Introduction fluorescence microscopy

 Microscopy in general

Resources on microscopy:

Fluorescent spectra tools:

Considerations before performing Microscopy data acquisition

  • Choose the right technique for your experiment (JCS 2007 (122) 753)
  • Have a plan before acquiring images.
  • Choose proper materials:

coverslip thickness (#1.5), live cell chambers, fixation buffers, dyes, affinity labeling tools, mounting media.

  • Test labelling strategies thoroughly before performing large scale sample sets or intravital microscopy.

nuclear counterstain, cell mask, specific stain, multi dye stain, live cell timing, in vivo tile/stack size, fiducial marker etc.

  • Consider that data analyses often require labeling of defined structures to which you can relate your stain of interest.
  • Rethink your experiment to implement a workflow from doing the experiment, imaging it and analyzing the data. All three aspects will take time and effort to complete.
  • Estimate the resolution (spatially, temporally, spectrally) you need to draw a conclusion
  • Estimate the optimal image data volume you need to draw a conclusion (consider population variation of a parameter and group size to reliably measure changes).