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Scanning Angle Interference Microscopy (SAIM)
Published in Qiu-Xing Jiang, New Techniques for Studying Biomembranes, 2020
Cristina Bertocchi, Timothy J. Rudge, Andrea Ravasio
Scanning angle interference microscopy is based on the interference of coherent light incident on the sample, with the same light reflected from the silicon surface (see Figure 4.1). This interference occurs due to the differing path lengths of light directly illuminating the sample, at some height above the silicon layer, and light reflected from the silicon layer, passing through the silicon oxide to finally hit the sample. This sets up a standing wave illumination pattern, which varies in intensity (or electric field strength) with height above the silicon surface and depends on the angle of incident light as will be outlined below. The surface-generated standing wave as a function of incidence angle and fluorophore z-position (nm) is indicated in Figure 4.2a.
Morin inhibits colorectal tumor growth through inhibition of NF-κB signaling pathway
Published in Immunopharmacology and Immunotoxicology, 2019
HT-29 cells were seeded in 96-well plates with 5000 cells/well and treated with 60 mg/L Morin, or TNF-α/Smac-mimetic/Z-VAD-FMK, which was used to induce cell necroptosis as positive control for 4 h. After incubation, cells were washed with phosphate-buffered saline (PBS), resuspended in 300 mL binding buffer solution and incubated for 10 min in the dark with 5 mL Annexin V-FITC and then cells were imaged under differential interference microscope (DIC) and fluorescence microscope.
Morphometric analysis of sperm used for IVP by three different separation methods with spatial light interference microscopy
Published in Systems Biology in Reproductive Medicine, 2020
Marcello Rubessa, Mikhail E. Kandel, Sierra Schreiber, Sasha Meyers, Douglas H. Beck, Gabriel Popescu, Matthew B. Wheeler
Ten microliters of sperm were added to a glass slide and the sample was drawn across the slide to spread it evenly. These slides were air-dried and stored at 4°C until analysis. Images were taken of the slides using the SLIM quantitative phase imaging (QPI) instrument described in (Liu et al. 2018). Quantitative phase imaging (QPI) (Popescu 2011) is a label-free, nondestructive imaging modality that has important biomedical applications (for a recent review, see (Park et al. 2018)). QPI techniques yield a phase rather than an intensity map, which allows for quantitative measurements on transparent specimens, such as unlabelled cells. In reproductive research, such systems have found fertile ground in applications characterizing whole embryos (Warger et al. 2007; Nguyen et al. 2017) as well as sperm (Balberg et al. 2017; Lee et al. 2018). For the present study we chose spatial light interference microscopy (SLIM), which is highly sensitive in both space and in time. Because SLIM uses white light, which averages speckles, and common path interferometric geometry, which insures phase stability, it is highly sensitive (Kandel et al. 2017). SLIM has been used recently to characterized the topography and refractometry of sperm (Liu et al. 2018; Rubessa et al. 2019). Images were manually segmented using the ROI feature in ImageJ (NIH, Bethesda, MD). To eliminate between-researcher variation, all images were manually annotated by the operator using the ROI feature in ImageJ (Figure 1). We found all parameters circumscribed by sharp refractive index contrasts, except the acrosome, which in 2D images appears as a dense bump at the tip of the sperm head. As outlined in (Kandel et al. 2018), dry-mass was calculated as a scaled sum of the halo-corrected phase values within a selection (ImageJ’s Integrated Density measure, NIH, Bethesda, MD). In-line with common selection criteria, we chose to analyze all imaged spermatozoa that displayed intact acrosomes.
Comparison of Surgically Excised Premacular Membranes in Eyes with Macular Pucker and Proliferative Vitreoretinopathy
Published in Current Eye Research, 2019
Stefanie R. Guenther, Ricarda G. Schumann, Felix Hagenau, Armin Wolf, Siegfried G. Priglinger, Denise Vogt
Phase contrast and interference microscopy was performed with a modified fluorescence microscope (Leica DM 2500, Wetzlar, Germany) at magnifications between 500 and 40,000. For photographic documentation, a digital camera was used (ProgRes CF; Jenoptik, Jena, Germany).