NEU 3400 - Fluorescence Spectra and Filters Homework

• The last page of the study guide has an example fluorophore and several filter sets that could be used to visualize it.
• Generate a separate document (Word or written by hand) to explain which filter set is best, in your opinion.
• Be prepared to defend your answer, as well as your rationale for not selecting other filters, in class Thursday.
• You will turn in this page or document as homework on Thursday. See you then!

• Alexa Fluor 647

  • Bright , Far-Red–Fluorescent dye

    • Should Use the CY5 Selection on Microscope Filter Wheel ( yellow light --> excites red dye )

      

  • Max Excitation Wavelength = 650 nm
  • Max Emission Wavelength = 665 nm
  • Laser Lines = 595 , 633 , 635 , 647
  • https://docs.abcam.com/pdf/immunology/fluorochrome_guide.pdf
  • https://www.thermofisher.com/us/en/home/life-science/cell-analysis/fluorophores/alexa-fluor-647.html

https://www.semrock.com/1optical-filters-for-laser-based-fluorescence-microscopes.aspx

https://www.edmundoptics.com/knowledge-center/application-notes/optics/fluorophores-and-optical-filters-for-fluorescence-microscopy/

• Dashed Line = Excitation Curve of Alexa Fluor 647
• Red Shaded Region = Emission Curve of Alexa Fluor 647

Possible Excitation + Emission Filter Combinations

Using Preset Cy5 ( Alexa Fluor 647 ) Excitation and Emission Filter