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Botany JoVE (Open Access) Citable · DOI

Robotics and Dynamic Image Analysis for Studies of Gene Expression in Plant Tissues

DOI: 10.3791/1733-v
What you'll learn
  • ✓ Perform particle bombardment for GFP gene introduction into plant cells
  • ✓ Operate robotic systems for semi-continuous automated image collection
  • ✓ Analyze time-series fluorescence images using ImageJ and ImageReady
Protocol

We report a method for introduction, tracking and quantitative analysis of GFP expression in plant cells. This method utilizes a custom-designed robotics system for semi-continuous image collection from large numbers of samples, over time. We also demonstrate the use of ImageJ and ImageReady for analysis of image series.

Difficulty
advanced
Total time
~2-4 weeks (including growth period, gene introduction, and image acquisition timeline)
Model organism
Plant tissues (species not specified in abstract)
Biosafety
BSL-1

Steps

1
Review gene expression tracking methodology and objectives

Understand the workflow for introducing, tracking, and quantifying GFP expression in plant cells using robotic and imaging approaches.

▶ 00:45
2
Introduce genes via particle bombardment

Apply particle bombardment technique to deliver GFP genes into plant tissue samples.

▶ 01:34
3
Collect automated serial images with robotic system

Operate custom-designed robotics to perform semi-continuous image acquisition from multiple samples over extended time periods.

▶ 04:51
4
Analyze image series using ImageJ and ImageReady

Process and quantitatively analyze collected image sequences to measure GFP expression patterns in plant cells.

▶ 07:44
5
Interpret representative results and expression patterns

Review and evaluate gene expression imaging outcomes from plant tissue samples.

▶ 09:14
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