid II (1886)LL 1€ > CB ie lokagoge canickd-cighagenisny-PMrelpg-DMPERdy-cromorona: shar shrmatou-Bci- TODA 5S Google eromosomes sharma sama Books QQ BB) co Asttomunry ~ —vtereview Carnoy's fluid IL (1886) Ghilnaieadd pat 1 Sou Hie See Fae {tis widely used for animal and human tissues and for flower buds. The period of treatment is from 15 min to 24h in cold oF in room temperature Metzger and Leng (1955) modified it by saturating it with HgCl, for certain Teguminous plants. Another modification, with ingredients in the proport fof 1:3:4 and 1:1:3 (Semmen’s fluid), has been used in Compositae and ‘other families (Turner, 1956) The fluids, mixed in equal proportions (1:1:1) Ihave been used on certain Heteroptera Carnoy and Lebrun’s Fluid (1887) luc aceic acid pact Google _ctromosomes sharma sharma isa Books ae 22 Aadtomyttray = Whterevew poss <> Ry ‘A modification by G. S. Sansom contains Gla seic acd pat 1 Ehlers Shim Stoic stapel 13 art CComoate nana to wtraon ‘The mixture is prepared just before use and is successful for insect ovum with shel and for certain vertebrate materials. It penetrates very rapidly, the jon being fom 10-30 min and is washed out with absolute Schaudinn’s fluid Abolte tanot 4 pt Subimatesln 2pats If necessary 1-5 per cent glacial acetic acid may be added. Its « common,2 > 6 [A hpe re grogeeohiockdccghigenipg-PMEElpg-uHE8dy Google eromosomes sharma sama Books QQ BB) co Asttomunry ~ —vtereview If necessary I-S per cent glacial aveti acid may be added. I fixative for Protozoa and is also used for higher organisms. I Washing, [Except in special cases, the tissue is thoroughly washed to remove all races I of fixing chemical Different periods of washing in running wate, from 1 hto ‘overnight, are employed, depending on the nature and the thickness of the tissue and the fixing fuid used. ‘Comparatively hard tissue, like lower-buds, root tips or bulk masses of ‘animal tissue, are put into perforated corked porcelain thimbles under running water. Ifthe tissue is very small or delicate, it ‘original tube, The fixing fluid is drained off and the tissue is washed in successive changes of warm (44°C) water at half-hourly intervals. “Alcoholic fxatives should be washed out with alcohols of approximately ‘the same percentage as that of the original solution, Reagents containing picrie acid should always be washed out with ethanol and never with water unless there is another constituent present inthe fluid which fixes chromatia 1€ > © pe lake goge ranch -ctagiaciAlsyn-PMcepg OM PERG Google ctromosames sharma sharma Books QQ BB) © Aattomanray » —_vereview ren << > © 2 Processing In some cases, however, log washing in water is unnscessey. Randolph 1 (0935) and later Upcot and La Cour (936) omitted washing altogether in ‘studying root tip chromosomes. They transferred the root tips directly from the heave to the deyerating agent, 70 pr cnt ethanol Dehydration Since the embedding material is usually immiscible with water and aqueous solutions, itis necessary to dehydrate the tissues before they can be em- becided, Tissues should not be transferred directly from water or aqueous solution to the undiluted dehydrating agent because an unequal shrinkage land distortion of the tissue are caused. The tissue should be passed through a ‘series of solutions, each containing a mixture of the dehydrating agent and a 1€ > © peak googacanickr-ctagiaciAlsyn-PMcepg-oMPERdy-h lee TOSan de QS Google —crromosomes sharma sharma Books ae 22 Aadtomyltray * —Witervew pons <> By ise both dehydration and hardening effects, The tisse is generally, ‘passed through successive grades containing 30, 0, 70, 80, 90, 95 per cent and absolute ethan, the period of treatment being variable, depending upon I ‘the nature and thickness of the specimen. For tissue ofthe size generally used {for chromosome study, | hin each is quite long enough, while for plant tissue, ‘overnight treatment in each of 70 per cent and absoluic ethanol is found to bbe most effective, The tissue ean be stored in 70 percent ethanol, if necessary ‘The different ethanol grades can be prepared from rectified spirit (approx. {96 per cent ethanol) as itis cheaper than absolute ethanol; Table 4.1 gives the ‘lative proportions of resid spirit and distilled water required to prepare ifferent concentrations, ‘Variations of the ethanol-dehydration schedule can be made, depending ‘on the nature ofthe tssue. The entire process can be speeded up for thin and “Table 44 Proparation of ethan of required conentration using rte spi approe 96 per set)Bi iitpe /hooka google coin ocks" ay SAAN EN PERI Patdomorrs arsenate OHARA Google —erromosomes sharma sharma EE : Books QQ BB) © Aattomanray » —_vereview ren << > © “able 41 Preparation of ean of requed concentration using rtd spit Gpproe v6 perce) Votan of eed pir Votan of war “Eka! concnraton ion wow hain we us i seasee delicate vssue, and some workers, instead of passing the tissue through different grades, prefer to increase the concentration of the medium contain- ing te tissue gradually by adding drops of strong ethanol at fixed intervals “The disadvantages of using ethanol as a denydrating agent ure: (a) the