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Fat Body Staining for Gut

Authors: Catherine Lian and Brian Hodge, updated date: , view: 254, Q&A: 0
Tags: Fly and Drosophila


  1. Fat bodies are dissected in Schneider’s drosophila medium (1×);

  2. Fixed in 4% paraformaldehyde (PFA) for 45 min at room temperature (ask Mark).

  3. Samples are rinsed, washed with 0.1% Triton X-100 in 1× PBS (1× PBT) (3 times, 10 min/time).

  4. Blocked in 5% horse serum in 1× PBT at 4°C overnight.

  5. Incubate primary antibodies (Rat-anti-TIM, 1:500; Guinea pig-anti-PER, 1:500) at 4°C overnight.

  6. Washed with 1× PBT for 1 hour.

  7. Secondary antibodies (Rat-Alexa 488, 1:500; Guinea pig-Alexa 555, 1:500) were incubated at 4°C overnight.

  8. Washed with 1× PBT for 1 hour.

  9. Stain for 15 min with DAPI (Sigma; 0.1 μg/ml)

  10. Remove DAPI and add PBT.

  11. The samples were mounted in mounting medium mowiol (ask Mark).

  12. All images were captured by a Zeiss LSM780 inverted confocal microscope and were processed in Adobe Photoshop and Illustrator.

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