Microscopy report outline

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20.309: Biological Instrumentation and Measurement

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General guidelines

  • Submit one paper per group.
  • Bulleted list or outline format is encouraged
  • Report numerical results properly
    • Indicate the units of measurement
    • Include the sample size and an appropriate measure of variability, such as a range, standard deviation, or standard error
    • Use the abbreviation "s.d." for standard deviation and "s.e.m." for standard error after the "±"
    • For example: 1.21 ± 0.03 GW (±s.d., n=42)
  • Explain how you analyzed the data
    • Summarize the algorithm used for all calculations and analyses
    • Provide an outline of each MATLAB (or other language) function or script you used
    • Put the complete code in an appendix
    • Indicate the source of any code you did not write yourself
  • Discuss your results
    • Compare your results to theoretical predictions, reported values from literature, or other students' results
    • Explain any factors that may have affected your results
    • Describe what you would do differently if you had the opportunity to do the experiment again.
  • Provide a detailed, correct, and comprehensive discussion of error sources for each measurement
    • Is the resulting error random or systematic?
    • If the error is systematic, can you determine the sign of the effect and its approximate magnitude?
    • How could the error be reduced?
  • Present data properly
    • Images should include a scale bar
    • Plots should have a clear title; axes should be labeled with units; use a plot legend when appropriate

Part 1: Microscope construction and bright field characterization

Microscope documentation and design

  • Microscope block diagram, including all optical elements and relevant distances. It is unnecessary to document the details of the mechanical construction.
  • Design calculations and considerations
  • Photograph of your setup (optional, but nice)

Microscope characterization

  • Magnification and field of view
    • Include a table with the following values for the 10X, 40X, and 100X objectives:
      • Theoretical resolution
      • Actual magnification by multiple measures (Air Force Target, Ronchi Ruling, microspheres)1
      • Actual field of view (FOV)1
    • Example transilluminated pictures from each objective
  • Measured resolution of 40X (or 100X) objective1
    • Images used for resolution estimation (preferably with fit – see plotgaussfit command)
    • Estimate of FWHM by Gaussian fitting1
    • Bullet point outline of data analysis methodology
    • Comments on estimated versus theoretical value
  • Stability
    • X-Y plots of sum and difference tracks for fixed particles.
    • MSD versus time interval for sum and difference tracks1
    • Bullet point outline of data analysis methodology
    • Comments on observed vs. expected data trend

Part 2: Fluorescence microscopy

  • Unprocessed Images of fluorescent bead associated reference images
  • Corrected images
  • Image or surface plot (see surf command in Matlab) of correction applied
  • Histograms of original and corrected images
  • Bullet point outline of image processing methodology

Parts 3+: Particle tracking and other experiments

  • X-Y plots of tracks for all samples
  • MSD versus time interval plots for measured samples
  • Estimate of diffusion coefficient, viscosity and other mechanical properties for each sample
  • Comments on results, specifically microscope stability, resolution,
  • Comment extensively on sources of error and approaches to minimize them, both utilized and proposed
  • Bullet point outline of all calculation and data processing steps


1Remember to include uncertainty and a discussion of error sources for all numerical results.