Technologies
M. Others
Widefield Fluorescence Microscopy (WFF)
Conventional fluorescence imaging using broadband sources (LED, Metal-Halide, or Hg lamps) for full-field excitation.
Description
This training activity provides theoretical and hands-on instruction in the preparation and sectioning of biological samples using vibratomes and cryostats.
Participants learn how to select the most appropriate sectioning method according to the nature of the sample, the required section thickness and the subsequent microscopy or staining technique.
Vibratome Sectioning
Vibratome sectioning is used to obtain relatively thick sections from fresh, fixed or lightly embedded biological samples. Since the technique does not usually require paraffin or resin embedding, it can help preserve tissue morphology, enzymatic activity, fluorescence and antigenicity.
The training covers:
Sample fixation and preparation
Orientation and mounting of samples
Selection and installation of the appropriate blade
Adjustment of cutting speed, amplitude and section thickness
Sectioning of plant and animal tissues
Collection and handling of free-floating sections
Prevention of tearing, compression and vibration artefacts
Preparation of sections for histological staining, fluorescence microscopy and immunolocalization
Cryostat Sectioning
Cryostat sectioning is used to obtain thin sections from rapidly frozen biological samples. This method is particularly suitable when the preservation of lipids, enzymatic activity, fluorescence or antigenicity is required.
The training includes:
Sample collection and rapid freezing
Use of suitable embedding media for frozen samples
Orientation of samples in cryostat moulds
Temperature adjustment according to tissue characteristics
Preparation and trimming of frozen blocks
Sectioning and collection of cryosections on microscope slides
Prevention of folding, cracking and freezing artefacts
Storage and preparation of sections for staining, fluorescence microscopy and immunohistochemistry
At the end of the activity, participants will be able to select appropriate cutting conditions, obtain sections of suitable quality and identify the most common artefacts associated with vibratome and cryostat sectioning.