Abstract
Spatial light interference microscopy (SLIM) is a highly sensitive quantitative phase imaging method, which is capable of unprecedented structure studies in biology and beyond. In addition to the π/2 shift introduced in phase contrast between the scattered and unscattered light from the sample, 4 phase shifts are generated in SLIM, by increments of π/2 using a reflective liquid crystal phase modulator (LCPM). As 4 phase shifted images are required to produce a quantitative phase image, the switching speed of the LCPM and the acquisition rate of the camera limit the acquisition rate and, thus, SLIM's applicability to highly dynamic samples. In this paper we present a fast SLIM setup which can image at a maximum rate of 50 frames per second and provide in real-time quantitative phase images at 50/4 = 12.5 frames per second. We use a fast LCPM for phase shifting and a fast scientific-grade complementary metal oxide semiconductor (sCMOS) camera (Andor) for imaging. We present the dispersion relation, i.e. decay rate vs. spatial mode, associated with dynamic beating cardiomyocyte cells from the quantitative phase images obtained with the real-time SLIM system.
Highlights
Quantitative phase imaging (QPI) has emerged as a highly sensitive method for measuring nanometer scale pathlength changes induced by a specimen [1]
Further we have used a scientific-grade complementary metal oxide semiconductor camera (Andor) for imaging. This 5.5 Mega Pixel camera has smaller pixels that improve resolution by preserving Nyquist sampling and capable of imaging at 100 frames/s at full frame [36]. sCMOS has been expected to be used in a wide range of application fields which are currently dominated by Electron-Multiplication Charge Coupled Device (EMCCD) [37]
Real-time Spatial light interference microscopy (SLIM) of cardiomyocytes In order to show the capability of the real-time SLIM for quantitative imaging of dynamic cells, we have imaged beating cardiomyocyte cells
Summary
Quantitative phase imaging (QPI) has emerged as a highly sensitive method for measuring nanometer scale pathlength changes induced by a specimen [1]. This optical information yields biological information, including cell mass [2,3,4,5], membrane fluctuations [6,7], cell tomography [8,9,10,11], intracellular transport [12,13], tissue scattering [14,15,16], blood testing [17,18,19], cancer diagnosis [20]. Using broadband fields increases the spatial sensitivity of QPI (see, e.g., Chapter 8 in Ref. [1])
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