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.

1. SLIDE PREPARATION FOR
ELECTRON MICROSCOPE
SUBMITTED BY
HRIDOY KRISHNA BORAH
BSC 5th SEM
RoLL NO-116

2. • Introduction
• History
• Slide 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. Slide 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
• It’s 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

15. THANK YOU