High Content Imaging and Analysis

Technology: LIPSI, NIS.ai, cell culturing services

High content imaging is about imaging more – more detection channels, more time points, more data. It’s no wonder that high content imaging and analysis is a powerful tool for so many applications, such as phenotypic screening in drug discovery research. However, the greatest strength of high content imaging – the volume of rich data created – can also be its greatest drawback, creating critical bottlenecks in experiment design, data storage, and image analysis. At the Nikon BioImaging Lab, our high content imaging services are centered around the LIPSI, a stable yet flexible platform for long-term live-cell high content that addresses common process bottlenecks to provide 24/7 operational efficiency.

High content imaging with LIPSI is great for applications ranging from cell health monitoring, to toxicology studies, and much more. For fast imaging thicker samples, LIPSI system, equipped with a set of long-range objectives, features Crest X-Light Series spinning disk confocal system for acquiring data deep in samples such as spheroids, organoids, organ-on-a-chip, and tissue. This system also features our NIS.ai deep learning software modules, which can be trained to perform a number of different analysis tasks, such as segmentation, and can be integrated into an automated imaging and analysis pipeline. Cell culturing services are also available.

Fluorescence Imaging

Technology: LIPSI, N-SIM S, AX / AX R, BioStudio-T, ECLIPSE Ti2-E

Fluorescence microscopy is widely considered to be one of the most sensitive imaging methods for detecting a given molecular target with high specificity. Fixed samples are generally labeled with dye-conjugated antibodies using standard immunohistochemistry/immunofluorescence techniques. Live samples can be made to genetically express a fluorescent protein fused to a target protein of interest or stained with membrane-permeable organic dyes.

Fluorescence underpins a number of important imaging methods providing a convenient mechanism for multichannel imaging via the labeling of different targets with spectrally-distinct fluorophores. All of our systems are built to perform fluorescence imaging, however there are many factors that must be considered to do so successfully. At the Nikon BioImaging Lab, we have an intimate understanding of concerns related to phototoxicity, sample preparation (including fluorophore selection), and other such critical factors that help determine the success of a fluorescence-based experiment.

Our imaging system are also capable of performing targeted FRAP and photostimulation experiments, FRET studies, and advanced 3D confocal and super-resolution imaging. We stand ready with the expertise to craft successful imaging, having a deep knowledge of fluorophores, microscope settings, mounting media, and related concerns.

Label-free Imaging

Technology: LIPSI, BioStudio-T, BioStudio-mini, Ti2-E Inverted Microscope, NIS.ai

While fluorescence imaging might be one of the most popular contrasting technique for optical microscopy, it is also one of the most invasive. Fluorophores are prone to photobleaching, limiting their useful lifetime, and resulting in the need for greater and greater illumination intensity to excite a similar amount of emission as the experiment progresses. This difficulty is compounded by the phototoxic response of live cells to high intensity illumination. Assays using sensitive cell types, such as stem cells, may not be completely compatible with fluorescence imaging of the desired target.

Cell health concerns are helping to fuel the renewed popularity of label-free imaging techniques, which help avoid both the need for high intensity illumination as well as perturbation of the system by the labeling process.

Artificial intelligence-based machine learning and deep learning analysis techniques are also helping to enable this trend. For example, the Convert.ai module (one of several NIS.ai deep learning software modules) automates the identification of key cell features from differential interference contrast (DIC) data, such as the simulated DAPI staining in the figure here. All of our systems support label-free imaging, with available techniques including new volume contrast, phase contrast, differential interference contrast (DIC), darkfield, and brightfield. Contact us today to see if label-free imaging will work for you.

High Throughput

Technology: LIPSI, AX R Confocal, ECLIPSE Ti2-E

Where high content imaging and analysis techniques are focused on acquiring and sifting through detailed multivariate data, high throughput is more focused on data volume, even if the dimensionality of that data is low. To be clear though, this doesn’t mean that high content imaging and analysis can’t be performed in a high throughput manner.

The LIPSI and ECLIPSE Ti2-E microscope feature a large 25 mm field of view (FOV) through the camera port, providing approximately twice the imaging area compared to competing microscope systems. Along with our large format cameras and the AX R confocal system, which are able to match the 25 mm FOV, these systems provide very high throughput. Throughput isn’t just determined by imaging area though, but also by acquisition rate. The Ti2-E inverted microscope features fast triggered acquisition, where a firing signal from the camera directly triggers the functions of system devices, bypassing slower software-based controls. The AX R confocal system has a fast resonant-scanning option, allowing for full-frame imaging at 15 frames per second (2048 x 1024). Along with our Denoise.AI deep learning-based denoising software module, you can achieve real-time imaging quality similar to a slow-scanning system.

Multi-point and large image stitching are straightforward to implement in your acquisition workflow in our NIS-Elements software with all the systems described here, making for easy whole-well, whole-slide, whole-vessel imaging.