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Article ID: HXB2386 — Created: 25 Apr 2021 — Reviewed: 16 Jun 2021
Getting consistently great scans is sometimes a challenge depending on the specimen and slide preparation. High-magnification objectives tend to complicate the issue due to their narrower field of view and depth of field.
Following are several tips we have found valuable in helping to get the best scans from a uScope with a 40x or 60x objective. These tips are separated into various categories where software settings can help to improve your scans.
At 40x and 60x, both the Initial Only and Fast Stack focus methods are unlikely to produce satisfactory results. Predictive and Exhaustive focus methods are your best choices when it comes to well-focused scans.
Exhaustive Focus provides the best results since each field is individually focused. The down-side is the time required to scan. You can reduce the scan time by increasing the run focus step size and/or reducing the run range in the focus settings.
Predictive Focus provides very good results and is faster than the Exhaustive Focus method. The down-side is that a limited number of prediction fields are fully focused. A focus map is calculated and used for all other fields. You can alter the size of the grid used to capture prediction points. Smaller grids (4 or 5) typically yield better results since there are more fully focused prediction points.
The camera in the uScope provides a Standard Definition (SD) and a High Definition (HD) image mode. For best image resolution and quality, use the High Definition (HD) camera mode.
The uScope Navigator allows you to save images in either JPG or PNG image formats. Always use JPG unless you have a very good reason to use PNG.
It's a good idea to test your changes to the default settings. Each time you change settings, save the new (and old) settings into a profile.
Due to the potentially large size of images scanned with 40x or 60x objectives, we suggest you start with a small regions of interest. A ROI that is 10x10 (100) fields is a good starting point. Smaller ROIs let you see the results of your setting changes without taking a long time to scan.