What is glioblastoma?

Glioblastoma is a highly aggressive brain tumor characterized by rapid growth and infiltration into surrounding brain tissue.

How does confocal microscopy work?

Confocal microscopy uses laser light to illuminate fluorescently labeled samples, capturing images at various depths to create 3D representations.

What is the significance of using fluorescent proteins?

Fluorescent proteins allow for the visualization of specific cells or cellular processes in real-time, facilitating dynamic studies of cell behavior.

Why is tracking cell migration important?

Tracking cell migration is crucial for understanding tumor progression and developing effective treatments for cancer.

What are Z-stack images?

Z-stack images are a series of images taken at different depths within a sample, which can be combined to create a 3D view of the structure.

What role does temperature control play in cell culture?

Maintaining temperature control is essential for cell survival and function during imaging experiments.

Imaging Tumor Cell Migration in Glioblastoma Tissue Using Time-Lapse Scanning Confocal Microscopy

JoVE Encyclopedia of Experiments

Neuroscience

0 views • 2:07 min • June 17th, 2025

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Begin with a glioblastoma tissue slice obtained from a surgically excised tumor.

Maintain the tissue slice in a culture medium to support cell survival.

Tumor cells within the slice express a green fluorescent protein, or GFP, which facilitates the visualization of tumor cells during cell migration.

Place the culture plate containing the tissue slices on the stage of the confocal microscope.

Position a stage-top incubator to maintain the temperature of the medium.

Select an objective and visualize the slice cultures under the microscope.

Microscopes use laser light to excite the fluorescent proteins in the cells.

Now, Scan the sample layer by layer to capture images at various depths.

Stack the images to create a detailed 3D representation of the cells.

Then, capture Z-stack images at regular intervals to track cell movement and direction, enabling the study of tumor cell migration over time.

Place the dish into a 37 degree, 5% CO₂ sealed microscope stage top incubator of a confocal microscope. Use a long working distance, 10x air objective in conjunction with single or multi-photon imaging to capture three dimensional, time-resolved images of cell migration. Visualize the slice with the microscope, locating a suitable field with an adequate density of fluorescently labeled tumor cells between the slice edge and the center of the tissue. Then in the multidimensional analysis mode of the imaging software, set the center of the z-stack such that all of the positions to be imaged contain a visible fluorescent cellular signal. And set an adequate time for each z-stack acquisition.