Mar 04, 2026
Brightfield image acquisition.
- Anastasia Filimontseva1,2,
- YuHong Fu1,2,
- Glenda Halliday1,2
- 1Brain and Mind Centre & Faculty of Medicine and Health School of Medical Sciences, The University of Sydney, Sydney, NSW 2050, Australia;
- 2Aligning Science Across Parkinson’s (ASAP) Collaborative Research Network, Chevy Chase, MD, 20815
Protocol Citation: Anastasia Filimontseva, YuHong Fu, Glenda Halliday 2026. Brightfield image acquisition.. protocols.io https://dx.doi.org/10.17504/protocols.io.eq2ly5bwpvx9/v1
License: This is an open access protocol distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited
Protocol status: Working
We use this protocol and it's working
Created: March 03, 2026
Last Modified: March 04, 2026
Protocol Integer ID: 245547
Keywords: ASAPCRN, slide scanner for brightfield microscopy, brightfield image acquisition, brightfield microscopy, automated slide scanner, image acquisition protocol, slideview vs200, using slideview vs200
Funders Acknowledgements:
Aligning Science Across Parkinson's
Grant ID: ASAP-020505
National Health and Medical Research Council Investigator Grant
Grant ID: 1176607
Abstract
Image acquisition protocol using SLIDEVIEW VS200 automated slide scanner for brightfield microscopy.
Troubleshooting
Creating the scan protocol
SLIDEVIEW VS200 automated slide scanner (Olympus Corporation, Tokyo, Japan) was used for automated image acquisition.
Sections were loaded into scan trays with the slide edges firmly pressing to individual slot sides.
These trays were loaded into the slide scanner.
A standardised imaging project was created for brightfield low-magnification overview and a higher magnification scan.
A separate scan project was created for sections undergoing different histological and immunohistochemical stains.
Adjusting scan parameters
The overview scan was performed using either a 2x objective (NA=0.06) or 10x objective (NA=0.4).
The exposure was manually adjusted to fit into the dynamic range of the camera.
The first slide was manually focused and this z-position was applied to all subsequent sections.
The higher magnification was performed using a 20x objective (NA=0.8).
The exposure settings were again adjusted to fit into the dynamic range of the camera, and the z-plane was set to 'Normal'.
The focus position density was also set to 'Normal'.
This was adjusted for each histological and immunohistochemical experimental batch of sections.
This scan project was saved and applied to all sections as a 'batch scan'.
Image acquisition
The low magnification overview scan was first run on all sections.
The overview scan generated a modifiable 'scan area' and 'focus map' that were adjusted if necessary before the higher magnification scan.