Live-cell Imaging of Glioblastoma Cell Motility

Cells were plated on laminin-coated Bioptechs delta-T dishes. For the duration of video acquisition, cells were maintained in a sealed chamber at 37 °C and 5% CO₂. Phase-contrast images were taken at 10-min intervals for 90 min total. Images were acquired with the 10× objective of a Zeiss Axiovert 200M microscope equipped with an AxioCam HRm CCD camera (Zeiss). Motility was quantitated using the MtrackJ plugin (42 (link)) in ImageJ software (National Institutes of Health) as described previously (15 (link)). For live cell imaging and video microscopy of actin, lentiviral particles expressing eGFP-tagged Lifeact (23 (link)) were made and used to transduce glioblastoma cells. pLenti Lifeact-EGFP BlastR was a gift from Ghassan Mouneimne (Addgene plasmid # 84383).

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Lavictoire S.J., Jomaa D., Gont A., Jardine K., Cook D.P, & Lorimer I.A. (2021). Identification of Rac guanine nucleotide exchange factors promoting Lgl1 phosphorylation in glioblastoma. The Journal of Biological Chemistry, 297(5), 101172.

Publication 2021

delta-T dishes Axiovert 200M microscope AxioCam HRm CCD camera

Actin Cells Dishes Glioblastoma Laminin Microscope Motility Phase contrast Plasmid Video microscopy

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Variable analysis

independent variables

Laminin coating of Bioptechs delta-T dishes

dependent variables

Cell motility quantitated using MtrackJ plugin in ImageJ software

control variables

Cells maintained in a sealed chamber at 37 °C and 5% CO2 during video acquisition
Use of phase-contrast microscopy with 10× Zeiss Axiovert 200M objective and AxioCam HRm CCD camera

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