Tomographic phase microscopy with 180° rotation of live cells in suspension by holographic optical tweezers.
نویسندگان
چکیده
We present a new tomographic phase microscopy (TPM) approach that allows capturing the three-dimensional refractive index structure of single cells in suspension without labeling, using 180° rotation of the cells. This is obtained by integrating an external off-axis interferometer for wide-field wave front acquisition with holographic optical tweezers (HOTs) for trapping and micro-rotation of the suspended cells. In contrast to existing TPM approaches for cell imaging, our approach does not require anchoring the sample to a rotating stage, nor is it limited in angular range as is the illumination rotation approach. Thus, it allows noninvasive TPM of suspended live cells in a wide angular range. The proposed technique is experimentally demonstrated by capturing the three-dimensional refractive index map of yeast cells, while collecting interferometric projections at an angular range of 180° with 5° steps. The interferometric projections are processed by both the filtered back-projection method and the diffraction theory method. The experimental system is integrated with a spinning disk confocal fluorescent microscope for validation of the label-free TPM results.
منابع مشابه
Holographic Optical Tweezers Enable Super-resolution Localization Microscopy of Cells in Suspension
Super-resolution localization microscopy techniques such as dSTORM and PALM require several thousands of frames for the reconstruction of one single image and, thus, a long acquisition time [1]. This results in a high demand on the spatial stability of the sample, which is typically achieved by fixation and attachment to the coverslip. Treatment in such a way excludes e.g. super-resolved live-c...
متن کاملNanoscopy of bacterial cells immobilized by holographic optical tweezers
Imaging non-adherent cells by super-resolution far-field fluorescence microscopy is currently not possible because of their rapid movement while in suspension. Holographic optical tweezers (HOTs) enable the ability to freely control the number and position of optical traps, thus facilitating the unrestricted manipulation of cells in a volume around the focal plane. Here we show that immobilizin...
متن کاملOptical diffraction tomography for high resolution live cell imaging Citation
We report the experimental implementation of optical diffraction tomography for quantitative 3D mapping of refractive index in live biological cells. Using a heterodyne Mach-Zehnder interferometer, we record complex field images of light transmitted through a sample with varying directions of illumination. To quantitatively reconstruct the 3D map of complex refractive index in live cells, we ap...
متن کاملThe optical cell rotator.
The optical cell rotator (OCR) is a modified dual-beam laser trap for the holding and controlled rotation of suspended dielectric microparticles, such as cells. In contrast to optical tweezers, OCR uses two counter-propagating divergent laser beams, which are shaped and delivered by optical fibers. The rotation of a trapped specimen is carried out by the rotation of a dual-mode fiber, emitting ...
متن کاملmeasuring viscoelastic properties of Red Blood Cell using optical tweezers
Efforts have been made to study the behavior of complex materials in micrometer dimensions with various techniques. One of these methods is the use of optical tweezers for biophysical applications. Red blood cells, as the most abundant blood-forming cells, play an important role in the life of living organisms, and their unique mechanical properties are important. In this report, the study of s...
متن کاملذخیره در منابع من
با ذخیره ی این منبع در منابع من، دسترسی به آن را برای استفاده های بعدی آسان تر کنید
برای دانلود متن کامل این مقاله و بیش از 32 میلیون مقاله دیگر ابتدا ثبت نام کنید
ثبت ناماگر عضو سایت هستید لطفا وارد حساب کاربری خود شوید
ورودعنوان ژورنال:
- Optics letters
دوره 40 8 شماره
صفحات -
تاریخ انتشار 2015