Spatial light interference microscopy (SLIM)

Department of Electrical and Computer Engineering, Beckman Institute for Advanced Science & Technology, University of Illinois at Urbana-Champaign, Urbana, Illinois 61801, USA.
Optics Express (Impact Factor: 3.49). 01/2011; 19(2):1016-26. DOI: 10.1364/OE.19.001016
Source: PubMed


We present spatial light interference microscopy (SLIM) as a new optical microscopy technique, capable of measuring nanoscale structures and dynamics in live cells via interferometry. SLIM combines two classic ideas in light imaging: Zernike's phase contrast microscopy, which renders high contrast intensity images of transparent specimens, and Gabor's holography, where the phase information from the object is recorded. Thus, SLIM reveals the intrinsic contrast of cell structures and, in addition, renders quantitative optical path-length maps across the sample. The resulting topographic accuracy is comparable to that of atomic force microscopy, while the acquisition speed is 1,000 times higher. We illustrate the novel insight into cell dynamics via SLIM by experiments on primary cell cultures from the rat brain. SLIM is implemented as an add-on module to an existing phase contrast microscope, which may prove instrumental in impacting the light microscopy field at a large scale.

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    • "However, they present only off-axis reconstruction method in the results and the system's magnification is commanded principally for the physical dimensions. The flexibility of DHM offers the possibility of using it with another well established technique in order to develop a more robust measurement system [13] [14]. The Mirau Interferometer is normally used as an optical profiler when high magnifications are needed; this interferometer has some important features, such as spherical aberration compensation, easy difference optical path compensation, and insensitivity to external vibrations [15]. "
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    ABSTRACT: Digital Holographic Microscopy (DHM) typically uses either Michelson or Mach–Zehnder interferometers as interferometric tools to attain digital holograms. These interferometers need not only good optical alignment in order to compensate the spherical aberration, but also a special optical path difference compensation system when a low coherence illumination source is used. A Mirau interferometric objective appears as an alternative to overcome these difficulties and achieve reduced quadratic aberration in DHM. We show experimental results of the feasibility to perform DHM using this kind of objective; the tests conducted were in-line and off-axis configurations. In addition, we show not only the unique feature of the refocusing capability of DHM in a NBS 1963A resolution card showing its corresponding amplitude and phase images, but also a profile phase comparison of a 4.2 μm high micro lens using interferometry and DHM, extending the depth of focus of the microscope objective as proof of the proposal. As far as we know, this device has not been used in DHM.
    Optics and Lasers in Engineering 03/2013; 51(3):240–245. DOI:10.1016/j.optlaseng.2012.10.006 · 2.24 Impact Factor
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    • "The SLM can be used in both illuminating and imaging paths of the optical microscope, where it ensures a structured specimen illumination, or amplitude and phase modulation of the spatial spectrum, respectively [4]. Using these phase modulation techniques, the spatial light interference microscopy (SLIM) [5] [6] and the spiral phase contrast imaging [7] were presented as powerful techniques capable of measuring nanoscale structures and dynamics in live cells or enhancing standard phase contrast methods. A variability of operations provided by the SLM was also used to design the universal microscope, that operatively combines the standard techniques of optical microscopy [8] [9]. "
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    ABSTRACT: Incoherent correlation microscopy is recently discovered technique for digital imaging of three-dimensional objects in a quasi-monochromatic spatially incoherent light. Its operation is based on wavefront division carried out by a spatial light modulator and capturing correlation recordings of the observed scene. To achieve image reconstruction, at least a partial overlapping of the signal and reference waves created by the spatial light modulator is necessary. In the known experimental configurations, the overlapping of interfering beams is strongly reduced in off-axis areas of the object and the image can be reconstructed only in a very small portion of the field of view provided by the used microscope objective lens. Here, we propose and successfully demonstrate modified experimental system working with two-component relay optics inserted between the microscope objective and the spatial light modulator and providing full overlapping of correlated beams in all areas of the field of view of the objective lens. The benefits and applicability of the proposed system design are clearly demonstrated on the imaging of the USAF resolution targets.
    Journal of the European Optical Society Rapid Publications 02/2013; 8. DOI:10.2971/jeos.2013.13011] · 1.23 Impact Factor
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    • "Compared to other traditional optical techniques such as phase contrast microscopy or differential interference contrast microscopy, quantitative phase microscopy (QPM) has been developed to visualize and quantitatively analyze the distribution of phase shift of transmitted light through a specimen with nanometer resolution [1–3]. Since the amount of phase shift indicates the optical path difference (which contains the information of both the thickness and refractive index of the specimen), the QPM technique has been used to discern diverse cellular information under biophysical conditions such as the structural fluctuation of erythrocyte [4,5], cell growth depending on the cell cycle [6] and the measurement of refractive indices of intracellular materials [7,8]. In recent years, numerous novel techniques using QPM have been developed to enable a stable and quantitative measurement for long-term cellular dynamics using low-coherent illumination [7,8] and diffraction phase microscopy [9]. "
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