Methods & Techniques
Methods
Please find below a selection of method papers that we recommend as a starting point for sample preparations:
ZEISS AXIOSCAN Z1 AUTOMATIC SLIDE SCANNER
Huyghe et al.: Multiplex Immunofluorescence Combined with Spatial Image Analysis for the Clinical and Biological Assessment of the Tumor Microenvironment, Jove protocols, 2023
This sample preparation of FFP sections is also suitable for any other confocal/ widefield microscope.
MILTENYI BLAZE ULTRAMICROSCOPE
Miltenyi clearing and staining protocols for different tissue types to image cleared samples in Ethyl cinnamate. The sample preparation for 3D imaging requires the optical clearing and immunostaining of opaque tissues with different properties/ molecular compositions which require bespoke protocols for the tissue clearing and staining to achieve optimal results.
Mouse Brain with retaining transgene derived fluorescent signals
Techniques
Please find below some useful technical instructions:
SETTING UP KOEHLER ILLUMINATION/ ADJUSTING THE CONDENSER
If you use the brightfield/DIC channel on any widefield or confocal or spinning disc microscope, you should set up Koehler Illumination to get the best contrast for your image. This is irrelevant for confocal/fluorescence microscopy. Please contact the facility team (microscopy@imm.ox.ac.uk) to get a practical demonstration on the instrument you wish to work on.
Tutorial and background information from Zeiss how to set up Koehler illumination