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A quantitative investigations of plants expressing green fluorescent protein (GFP) using a home-made camera attachment

In this laboratory exercise students use digital cameras to measure surface fluorescence of transgenic plants expressing green fluorescence protein (GFP). Inquiry pedagogy is used to teach processes used in scientific investigations and how GFP is used in reporter gene systems. Students start by making observations involving induction of expression of a GFP expressing reporter gene and then use their observations to develop a testable hypothesis regarding possible environmental or developmental signals responsible for inducing gene expression. The students design and conduct an experiment to evaluate their hypothesis. In their experiments, the students assess GFP expression by taking photographs with a digital camera attached to an epifluorescence filter cube. Open source software is used to quantify the amount of green surface fluorescence observed in the students’ photographs. After performing basic statistical analysis, the students report on their findings by writing a scientific poster.

The epifluorescence filter cube that enables the camera to detect GFP fluorescence is relatively inexpensive because it is constructed largely from consumer goods. The filter holder is made from hardboard. The illumination source is a 5 watt light emitting diode (LED), and the lens tube is made from plastic plumbing pipe. The specialty materials are a set of dichroic filters. To reduce costs, mass production machine vision filters are used as excitation and barrier filters. The most expensive component is the beam splitting filter.

Materials included with the manuscript are instructor’s notes, instructions on how to construct the camera attachment, a student handout explaining how to use ImageJ to quantify surface fluorescence, an example assignment where students investigate the specificity of the Arabidopsis thaliana alcohol dehydrogenase (ADH) gene promoter, Power Point slides with a lecture outline, a grading rubric, and an example of a good scientific poster.
Format
Primary or BEN resource type
Secondary resource type
General Biology Core Concepts
Discipline Specific Core Concepts
Life science discipline (subject)
Keywords acclimation, green fluorescent protein, reporter gene, scientific method
Key taxa Arabidopsis thaliana
Audience
Intended End User Role
Language
Educational Language
Pedagogical Use Description The exercise is an inquiry investigation of an environmentally regulated promoter controlling the expression of the green fluorescent protein reporter gene. The students conduct experiments of their own design to identify environmental factors that control gene expression. The students use basic statistics to evaluate their results.
Primary Author Controlled Name
Primary Author Affiliation University of Detroit Mercy
Primary Author email bakerss@udmercy.edu
Submitter Name Stokes S Baker
Submitter Email bakerss@udmercy.edu
Rights All materials and images are original. The author owns the copyrights.
Date Of Record Submission 2016-01-05

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