Light sheet theta microscopy for quantitative imaging of large cleared samples

Author(s):  
Raju Tomer
2014 ◽  
Vol 127 (23) ◽  
pp. e1-e1
Author(s):  
R. S. Udan ◽  
V. G. Piazza ◽  
C.-w. Hsu ◽  
A.-K. Hadjantonakis ◽  
M. E. Dickinson

Author(s):  
E. Hämäläinen ◽  
J. Vattulainen ◽  
T. Alahautala ◽  
R. Hernberg ◽  
P. Vuoristo ◽  
...  

Abstract A novel technique and an instrument for on-line, quantitative imaging diagnostics and process control in thermal spraying have been developed and tested in laboratory and industrial conditions. In-flight spray particles are imaged by their natural luminosity with a short-exposure, digital CCD camera system. Particle images are processed using digital image processing techniques in a PC computer. The number of particles per frame and the spatially resolved particle velocities are calculated from the images. Spectrally resolved image information is further used to determine pyrometric two-color particle temperature. A number of different modes of data presentation have been developed. The developed instrument can be used to determine particle number, velocity and temperature distributions of individual in-flight particles from the imaged region of interest of the plume. Dividing the imaged area into smaller sections, spatial distributions of these parameters can be studied. SprayWatch system provides a technically simple, easy to operate, single imaging instrument, which can provide a visual overview of the spray plume in combination with quantitative evaluation of the most important spray particle parameters. In this paper examples of using the monitoring system with plasma and HVOF spraying are presented. Preliminary test results of using a semiconductor laser generated light sheet to detect cold particles is also demonstrated.


2018 ◽  
Vol 114 (3) ◽  
pp. 6a-7a
Author(s):  
Reto P. Fiolka ◽  
Kevin M. Dean ◽  
Meghan Driscoll ◽  
Erik Welf ◽  
Gaudenz Danuser

2015 ◽  
Vol 360 (1) ◽  
pp. 129-141 ◽  
Author(s):  
Francesco Pampaloni ◽  
Bo-Jui Chang ◽  
Ernst H. K. Stelzer

Development ◽  
2014 ◽  
Vol 141 (22) ◽  
pp. 4406-4414 ◽  
Author(s):  
R. S. Udan ◽  
V. G. Piazza ◽  
C.-w. Hsu ◽  
A.-K. Hadjantonakis ◽  
M. E. Dickinson

2015 ◽  
Vol 362 (1) ◽  
pp. 265-265 ◽  
Author(s):  
Francesco Pampaloni ◽  
Bo-Jui Chang ◽  
Ernst H. K. Stelzer

2021 ◽  
Author(s):  
Ilya Olevsko ◽  
Kaitlin Szederkenyi ◽  
Jennifer Corridon ◽  
Aaron Au ◽  
Brigitte Delhomme ◽  
...  

ABSTRACTFluorescence standards allow for quality control and for the comparison of data sets across instruments and laboratories in applications of quantitative fluorescence. For example, users of microscopy core facilities expect a homogenous and time-invariant illumination and a uniform detection sensitivity, which are prerequisites for quantitative imaging analysis, particle tracking or fluorometric pH or Ca2+-concentration measurements. Similarly, confirming the three-dimensional (3-D) resolution of optical sectioning micro-scopes prior to volumetric reconstructions calls for a regular calibration with a standardised point source. Typically, the test samples required for such calibration measurements are different ones, and they depend much on the very microscope technique used. Also, the ever-increasing choice among these techniques increases the demand for comparison and metrology across instruments. Here, we advocate and demonstrate the multiple uses of a surprisingly versatile and simple 3-D test sample that can complement existing and much more expensive calibration samples: simple commercial tissue paper labelled with a fluorescent highlighter pen. We provide relevant sample characteristics and show examples ranging from the sub-µm to cm scale, acquired on epifluorescence, confocal, image scanning, two-photon (2P) and light-sheet microscopes.Graphical abstractPyranine-labeled tissue paper, imaged upon 405-nm epifluorescence excitation through a 455LP LP dichroic and 465LP emission filter. Objective ×20/NA0.25. Overlaid are the normalised absorbance (dashed) and emission spectra (through line), respectively. In the present work we show that this “primitive” and inexpensive three-dimensional (3-D) test sample is a surprisingly versatile and powerful tool for quality assessment, comparison across microscopes as well as routine metrology for optical sectioning techniques, both for research labs and imaging core facilities.Research highlights-highlighter-pen marked tissue paper is a surprisingly powerful and versatile test sample for 3-D fluorescence microscopies-standard tissue paper presents features ranging from 400 nm to centimetres-our sample can simultaneously be used for testing intensity, field homogeneity, resolution, optical sectioning and image contrast-it is easy to prepare, versatile, photostable and inexpensive


2017 ◽  
Author(s):  
Bianca Migliori ◽  
Malika S. Datta ◽  
Christophe Dupre ◽  
Mehmet C. Apak ◽  
Shoh Asano ◽  
...  

AbstractAdvances in tissue clearing and molecular labelling methods are enabling unprecedented optical access to large intact biological systems. These advances fuel the need for high-speed microscopy approaches to image large samples quantitatively and at high resolution. While Light Sheet Microscopy (LSM), with its high planar imaging speed and low photo-bleaching, can be effective, scaling up to larger imaging volumes has been hindered by the use of orthogonal light-sheet illumination. To address this fundamental limitation, we have developed Light Sheet Theta Microscopy (LSTM), which uniformly illuminates samples from same side as the detection objective, thereby eliminating limits on lateral dimensions without sacrificing imaging resolution, depth and speed. We present detailed characterization of LSTM, and show that this approach achieves rapid high-resolution imaging of large intact samples with superior uniform high-resolution than LSM. LSTM is a significant step in high-resolution quantitative mapping of structure and function of large intact biological systems.


Author(s):  
Leslie M. Loew

A major application of potentiometric dyes has been the multisite optical recording of electrical activity in excitable systems. After being championed by L.B. Cohen and his colleagues for the past 20 years, the impact of this technology is rapidly being felt and is spreading to an increasing number of neuroscience laboratories. A second class of experiments involves using dyes to image membrane potential distributions in single cells by digital imaging microscopy - a major focus of this lab. These studies usually do not require the temporal resolution of multisite optical recording, being primarily focussed on slow cell biological processes, and therefore can achieve much higher spatial resolution. We have developed 2 methods for quantitative imaging of membrane potential. One method uses dual wavelength imaging of membrane-staining dyes and the other uses quantitative 3D imaging of a fluorescent lipophilic cation; the dyes used in each case were synthesized for this purpose in this laboratory.


Sign in / Sign up

Export Citation Format

Share Document