Ali Shaawiat
I think this site will help you
http://www.microscope-microscope.org/basic/preparing-microscope-slides.htm
2 votes
0 thanks
Heba Turky
I think this site will help youhttp://www.microscope-microscope.org/basic/preparing-microscope-slides.htm
2 votes
1 thanks
Khalil Chelab
Dear Dr. Atiaf:
These important links for you:
http://www.made-in-china.com/cs/hot-china-products/Microscope_Slides.html?gclid=COKxtpfbs9MCFcJAGwod824CHA
http://www.microscopemaster.com/microscope-slides.html
2 votes
2 thanks
Qassim Kshash
Dear, slide preparation for microbiology, histology ,hematology ,please limit
2 votes
2 thanks
Zeena Fouad
www.ivyroses.com/HumanBody/Histology/How-to-Prepare-Histology-Slides.php
2 votes
1 thanks
Furkan Alaraji
TISSUE PREPARATION • The most widely used method of studying tissues is using histological slides. • The tissue in the slide must reflect the actual nature of the tissue in the body • To insure that, tissues to be studied must pass through a series of steps before examination • These steps are in sequential order • Fixation is a complex series of chemical events that differ for the different groups of substance found in tissues. TISSUE FIXATION Aim of Fixation: 1- To prevent autolysis and bacterial attack. 2- To fix the tissues so they will not change their volume and shape during processing. 3- To prepare tissue and leave it in a condition which allow clear staining of sections. 4- To leave tissue as close as their living state as possible, and no small molecules should be lost. •Fixation is a reaction between the fixative and proteins in the specimen which form a gel, so keeping every thing as their in vivo relation to each Types of Fixative • Acetic acid • Formaldehyde 10% • Ethanol • Glutaraldehyde • Methanol • Picric acid • Osmic acid (Osmium tetroxide) 6. TISSUE PROCESSING • Aim: Is to embed the tissue in a solid medium firm enough to support the tissue and give it sufficient rigidity to enable thin sections to be cut , and yet soft enough not to damage the knife or tissue. • Stages of processing: 1.Dehydration. 2- Clearing. 3- Embedding. Dehydration • To remove fixative and water from the tissue and replace them with dehydrating fluid. • Delicate specimens are dehydrated in a graded ethanol series from water through 10%-20%-50%-95%-100% ethanol to minimize tissue distortion from diffusion currents. • In the paraffin method, dehydration from aqueous fixatives is usually initiated in 60%- 70% ethanol, progressing through 90%-95% ethanol, absolute ethanol before proceeding to the clearing stage. Types of dehydrating agents • Ethanol • Methanol • Acetone • Tissues may be held and stored indefinitely in 70% ethanol without harm Clearing • Replacing the dehydrating fluid with a fluid that is totally miscible with both the dehydrating fluid and the embedding medium. • Choice of a clearing agent depends upon many factors Types of Clearing Agents • Xylene. • Toluene. • Chloroform. • Benzene. • Propylene oxide Embedding • Is the process by which tissues are surrounded by a medium such as agar, gelatin, or wax which when solidified will provide sufficient external support during sectioning. • A substances added alone or in combination to the wax to: Improve ribboning. Increase hardness. Decrease melting point Improve adhesion between specimen and wax Embedding Moulds A. paper boat mould B. metal boat mould C. Dimmock embedding mould D. Peel-a-way disposable mould E. Base mould used with embedding ring ( F) or cassette bases (G) CUTTING • using the microtome • A microtome is a mechanical instrument used to cut biological specimens into very thin sections for microscopic examination. • Most microtomes use a steel blade and are used to prepare sections of animal or plant tissues for histology. Microtome knives • STEEL KNIVES • NON-CORROSIVE KNIVES FOR CRYOSTATS • DISPOSABLE METAL BLADES • GLASS KNIVES • DIAMOND KNIVES STAINING Hematoxylin and Eosin (H & E) • H & E is a charge-based, general purpose stain. • Hematoxylin stains acidic molecules shades of blue. • Eosin stains basic materials shades of red, pink and orange. • H & E stains are universally used for routine histological examination of tissue sections. Staining Procedure • Deparaffinize and hydrate to water • If