1. *School of Computer and Information, Hefei University of Technology
Confocal microscopy with structured illumination and increased depth of focus 12th International Workshop on Research and Education in Mechatronics
Thema
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Dipl.-Ing. (FH) Philipp Heidingsfelder#1, Prof. Dr. Jun Gao*2, Prof. Dr.-Ing. Peter Ott#3
#Institute for Mechatronics, Optical Engineering Lab, Heilbronn University
*School of Computer and Information, Hefei University of Technology
© HS HN • Autor (2006) • xyz.ppt

2. Contents Motivation Confocal Microscopy Structured Illumination
Thema
Contents
Motivation
Confocal Microscopy
Structured Illumination
Optical System
Results
Summary and Outlook
© HS HN • Autor (2006) • xyz.ppt

3. Structured Illumination
Thema
Confocal Microscopy
Structured Illumination
Inspection of microcircuits and MEMS
Optical sectioning property
Reduction in the presence of flared and scattered light in the image 
High-end applications in biotechnologie
Improved axial and lateral resolution
Great complexity
Combine confocal microscopy and structured illumination with moderate effort
Motivation
Confocal Microscopy
Optical System
Results
Summary Outlook
Structured Illumination
© HS HN • Autor (2006) • xyz.ppt

4. Structured Illumination
Principle of Confocal Microscopy
Beam Splitter
F´T
Fobj
Sample
Defocused Plane
Sample
Detector
Exit Pupil z
Detector pinhole
Focal Plane
Tube Lens
Illumination Pinhole
Objective
Illuminating
Beam
Motivation
Confocal Microscopy
Optical System
Results
Summary Outlook
Structured Illumination

5. Structured Illumination
Principle of Structured Illumination
Projection of a sinusoidal fringe pattern on the sample  Conrast evaluation for defocussing the sample
[C. Kranitzky, C. Richter, C. Faber, M.C. Knauer, G. Häusler; Institute of Optics, Information and Photonics, University of Erlangen-Nuremberg]
Motivation
Confocal Microscopy
Optical System
Results
Summary Outlook
Structured Illumination

6. Structured Illumination
Signal Evaluation
For each z-position 4 shifts of the field stop
4 irradiance values for each pixel E1, E2, E3, E4
 Contrast Evaluation:
[C. Kranitzky, C. Richter, C. Faber, M.C. Knauer, G. Häusler; Institute of Optics, Information and Photonics, University of Erlangen-Nuremberg]
Motivation
Confocal Microscopy
Optical System
Results
Summary Outlook
Structured Illumination

7. Structured Illumination
CCD1 1 4
Slit
Period
Grating
Tube Lens
Tube Lens
Grating
Incoherent
Illumination
Beam Splitter
Exit Pupil
Objective
Piezo
Linear
Actor
Projected Grating in Focal Plane of CCD1 z
Sample
Motivation
Confocal Microscopy
Optical System
Results
Summary Outlook
Structured Illumination

8. Structured Illumination
Thema
For the simulation the Optical Transfer Function (OTF) of the Optical System is needed
Pattern Illumination
Entrance Pupil
Exit Pupil
Detector Plane
The Point Spread Function (PSF) is defined as O2 S2 ∆z
S2'
∆z'
O2' ∆W α α' O1
O1'
Beam Splitter S1
S1'
Object Plane
NA: Numerical Aperture m: Magnitude of the microscope ∆W: Optical Path Differnce (OPD) in the exit pupil circ: Circular-Function F : Fourier Transformation λ: Wavelength
Motivation
Confocal Microscopy
Optical System
Results
Summary Outlook
Structured Illumination
© HS HN • Autor (2006) • xyz.ppt

9. Structured Illumination
F {EP}
F {PSF}
IF -1{FS·OTF}I
Motivation
Confocal Microscopy
Optical System
Results
Summary Outlook
Structured Illumination

10. Structured Illumination
CCD1 1 4
Slit
Period
Grating
CCD2
Beam Splitter
Tube Lens
Grating
Tube Lens
Incoherent
Illumination
Beam Splitter
Exit Pupil
Objective
Piezo
Linear
Actor
Projected Grating in Focal Plane of CCD1 z
Sample
Focal Plane CCD2
Motivation
Confocal Microscopy
Optical System
Results
Summary Outlook
Structured Illumination

11. Structured Illumination
Simulated Focal Depth Response for focused (CCD1) and defocused camera (CCD2)
Motivation
Confocal Microscopy
Optical System
Results
Summary Outlook
Structured Illumination

12. Structured Illumination
CCD_1 1 4
Slit
Period
Grating
CCD_2
Beam Splitter
Tube Lens
Grating
Tube Lens
Incoherent
Illumination
Beam Splitter
Exit Pupil
Objective
Piezo
Linear
Actor
Focal Plane CCD_1
Projected Grating z
Sample
Focal Plane CCD_2
Motivation
Confocal Microscopy
Optical System
Results
Summary Outlook
Structured Illumination

13. Structured Illumination
Motivation
Confocal Microscopy
Optical System
Results
Summary Outlook
Structured Illumination

14. Structured Illumination
Light Source
CCD Camera
Grating
Step Motor and Piezo Actor
Tube Lens
Beam Splitter
Mirror
Objective
Motivation
Confocal Microscopy
Optical System
Results
Summary Outlook
Structured Illumination

15. Structured Illumination
Motivation
Confocal Microscopy
Optical System
Results
Summary Outlook
Structured Illumination

16. Structured Illumination
Step Height Standard: µm
Motivation
Confocal Microscopy
Optical System
Results
Summary Outlook
Structured Illumination

17. Structured Illumination
Softmark
Structured Illumination Mode
Confocal Microscopy Mode
Motivation
Confocal Microscopy
Optical System
Results
Summary Outlook
Structured Illumination

18. Structured Illumination
Summary
Interesting possibility to combine structured illumination with confocal microscopy.
Possibility to accelerate the measuring time with second camera.
Motivation
Confocal Microscopy
Optical System
Results
Summary Outlook
Structured Illumination

19. Structured Illumination
Outlook
Realization of the 2-camera-system
Downsizing the optical system
Analyzing the influence of the pattern illumination and compare it with the waveoptical model
Motivation
Confocal Microscopy
Optical System
Results
Summary Outlook
Structured Illumination

20. Structured Illumination
Thema
Thank you very much for your attention!
Motivation
Confocal Microscopy
Optical System
Results
Summary Outlook
Structured Illumination
© HS HN • Autor (2006) • xyz.ppt