ELECTRON MICROSCOPY FOR PATHOLOGISTS

Training: Darkroom Procedures & Digital Imaging

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Training Menu 
Introduction to the Resident Rotation    
Learning Objectives 
About the TEM
Tissue Preparation
Alternative Tissue Preparation from Paraffin Block
Ultramicrotomy
Darkroom Procedures and Digital Imaging
List of Required Cases 

Darkroom Procedures

After photographs are taken in the EM, follow these steps:

1. Develop the negative:

Step 1 - developer		2-4 minutes
Step 2 - fixer		5 minutes
Step 3 - perma wash		2 minutes
Step 4 - photo flo		5 washes
Step 5 - tap water wash

2. Dry the negative in the oven at 60°-70°C for 15-20 minutes.
3. Identify envelopes containing negatives with a negative number and an EM number.
4. Print the paper microphotograph:

• Step 1--put the negative in the enlarger and expose the paper
• Step 2--develop the paper with the automatic processor (Rapidoprint DD370 AGFA). Processor steps are:
  1. activator bath
  2. stabilizing bath
  3. tap water baths (2)
  4. print drying

5. For identification, label the back of the microphotographs with a negative number, an EM number, a surgical pathology number, and indicate final magnification.

Digital Imaging
Negatives are scanned with a high-resolution scanner and the image is converted to positive. After the images are captured they are stored in the pathology server and are readily available for diagnosis, presentations, or teaching purposes from any PC.

Alternatively, the scanned images also can be obtained from paper printed microphotographs.

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