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Immunofluorescence Staining

Direct immunofluorescence method is used to detect the deposit of immunoglobulins, complement components, fibrinogen, etc. in tissues. This technique is usually performed on frozen sections. The primary antibody is conjugated to fluorescein binds directly with the antigen and can be detected by the fluorescent tag using a fluorescent microscope.


Frozen tissue only.

  1. Fresh tissue (kidney, lung, skin, etc.) is frozen in -80°C freezer.
  2. Embedding: OCT medium sectioning at 7μm.


  1. Use a frozen control of tonsils.


  1. Place some OCT in plastic freezing boat.
  2. Place tissue in OCT and orient according to standard embedding protocol.
  3. Freeze in -80° Freezer.
  4. Remove block from freezer and place in cryostat.
  5. Cut sections from frozen block and place on coated slides. Sections should be cut at 7 microns on the cryostat.
  6. Fix in cold acetone for 10 minutes.
  7. Air dry.
  8. Place slides in washing PBS for 5 minutes. OCT medium will dissolve in this wash.
  9. Place antibodies on slides for 30 minutes. Slides must be kept in dark.
  10. Rinse in PBS.
  11. Coverslip with fluorescent mounting medium.
  12. Store slides in cardboard folder in refrigerator until ready to screen.

Note: Using the Ventana automated procedure, slides will be placed on machine at Step 9 and resumed with Step. 10.

2016 EDRN PI Orientation

The New and Continuing EDRN Principal Investigator Orientation will take place March 14–15, 2016 in Bethesda, Maryland.

Announcement 11/05/2015

Thank you to everyone who made the 29th EDRN Steering Committee Meeting a success. The orientation for new and continuing EDRN PIs will be held Monday-Tuesday, March 14-15, 2016, on the NCI campus. More information will be available later this autumn.

Announcement 06/30/2015

A funding opportunity for a new pancreatic cancer initiative, called The Pancreatic Cancer Detection Consortium (U01), has been released. For more information, please go to /grants/guide/ pa-files/ PAR-15-289.html.