Combination of infrared laser heating through a microscope optics and heat shock response of cells achieves single-cell
gene induction in vivo. We will introduce a live-cell manipulation technique and show representative data in living
animals and plants. Further, we will discuss a local heating property in vivo through a temperature imaging technique
using a fluorescent thermometer.
Significance: A scene-based adaptive-optics (AO) system is developed and a method for investigating its imaging performance is proposed. The system enables derivation of Strehl ratios from observed images via collaboration with computer simulations. The resultant Strehl ratios are comparable with those of other current AO systems.
Aim: For versatile and noninvasive AO microscopy, a scene-based wavefront-sensing technique working on a Shack–Hartmann wavefront sensor is developed in a modal control system. The purpose of the research is to clarify the imaging performance of the AO system via the derivation of Strehl ratios from observed images toward applications in microscopy of living cells and tissues.
Approach: Two imaging metrics that can be directly measured from observed images (i.e., an energy concentration ratio and unbiased maximum ratio) are defined and related to the Strehl ratio via computer simulations. Experiments are conducted using artificial targets to measure the imaging metrics, which are then converted to Strehl ratios.
Results: The resultant Strehl ratios are >0.7 and 0.5 in the cases of defocus and higher aberrations, respectively. The half-widths at half-maximum of the AO-corrected bead images are favorably comparable to those of on-focus images under simple defocus aberration, and the AO system works both under bright-field illumination and on fluorescent bead images.
Conclusions: The proposed scene-based AO system is expected to work with a Strehl ratio of more than 0.5 when applied to high-resolution live imaging of cells and tissues under bright-field and fluorescence microscopies.
This letter proposes a method of configuring a testing target to evaluate the performance of adaptive optics microscopes. In this method, a testing slide with fluorescent beads is used to simultaneously determine the point spread function and the field of view. The point spread function is reproduced to simulate actual biological samples by etching a microstructure on the cover glass. The fabrication process is simplified to facilitate an onsite preparation. The artificial tissue consists of solid materials and silicone oil and is stable for use in repetitive experiments.
Adaptive optics is useful not only for the suppression of the blur of image, but also for the reduction of the aberration on the transmitted light. Recent years, methods for optical manipulation of biological tissue under the microscope is becoming available, whereas the live tissue often causes the considerable amount of optical aberration that prevents the clear convergence of the laser beam onto the target cell. This research shows the basic experiments to improve the convergence of the laser beam focused on the tissue in microscope by correcting the optical aberration using adaptive optics.
Live-cell imaging using fluorescent molecules is now essential for biological researches. However, images of living cells are accompanied with blur, which becomes stronger according to the depth inside the cells and tissues. This image blur is caused by the disturbance on light that goes through optically inhomogeneous living cells and tissues. Here, we show adaptive optics (AO) imaging of living plant cells. AO has been developed in astronomy to correct the disturbance on light caused by atmospheric turbulence. We developed AO microscope effective for the observation of living plant cells with strong disturbance by chloroplasts, and successfully obtained clear images inside plant cells.
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