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Simple Elimination of Background Fluorescence in Formalin-Fixed Human Brain Tissue for Immunofluorescence Microscopy
JoVE Journal
Biochemistry
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JoVE Journal Biochemistry
Simple Elimination of Background Fluorescence in Formalin-Fixed Human Brain Tissue for Immunofluorescence Microscopy
DOI:

16:31 min

September 03, 2017

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Chapters

  • 00:12Title
  • 00:48Construction of photobleaching aparatus and solutions
  • 02:30Photobleaching pre-treatment of tissue
  • 03:37Immunofluorescence
  • 09:35Fluorescence microscopy
  • 12:13Representative results
  • 15:58Conclusion

Summary

Automatic Translation

Background autofluorescence of biological samples often complicates fluorescence-based imaging techniques, especially in aged human postmitotic tissues. This protocol describes how autofluorescence from these samples can be effectively removed using a commercially available light emitting diode light source to photobleach the sample prior to immunostaining.

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