EVOS® FL Auto Imaging System

January 22, 2016

Oregon Health and Science University
Biomedical Engineering
Graduate Research Assistant

Overall

Performance

Ease-of-Optimization

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Company:

Life Technologies

Product Name:

EVOS® FL Auto Imaging System

Catalog Number:

AMAFD1000

The EVOS® FL Auto Imaging System is an easy to use bench top epifluorescence microscope that can be automated to scan multi-well plates, culture flasks and slides. The system features an enormous choice of LED/filter cubes that covers almost all of the current fluorescent dyes and fluorophores. We use the system extensively to scan tumor sections and 96-well plates for high content screening. Overall, the performance is excellent considering the relatively low price point when compared to other microscopes of similar capability.

Experimental Design and Results Summary

Application

High throughput content screening and imaging

Starting Material

Multi-well plates, IHC and IF slides

Protocol Overview

For cells plated on multi-well plates, we normally fix them 72 hours following treatment with 4% PFA or 100% methanol. After blocking in 2% BSA or goat serum, primary antibody staining is done overnight. We do secondary antibody staining for 1 hour the next day and scan right away. For tissue slides, we perform antigen retrieval in citric acid buffer in a pressure cooker for 10 minutes. Slides are permeabilized in 0.1% triton-x and blocked in 2% goat serum. Primary and secondary antibody stainings are the same as for multi-well plates.

Tips

Use the capture nominal focus feature before running a scan to save time on autofocusing and finding sample.

Results Summary

In the example below, the left image shows a full cross-section scan of a human breast tumor xenograft in nude mice. This slide has been staining with H&E by the histopathology core service at our university. The software comes with automated stitching feature. The right image shows another xenograft tumor section that has been stained for nuclei (blue, Hoechst 33342), HER2 (green, Alexa Fluor 488), and CD31 (red, Alexa Fluor 647). The average scan time for a section that spans 300 individual fields at 100X magnification is about 15 to 25 minutes, depending on the number of channels. Up to 4 LED/filter cubes can be installed in the system, which can be easily removed and swapped with a different one. In addition, there is no need to be in the dark since it comes with a stage cover that blocks ambient light.

Features Summary

Huge selection of LED/filter cubes that cover pretty much the entire spectrum of dyes from UV all the way to IR. Automated scanning features supporting most types of culture vessels, multi-well plates and slides.

Additional Notes

None

Image Gallery

Summary

The Good

Easy to use LED/filter cubes which require no warm-up time and have significantly longer life with consistent lightning.

The Bad

Image quality is not the best, especially at higher magnification. For high throughput scan, 100X to 200X work the best. There are better high magnification objective lens available but we have not tested them yet.

The Bottom Line

Overall, this system works surprisingly well. We have an Olympus IX-81 Scan^R system for side by side comparison and the performance is generally comparable at 100X and 200X magnifications. Considering the price point, which is about 1/4 of that of the IX-81, this microscope is of good value.

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