What are rigor and reproducibility?
Rigor means following procedures that will increase the likelihood of obtaining an accurate representation of the phenomenon under study.
Reproducibility means recording and communicating those procedures such that they can be replicated accurately.
If your experiments are performed rigorously and reproducibly, other researchers should be able to replicate your procedures, have a high likelihood of obtaining similar results on similar samples, and have those results be an accurate representation of the phenomenon being studied.
Rigor requires you pay attention to:
- The reagents and protocols used to prepare your samples. MSL staff follow best practices in electron microscopy sample preparation, and can provide the exact protocols that were followed for inclusion in methods sections of papers.
- The condition of the instruments used to take images and analyze images. MSL performs weekly maintenance on our instruments; the specific checks we perform are explained here.
- The controls included in your experiments. MSL provides general recommendations for controls in light-microscopy sample preparation.
- The way in which you decide which parts of your sample to study. This can involve decisions on which parts of samples to prepare (for example, how to section a piece of tissue, which sections to stain) and where in those samples to take images. MSL provides suggestions for how to make sampling decisions, as well as resources for further reading.
- The way in which you decide how many images to take per sample, as well as how many samples to image. This mainly involves power analysis, a statistical technique used to determine how many samples are needed under different experimental circumstances. MSL can provide resources for researchers to do this properly.
- The way in which a microscope is set up for imaging. MSL provides individual, detailed training with each researcher’s samples to ensure they know how to properly set up imaging conditions.
Reproducibility requires you to pay attention to:
- Documenting parameters used for image acquisition and saving data in proper formats. MSL provides advice on how to accurately record those parameters, as well as which formats are appropriate when saving data during our individual training sessions.
- Documenting procedures used for imaging and image analysis. MSL can provide advice on what constitutes a proper record of those procedures. Details here.
- Properly writing methods sections. MSL can provide detailed hardware information as well as guidelines for properly writing methods sections of papers. Details of how to retain metadata here, and how to write methods sections here.