Sample Preparation

Dont’s

  • Use antibodies labelled with any PE or APC labels; not visible with the confocal…..

Do’s

  • For multiple stainings; When using antibodies produced in the same species, use IgG specific secondary antibodies. Available from Molecular probes (Invitrogen life technologies).
  • Look in the Molecular Probes Handbook
  • Use the Spectra Viewer or Leica FluoScout
  • Alexa fluor labelled secondary antibodies are excellent for confocal use. Also available at invitrogen life technologies.
  • Use the Alexa fluor Selection Guide
  • Molecular probes also has two inclusion media, Prolong Gold with and without DAPI. So no more nuclear staining to, no more using nailpolish (Prolong Gold hardens out overnight) and stable staining upto a month! Ideal for preparing confocal experiments in advance. Cat numbers with DAPI; P36931 and P36935, without DAPI P36930 and P36934.
  • Use Cover glass no. 1.5H (lowest tolerance in thickness) for example
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