Image Vet 70 Plus Manual

Image Vet 70 Plus Manual



 
 
 
 
 
 
 

Image Vet 70 Plus Manual

As the acquisition of the control images can be time consuming, it is important to start by acquiring a house keeping image of the control field to serve as a reference. When measuring the fluorescent or brightfield signal in an experiment, it is often important to know the intensity of the control field. Thus, on the fluidic stage plate, the area that is to be imaged following the acquisition of the tissue should be located in the center of the field and be at the same position throughout the experiment. Thus, it is critical that the acquisition of the control reference image is performed at the same position as the other fields and with the same setup that will be used for the image of interest.

A simple and obvious control is to include a live cell monolayer on a cover slip. When growing cells directly on the cell monolayer on the cover slip, the cultures often are exposed to fluctuations in nutrient and waste levels and lack of growth media can affect the cellular signals within the cultured tissue over time. Therefore, the correct acquisition of the control image and subsequent identification of cellular features should be performed as soon as possible after the cultured tissue has been delivered to the fluidic stage plate for image capture with the LED and camera setup used for the experiment. The timing of the acquisition of the control image can be critical. For culture conditions where the duration of the experiment is known, it can be acquired immediately following loading of the tissue but if the experiment has a long duration, it may be preferable to have a control image acquired every few hours to make sure that the images are uniform and the tissue is reproducible.

In this study, the control images were acquired using the process of dual fluorescent staining as the images were developed and captured during the live acquisition of the tissue for the harvested images. A key example of performance is the ability to detect early changes in cellular fluorescence that may occur during image acquisition. To maintain the tissue viability by minimizing the duration of time it is exposed to the fluidic stage, moderate incubation of the tissue with Cell Stain that has a profile matched to the tissue at the lowest possible amount needed for the desired signal is preferred. With this technique, the sample will be moderately stained over multiple alternating cycles (approximately two to four rounds of alternating light and dark stain), which will allow for changes in cellular uptake of the markers prior to the ultimate image capture.

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