sections are Zenker-fixed, remove the mercuric chloride crystals with iodine and clear with sodium thiosulphate (hypo) • Mayer's hematoxylin for 15 minutes • Counterstain with eosin from 15 seconds to 2 minutes depending on the age of the eosin, and the depth of the counterstain desired • Dehydrate in 95% and absolute alcohols, two changes of 2 minutes each or until excess eosin is removed • Clear in xylene, two changes of 2 minutes each • Mount in Permount or Histoclad Staining machine • Basic dyes stain: Heterochromatin Nucleic acids Ribosomes Cartilage • Acidic dyes stain: Filaments Mitochondria Collagen Muscle fibers Additional Dyes • Many tissue components can not be stained with (Hematoxylene and Eosin). • Other dyes are used to specifically stain certain tissue components Resorcin-Fuchsin for elastic fibers Silver stain for reticular fibers and basement membrane Periodic-Acid Schiff (PAS) Reaction for CHO. REGARDS
Hi
thanks for revealing question. i will explain the method with details
2 votes
0 thanks
Furkan Alaraji
TISSUE PREPARATION • The most widely used method of studying tissues is using histological slides. • The tissue in the slide must reflect the actual nature of the tissue in the body • To insure that, tissues to be studied must pass through a series of steps before examination • These steps are in sequential order • Fixation is a complex series of chemical events that differ for the different groups of substance found in tissues. TISSUE FIXATION Aim of Fixation: 1- To prevent autolysis and bacterial attack. 2- To fix the tissues so they will not change their volume and shape during processing. 3- To prepare tissue and leave it in a condition which allow clear staining of sections. 4- To leave tissue as close as their living state as possible, and no small molecules should be lost. •Fixation is a reaction between the fixative and proteins in the specimen which form a gel, so keeping every thing as their in vivo relation to each Types of Fixative • Acetic acid • Formaldehyde 10% • Ethanol • Glutaraldehyde • Methanol • Picric acid • Osmic acid (Osmium tetroxide) 6. TISSUE PROCESSING • Aim: Is to embed the tissue in a solid medium firm enough to support the tissue and give it sufficient rigidity to enable thin sections to be cut , and yet soft enough not to damage the knife or tissue. • Stages of processing: 1.Dehydration. 2- Clearing. 3- Embedding. Dehydration • To remove fixative and water from the tissue and replace them with dehydrating fluid. • Delicate specimens are dehydrated in a graded ethanol series from water through 10%-20%-50%-95%-100% ethanol to minimize tissue distortion from diffusion currents. • In the paraffin method, dehydration from aqueous fixatives is usually initiated in 60%- 70% ethanol, progressing through 90%-95% ethanol, absolute ethanol before proceeding to the clearing stage. Types of dehydrating agents • Ethanol • Methanol • Acetone • Tissues may be held and stored indefinitely in 70% ethanol without harm Clearing • Replacing the dehydrating fluid with a fluid that is totally miscible with both the dehydrating fluid and the embedding medium. • Choice of a clearing agent depends upon many factors Types of Clearing Agents • Xylene. • Toluene. • Chloroform. • Benzene. • Propylene oxide Embedding • Is the process by which tissues are surrounded by a medium such as agar, gelatin, or wax which when solidified will provide sufficient external support during sectioning. • A substances added alone or in combination to the wax to: Improve ribboning. Increase hardness. Decrease melting point Improve adhesion between specimen and wax Embedding Moulds A. paper boat mould B. metal boat mould C. Dimmock embedding mould D. Peel-a-way disposable mould E. Base mould used with embedding ring ( F) or cassette bases (G) CUTTING • using the microtome • A microtome is a mechanical instrument used to cut biological specimens into very thin sections for microscopic…
Hello
Hithanks for revealing question. i will explain the method with details
3 votes
1 thanks
Hassan Hammadi
Dear Dr. Atiaf
I think this link will help you
http://www.microscope-microscope.org/basic/preparing-microscope-slides.htm
regards
2 votes
0 thanks
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