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Confocal Laser Illumination

High-intensity laser light source for confocal microscopy

Solid state lasers for high-performance laser excitation

With the increasing use of highly-complex cell-based 2D and 3D assays in biologic research, there is a pressing demand for improvements to the capabilities of automated high-content imaging. The ImageXpress® Confocal HT.ai High-Content Imaging System is a high-throughput solution that utilizes an 8-channel high-intensity laser light source to penetrate into deep tissue while reducing exposure times and increasing focus.

The illumination power of the laser light source significantly increases image intensity and assay sensitivity, which is especially important for dim samples. Imaging systems with lasers substantially decrease exposure times, which result in increased imaging speed and assay throughput. 3D imaging especially benefit from the laser light source with higher image intensities and improve image quality, which result in increased assay sensitivity and imaging speed.

 

**Data and images were acquired during development using customer samples. Results may vary. Highlighted features’ price, time to deliver, and specifications will vary based on mutually agreed technical requirements. Solution requirements may cause adjustment to standard performance.

 

AgileOptix technolgy with powerful, solid state light engine

AgileOptix technology makes it possible for the system to deliver the sensitivity and throughput needed for demanding applications by combining an advanced solid-state light engine with seven laser lines and eight filter combinations, proprietary spinning disk technology, state-of-the-art optics, and a scientific CMOS sensor.

Molecular Devices can successfully tailor the ImageXpress Confocal system to include customized software and hardware including the features described herein, as well as integration of other lab components such as incubators, liquid handlers, and robotics for a fully automated work cell. Check out our new Organoid Innovation Center where we showcase these cutting-edge technologies with novel 3D biology methods to address key challenges of scaling complex 3D biology.

Sales are subject to our Custom Product Purchase Terms available at www.moleculardevices.com/custom-products-purchase-terms.

Laser and LED light source specifications

The new modification of the ImageXpress Confocal HT.ai system equipped with multiline laser light source with matched filters spanning 405 nm to 730 nm excitation wavelength provides drastically increased illumination power to the sample. Laser light source results in brighter images, increased sensitivity, and increased speed and throughput speed of assays. The impact is especially important for the assays where sensitivity and timing are limiting factors.

Laser and LED light source specifications

 

 

Decreased exposure times with laser illumination

Confocal imaging and 3D image analysis are especially useful for capturing the complexity of 3D biological assays like organoids and spheroids. Here, we compared a standard imaging system with LED excitation to one with laser excitation.

 

 

Imaging 3D cancer spheroids

Imaging 3d cancer spheroids

A. DRAQ5 and Phalloidin-AF488 overlay of an untreated spheroid plus nuclear segmentation and area masks of the 2D projection image. Low magnification QC of spheroid staining could be used to locate treated and control spheroids in the well for accurate centering before acquiring at a higher magnification. B. Exposure times optimized to acquire images of equivalent brightness. Different exposures were required for different staining protocols. C. Speed of acquiring a 96-well spheroid plate in the ImageXpress Confocal HT.ai system at 20X using LED vs laser light sources.

overlay-of-an-untreated-spheroid

 

 

Imaging 3D lung organoids

Imaging 3D lung organoids

A. Confocal image of organoid culture (maximum projection), 10X. B. Images were taken using same exposure times for LED or laser light sources, then cell count and average intensities of objects (cells) shown for representative images. C. Exposure times were matched for 14-bit intensity range for both laser and LED. Optimized exposure times and duration of the experiment were substantially reduced by using lasers.

Confocal image of organoid culture

 

  • 3D Cell Models

    3D Cell Models

    3D cell cultures offer the advantage of closely recapitulating aspects of human tissues including the architecture, cell organization, cell-cell and cell-matrix interactions, and more physiologically-relevant diffusion characteristics. Utilization of 3D cellular assays adds value to research and screening campaigns, spanning the translational gap between 2D cell cultures and whole-animal models. By reproducing important parameters of the in vivo environment, 3D models can provide unique insight into the behavior of stem cells and developing tissues in vitro.

    Read More 

    Improve sensitivity, speed, and assay quality

    Complex Biological Assays

    Improve sensitivity, speed, and assay quality for complex biological assays

    Using a high-power laser light source, the system significantly increases light throughput to the sample, which results in brighter images, increased sensitivity, and increased assay throughput. The impact is especially important for the assays where sensitivity and imaging time are the limiting factors.

    Read application note 

  • Monitor lung organoids using 3D high content imaging

    Lung organoids as an assay model

    Organoid models have increasingly gained popularity in biologic research and screening to recapitulate complexity of real tissues. To model the in vivo human lungs, we have cultured primary human lung epithelial cells under conditions that promote the formation of 3D structures recapitulating the morphological and functional characteristics of the airway.

    Read application note 

    Organoids

    Organoids

    Organoids are three-dimensional (3D) multi-cellular microtissues that are designed to closely mimic the complex structure and functionality of human organs. Organoids typically consist of a co-culture of cells which demonstrate a high order of self-assembly to allow for an even better representation of complex in vivo cell responses and interactions, as compared to traditional 2D cell cultures.

    Learn More 

  • Organoid Innovation Center

    Organoid Innovation Center

    The new Organoid Innovation Center at Molecular Devices combines cutting-edge technologies with novel 3D biology methods to address key challenges of scaling complex 3D biology.

    The collaborative space brings customers and researchers into the lab to test automated workflows for organoid culturing and screening, with guidance from in-house scientists.

    Learn more 

    Using a laser light source for high-content automated imaging

    Using a laser light source

    In this study, we demonstrate improvement in assay sensitivity, precision, and speed of acquisition with a new configuration of the ImageXpress® Confocal HT.ai High-Content Imaging System that utilizes laser light source. The system drastically increases the power of illumination that results in brighter images, increased sensitivity, and increased speed and throughput speed of assays.

    Download scientific poster  

Resources for Confocal Laser Illumination

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