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QuickID Targeted Image Acquisition

Increase imaging efficiency using QuickID Targeted Acquisition

Investigators performing image-based high content screening may face several technical challenges if they need to image their samples at high resolution, at multiple wavelengths, and over a range of z-planes. Such experiments often have lengthy acquisition times (between several hours and a couple of days), a vast need of image storage space (100s of GBs), and high computational power requirements to keep image analysis time managable (the fewer hours the better).

 

A simple way to minimize all of these challenges is to acquire only the fields-of-view on the slide or in the microplate that contain objects of interest. This technique can be especially useful when the object of interest is scarcely abundant in each sample, randomly distributed in space, or only occurs as a rare event in some of the microwells. This solution, when put into use with our high-content imaging systems and image analysis software, dramatically reduces the time-to-result as well as image storage requirements.

QuickID allows the acquisition of the entire sample at low magnification, finds objects of interest or rare events through a fast image analysis process, and then re-acquires only these wells or sites at high magnification with a user configured protocol that may include the relevant wavelengths, z-stacks, and time-points. Targeted acquisition is performed by the MetaXpress software and can be used with the ImageXpress Micro 4 and the ImageXpress Micro Confocal systems.

quickid-targeted-acquisition

QuickID targeted acquisition works by acquiring entire field-of-view of a sample at low magnification, then finding objects of interest or rare events through a fast image analysis process, and then re-acquiring these images at high magnification.

 

Acquire data 10X faster from 3D cell cultures

In the application note “Acquire data 10X faster from 3D spheroids cultured using Symphony® and VersaGel®” a light crosslinkable hydrogel was used as a matrix to grow 1-4 spheroids/well in a 3 dimensional space. After staining, QuickID was performed using a 2X objective to capture the entire well in one field-of view. Objects of interest were identified and their X and Y coordinates automatically referenced so the ImageXpress Micro system obtained only relevant images at 10X magnification in three wavelengths across multiple z planes. With QuickID, acquisition time was 10X faster than imaging all sites in every well at high magnification and this also reduces the required storage space by 20X.

 

3D cell cultures

 

QuickID was used to streamline spheroid image acquisition. An image acquired at low magnification to view the entire well in one field-of-view was used to identify objects for automatic re-imaging at higher magnification using three wavelengths across multiple z planes.

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Other applications for targeted acquisition

QuickID streamlines the process so only relevant images are acquired which results in significant time-savings and reduced storage needs. Examples of other application areas that would benefit from the use of QuickID include live imaging of dividing cells or model organisms, and imaging specific areas of tissue sections on a slide.

 

  • Mitotic spindle

    Mitotic spindle

    In a normal cell population, only about 1-5% of cells are in mitosis. Targeted acquisition can be used to identify these cells in the whole population and then follow them over time in high magnification. Tracking and re-centering of single cells is possible should they move out of region during the acquisition time course.

    3D micro-spheroids

    3D micro-spheroids

    Micro-spheroids often accumulate at the edge of flat bottom plates. An initial transmitted light acquisition followed by a targeted z-stack acquisition of fluorescent makers can be performed to greatly reduce the number of images required.

  • Hippocampus in a brain slide

    Hippocampus in a brain slide

    Depending on the sample morphology and its preparation, the hippocampus and other parts of the brain could end up in various area on a slide. Targeted z-stack acquisition can be performed at multiple wavelengths for the study of co-localization.

    Heart beat in zebrafish

    Heart beat in zebrafish

    The position and orientation of a zebrafish in a well can vary. High magnification z-stack or time-lapse acquisitions of the heart, eye, brain, yolk sac, irridophores, or blood vessels can be performed using targeted acquisition.

  • Micronuclei

    Micronuclei

    At low magnification, it is difficult to accurately determine if an object is really a micronucleus or simply just an artifact. Targeted re-acquisition at high magnification can help to reduce the number of false positives.

Resources for QuickID Targeted Image Acquisition

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