Indian Institute of Technology Delhi
Major Examination
PYL759: Computational Optical Imaging Attempt all questions
1. A fluorescence microscopy system is shown in the figure below. The illumination is with a blue
laser (405 nm) and the fluorescence excitation (530 nm) from the sample is being imaged.
(a) Interpret and explain the working of the system. [3 points]
(b) What is the significance of figure (B)? [3 points]
(c) The numerical aperture (NA) of the microscope objective lens (L6) is equal to 0.75. Estimate
the highest spatial resolution this system can achieve when a suitable image reconstruction
algorithm is used. [4 points]
2. (a) Show that the paraxial (Fresnel diffraction) field satisfies a modified wave equation [5
points]:
𝜕
(𝑖 + ∇2𝑥𝑦 + 𝑘 ) 𝑢(𝑥, 𝑦, 𝑧) = 0.
𝜕𝑧
(b) The transport of intensity equation is derived from the above equation and is given by:
𝜕𝐼
−𝑘 = ∇ ⋅ (𝐼 ∇ 𝜃 )
𝜕𝑧
Sketch a system that can perform measurements required for phase retrieval based on the
above equation. If the intensity of the wavefield 𝐼 is slowly varying across a transverse plane,
explain how you will computationally solve for the unknown phase 𝜃(𝑥, 𝑦) in that plane. [5
points]
3. An unknown complex-valued object 𝑔(𝑥, 𝑦) is illuminated by coherent X-rays followed by known
masks 𝑠𝑘 (𝑥, 𝑦) for 𝑘 = 1, 2, 3, … , 𝑁 and 𝑁 diffraction intensity patterns of the form
𝐼𝑘 = | 𝐹{ 𝑠𝑘 (𝑥, 𝑦)𝑔(𝑥, 𝑦)}|2 are recorded. The experimental configuration for one code-mask is
illustrated in the figure where 𝑔(𝑥, 𝑦) may be understood as the transmission function of the
sample.
Write a flow-chart of a phase retrieval algorithm that can use all the diffraction intensity patterns
𝐼𝑘 (𝑥, 𝑦) to estimate the amplitude and phase of the unknown test object. [10 points]
4. A system schematic shown below is claimed to perform single-shot 3D microscopy of fluorescent
particles over a depth range of 7x compared to the depth-of-focus of the system.
(a) Explain the working of the system as you understand it based on the schematic above. [3
points]
(b) What phase mask is likely being used in the pupil plane of the system? [3 points]
(c) Describe a possible algorithm that the system may be using for localizing particles in 3D
volume. [4 points]
5. (a) Define with equations what you mean by the MTF of an imaging lens. [ 5 points]
(b) Explain the figures below in terms of your understanding of the concept of MTF. [5 points]
6. (a) A typical camera involves a lens (like shown in problem 5) and an array sensor behind it. In a
second configuration, a random phase mask is placed in front of the lens aperture. Explain which
configuration is amenable to compressive sensing type image reconstruction (and thereby use of
much smaller number of detector pixels) and why. [5 points]
(b) Explain with appropriate reasoning why you cannot recover object phase from an in-line
hologram using the Fourier filtering algorithm. [5 points]
