Centre for Microscopy, Characterisation & Analysis
(formerly Biomedical Imaging and Analysis Facility)
The University of Western Australia
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Methods and techniques
Most of the methods and techniques detailed in the following pages are either processes that are currently in use in the BIAF or are the result of suggestions and discussions with other microscopists. Some information is derived from the confocal microscopy listserver and/or other sources and where possible, we have attempted to acknowledge the source. Attempts have been made to ensure the information is accurate. However, it is the responsibility of the individual user to determine if the methodolgy described here is suitable for their intended use.
Luminex Seminar Handout (2nd
May 2007) (pdf, 199KB) 
Luminex Run Setup Sheet
(pdf, 111KB)
Specimen Preparation
Autofluorescence removal (techniques to remove or minimise autofluorescence)
Nuclear DNA staining (Hoechst 33258 and 33342, DAPI)
Mounting medium recipe (PVA and glycerol antifade mounting medium)
Imaging non-adherent cells
Using the microscope
(These methods refer only to the Bio-Rad MRC1000/1024 UV Confocal Laser Scanning Microscope system located at BIAF.)
Collecting transmission images
Microscope calibrations (pixel sizing for scale bars)
Image Analysis and Digital Image Manipulation
Opening Biorad PIC files in Photoshop
Download Photoshop plugins to open Biorad PIC files
Creating Red/Green Merged images in Photoshop (This may aid in evaluating colocalisation.)
Colocalisation analysis (Lasersharp)
Creating montages (Confocal Assistant and Photoshop)
Other Sources of Information
Subscribing to the confocal microscopy listserver.
These pages are continually being revised. They may be reproduced provided the information remains complete and acknowledgement is given to the source. ©Dr Paul Rigby, CMCA.
This page was last revised on 15 May, 2007
