SUBJECT

Microscopic Techniques

lecture

master

Faculty of Science

Biology MSc

3

Semester 2

Spring semester

1. History of the microscope, from single lens to the compound microscope.

2. Optical basics, image formation, interaction of light with matter,lens abberations

3. Diffraction, interference, dispersion, polarisation.

4. The compound microscope, optical and mechanical parts, critical and Köhler illumination

5. Image formation, objectives and eyepieces. Abberation correction.

6. Magnification and resolution, wave optic, diffraction and resolution, Furier plane, conjugate planes.

7. Phase contrast microscopy.

8. Polarisation microscopy, birefringence, anizotropy, polarisers. Measurement of retardation.

9. Nomarsky differencial interference contrast microscopy, Wollaston prisms, Hoffmann’s modulation contrast microscope.

10. Fluorescence microscopy, excitation and light emission, epifluorescence microscope, dichroic mirror, light sources, fluorescence probes.

11. Confocal laser scanning microscopy, optical sectioning, 3D reconstruction.

12. Special microscopy tecniques, ratio imaging, TIRF, FRET, multifoton excitation, spectral imaging techniques.

13. Microphotography, conventional and digital imaging, CCD detectors.

14. Image processing, FFT, convolution, measurements.

15. Care and alignment of the microscope.

• John D. Bancroft, Marilyn Gamble: Theory and Practice of Histological Techniques, Elsevier Health Sciences, 2008