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slide preparation for electron microscope by hridoy Krishna Borah.pptx

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Hridoy Krishna Borah

1) The document discusses the process of slide preparation for electron microscopy. This includes cleaning, fixation, dehydration, infiltration, embedding, and sectioning of samples. 2) Key steps include fixation to preserve the sample using chemicals like glutaraldehyde or osmium tetroxide, dehydration using a graded ethanol series, and infiltration and embedding in epoxy resin. 3) The prepared blocks are then sectioned very thinly and viewed under the electron microscope. Other techniques mentioned include cryofixation, negative staining, and shadow casting. Proper sample preparation is important for high quality electron microscopy.

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SLIDE PREPARATION FOR ELECTRON MICROSCOPE SUBMITTED BY HRIDOY KRISHNA BORAH BSC 5th SEM RoLL NO-116
Introduction History 際際滷 preparation for electron microscope Other methods use for slide preparation Conclusion
INTRODUCTION Electron microscope are scientific instrument that use a beam of energetic electron to examine objects on very fine scale. An electron microscope can magnify between 1 and 50 million times depending on which type you use!
History Transmission electron microscope (TEM) was the first type of electron microscope to be developed. Developed by Max Knoll and Ernest Ruska in 1931. Max knoll Ernest Ruska
際際滷 preparation for electron microscope Sample preparation is important for electron microscopy.There are three main steps of sample preparation 1.Processing 2.Embedding 3.Polymerizing
Processing Its includes cleaning, fixation,rinsing,post fixation, dehydration, infiltration. a)Cleaning The proper cleaning of surface is very important. 0.1M cocodylic acid buffer( pH 7.3 )use as cleaner at normal room temperature . b) Fixation This is done to preserve the sample and to prevent further the deterioration. Eg-Gluteraldehyde fixation for Protein
c) Rinsing The should be washed with a buffer to maintain the pH. d)Post fixation A secondary fixation is done to increase stability and contrast the fine structure. Normally it is done with Osium tetroxide(OsO4)
e) Dehydration The specimen can be dehydrated in grade series of ethanol. f) Infiltration Epoxy resin is use to infiltrate the cell .
Embedding After the Processing the next Step is embedding . This is done Using fold mold . POLYMERIZING Next is Polymerizing step in which the resin is allowed to set overnight at a temperature of 60 degree at in a oven.
Sectioning The specimen must be cut into very thin section for electron microscopy so that the electron are semi-transparent to electron.
Other methods of slide preparation 1.Crytofixation- Like Chemical fixative ,its stop the Metabolic process and preserve the biological structure. 2.Negative staining the main purpose of it to surround or embed the biological object in a suitable electron dense material. 3.Shadow casting The grid containing the specimen are placed in sealed chamber which is evacuated by vaccum pump.
4.Freeze fracture replication and freeze etching Small pieces of tissues are place in metal disk and rapidly frozen
Conclusion Most of chemical use in the procedure are dangerous and hazardous. These procedure are time consuming and required skills
References Wikipedia Google images Research Gate
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slide preparation for electron microscope by hridoy Krishna Borah.pptx

  • 1. SLIDE PREPARATION FOR ELECTRON MICROSCOPE SUBMITTED BY HRIDOY KRISHNA BORAH BSC 5th SEM RoLL NO-116
  • 2. Introduction History 際際滷 preparation for electron microscope Other methods use for slide preparation Conclusion
  • 3. INTRODUCTION Electron microscope are scientific instrument that use a beam of energetic electron to examine objects on very fine scale. An electron microscope can magnify between 1 and 50 million times depending on which type you use!
  • 4. History Transmission electron microscope (TEM) was the first type of electron microscope to be developed. Developed by Max Knoll and Ernest Ruska in 1931. Max knoll Ernest Ruska
  • 5. 際際滷 preparation for electron microscope Sample preparation is important for electron microscopy.There are three main steps of sample preparation 1.Processing 2.Embedding 3.Polymerizing
  • 6. Processing Its includes cleaning, fixation,rinsing,post fixation, dehydration, infiltration. a)Cleaning The proper cleaning of surface is very important. 0.1M cocodylic acid buffer( pH 7.3 )use as cleaner at normal room temperature . b) Fixation This is done to preserve the sample and to prevent further the deterioration. Eg-Gluteraldehyde fixation for Protein
  • 7. c) Rinsing The should be washed with a buffer to maintain the pH. d)Post fixation A secondary fixation is done to increase stability and contrast the fine structure. Normally it is done with Osium tetroxide(OsO4)
  • 8. e) Dehydration The specimen can be dehydrated in grade series of ethanol. f) Infiltration Epoxy resin is use to infiltrate the cell .
  • 9. Embedding After the Processing the next Step is embedding . This is done Using fold mold . POLYMERIZING Next is Polymerizing step in which the resin is allowed to set overnight at a temperature of 60 degree at in a oven.
  • 10. Sectioning The specimen must be cut into very thin section for electron microscopy so that the electron are semi-transparent to electron.
  • 11. Other methods of slide preparation 1.Crytofixation- Like Chemical fixative ,its stop the Metabolic process and preserve the biological structure. 2.Negative staining the main purpose of it to surround or embed the biological object in a suitable electron dense material. 3.Shadow casting The grid containing the specimen are placed in sealed chamber which is evacuated by vaccum pump.
  • 12. 4.Freeze fracture replication and freeze etching Small pieces of tissues are place in metal disk and rapidly frozen
  • 13. Conclusion Most of chemical use in the procedure are dangerous and hazardous. These procedure are time consuming and required skills
  • 14. References Wikipedia Google images Research Gate

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