This study demonstrates a method for visualizing glioblastoma cell migration using confocal microscopy. By employing tissue slices from excised tumors, researchers can track the movement of fluorescently labeled tumor cells over time.
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.
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