# Optimizing for large-area imaging

**URL:** <https://forum.microlist.org/t/optimizing-for-large-area-imaging/3303>\
**Category:** Hardware\
**Tags:** confocal\
**Created:** [February 6, 2026, 5:34pm UTC](https://forum.microlist.org/t/optimizing-for-large-area-imaging/3303 "2026-02-06T17:34:49Z")\
**Posts on this page:** 2\
**Page:** 1

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**Author:** ![Amirali](https://avatars.discourse-cdn.com/v4/letter/a/e274bd/32.png) [@Amirali](https://forum.microlist.org/u/Amirali)\
**Post date:** [February 6, 2026, 5:34pm UTC](https://forum.microlist.org/t/optimizing-for-large-area-imaging/3303/1 "2026-02-06T17:34:49Z")

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Hi everyone,

I am working on a CyCIF project and need to image a large tissue area (approx. 2cm x 2cm). I am using an Olympus FV3000 point-scanning confocal.

In previous pilot runs on smaller areas, I used a 20x (NA 0.75) objective with the Galvano scanner. However, scaling this to 2cm x 2cm results in scan times exceeding 10 hours per round, which is not sustainable for a multi-round CyCIF workflow. The resolution I am imaging at is 1024x1024. I am doing Z-stacks with 5 steps in each image, but before processing we do a z-projection.

Is 10x magnification (NA 0.4) generally sufficient for MCMICRO/cell segmentation and immune phenotyping in tissue sections, or is the loss of NA too detrimental?

Also, do you have any suggestions on how to reduce the imaging time?

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**Author:** ![TEFK](https://avatars.discourse-cdn.com/v4/letter/t/3bc359/32.png) [@TEFK](https://forum.microlist.org/u/TEFK)\
**Post date:** [March 2, 2026, 9:47am UTC](https://forum.microlist.org/t/optimizing-for-large-area-imaging/3303/2 "2026-03-02T09:47:05Z")

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Dear Amirali,

An important consideration: what XY resolution do you really need? Do you scan at nyquist resolution? Perhaps you can zoom out, making larger pixels and speed up image acquisition? The Nikon Resolution Calculator I find a useful tool to assess XYZ resolution for point scanners.

Fast confocal imaging of large areas may be better done on a spinning disk confocal or slit scanner confocal (AION). Do you have access to any at your institute or nearby?

At low magnification the pinhole/ slit is often too large, reducing the ‘confocal effect’, but the imaging time is much shorter as trade-off. Especially on a dual camera system and if imaging deeper with water immersion objective your imaging time should drop to \< 1 hour.

Alternatively, point scanners with resonant capabilities may be useful too; especially if they do not require a minimal zoom so the confocal covers a large field of view at 2048x2048 pixel density.

Best,

Teo