“Live cell imaging”
Introduction Live cell imaging is based on time laps images and it has various applications in biomedical and biology sciences (figure1). It gives spatial and temporal information as well as information about cell functions through monitoring and measuring the cellular dynamic process in biological structures. Successful live cell imaging is challenging as the cells needs to be kept at a good condition while making sure of the high quality imaging. There are different types of microscopes that serve the live cell imaging purpose such as Phase contrast microscopy, Fluorescent microscopy and widefield microscopy. Reference: https://www.thermofisher.com/fi/en/home/life-science/cell-analysis/cell-analysis-learning-center/molecular-probes-school-of-fluorescence/sample-considerations/live-cell-imaging.html
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Laser and green fluorescence excite the specimen and it goes under the mirrors scanning in the device. The emitted light by the laser passes a dichroic mirror and it focused to the pinhole (figure 4). Then it is detected by the microscope's camera or photodetector. It should be noted that the sample is being scanned point by point and to get a complete image the computer must build up the image of one pixel at a time. The mirrors in a confocal microscope are controlled by a computer that can store and analyze the produced data. Figure 4. Confocal microscope
Advantages and disadvantages of confocal microscopy One of the benefits of the confocal microscopy is producing a series of thin sections of a sample and creating a high resolution as well as 3D images that are taken from horizontal and vertical angels with the possibility to control the depth of field. Blocking out of the focus fluorescence light using the pinhole makes this imaging method more accurate. Confocal imaging method can be time-consuming as it scans the sample point by point. Moreover, it tends to be expensive. Phototoxicity and Photobleaching are other drawbacks of this