Services

1. High-Content Imaging

High-Content Screening is automated image acquisition and analysis in high-throughput. It enables fast testing of many biological conditions including arrayed genetic and chemical screens, while ensuring reproducibility and objective quantification of images. A wide variety of different applications can be accommodated including cell counting and morphology, colony growth tracking, drug screening, cell viability and proliferation assays, protein translocation, and co-localization assays. Assays can be performed in multi-well format on fixed or live cells in monolayers or organoids. Time-lapse imaging for multiple plates using plate scheduler modules is available on the BioPipeline (44 plates) and the Incucytes (6 plates).

BioPipeline whole cerebral organoid embedded in a vascularized hydrogel, 4x, FITC (A. Yamand, Wrana lab, LTRI)

Incucyte confluency assay, phase with segmentation mask (K. Pacholczyk, Swallow lab, LTRI)

IN Cell Scratch Wound Healing Assay - 4x magnification, phase contrast (D. Ng, Swallow Lab, LTRI)

2. High-Content Data Analysis

We have several tools for automated image analysis. They include ones which can be used without extensive training to create simple analysis algorithms (Incucyte Analysis, Celígo Analysis and Columbus), as well as software.

NIS-Elements HC (Nikon)

NIS-Elements HC is a total acquisition-to-analysis solution for high-content imaging applications. It streamlines high-speed, automated multi-well plate acquisition, data review, analysis and management of multiple plate experiments. Multiple scans can be analyzed simultaneously during the imaging phase or run post-acquisition on offline stations. Heat maps of well/plates, sample images, binary masks, assay results, sample labels and other metadata are centralized for quick filtering, gating and drill down to cellular detail. NIS-Elements offers histogram, scatterplot, bar chart, XY line, classification and gating functions. Users can easily navigate within the Plate View and export to Excel or bitmap.

Celígo Analysis (Nexcelom Bioscience)

This built-in image analysis tool allows on-the-fly analysis of acquired images. Recommended applications include label-free cell growth tracking, cell cycle analysis, cell viability and apoptosis, transfection optimization, multiplex fluorescent expression assays, and many others. Once image acquisition is complete, data can be re-analyzed at an offline workstation. This instrument is ideal for up to 4-channel imaging of multi-well plates at 4X low-resolution.

Columbus (PerkinElmer)

Columbus is a universal image data storage and analysis system that enables access to stored images via a standard internet browser. The Columbus system allows scientists to remotely access, view, annotate, and analyze images. Columbus is based on an OMERO server and therefore supports a variety of existing microscope file formats. Integrated image analysis tools are intuitive and efficient. A high-performance computing server enables hyper-threading of image analysis. PhenoLOGIC™ machine learning technology provides an easy way to classify cells or subcellular regions within a given data-set.

Acapella (PerkinElmer)

Acapella is a powerful tool for image analysis. It has a set of proprietary scripts for detection of typical cell shapes and sub-cellular organelles. This is combined with a versatile programming environment to create customized algorithms for a variety of biological applications.

Matlab (Mathworks)

Matlab is the most powerful application for advanced analysis of images. Flexible image and data handling allows highly specialized routines for image analysis, including complex image filtering and segmentation. Costing for this analysis will be determined based on the complexity of the analysis.

Equipment

1. Nikon BioPipeline

The Nikon BioPipeline is a high-end, line scanning confocal high-content platform based on Nikon’s Eclipse fully-motorized, inverted Ti2 microscope and the A1R-HD25 system. An integrated 44-multi-well plate incubator provides multi-user assay scheduling. It is ideally suited for high-resolution fixed or live screening assays such as detection of DNA damage foci, translocation assays, measuring of cilia lengths, co-localization studies, drug screenings or any other assay that needs to combine high-resolution with automated imaging.

2. IN Cell Analyzer 6000

The IN Cell Analyzer 6000 is a laser-based, line-scanning confocal imaging platform with both widefield and confocal capabilities. This instrument is designed for high-throughput fixed or live imaging and assay development on a variety of vessel types. It can be used for timelapse and endpoint assays as well as 3-D imaging, co-localization studies and low signal assays.

3. Sartorius Incucytes

The Incucyte platforms are automated, high-content, widefield, multi-well plate acquisition, analysis and visualization platforms designed for long live imaging within an incubator. They offer dynamic insights into the health, viability, morphology, movement and function of cell models, with assays including confluence monitoring, spheroid and organoid detection, and scratch-wound. Our facility has an Incucyte S3 and the latest Incucyte SX5.

4. Celígo

Celígo is a low-resolution high-throughput microscope, equipped with LED-based light sources (brightfield and 3 fluorescent channels). Uniform well illumination and large-chip CCD camera allow users to image entire wells at a high-speed. Built-in image analysis tools allow performing on-the-fly analysis in a variety of biological applications (cell/colony counting, migrations assays, expression analysis and many others).

	Nikon BioPipeline Live	In Cell Analyzer 6000	Incucyte	Celigo
Type	Point-scanner confocal	Line-scanner confocal, widefield	Widefield	Widefield
Objectives	4X Nikon Plan Apo 0.2 NA 20 mm WD, Air 10X Nikon Plan 10X 0.45 NA 4 mm WD, Air 20X Nikon Plan Apo 0.75 NA 1 mm WD, Air 20X Nikon Apo Water 0.95 NA 0.95 mm WD, Water 20X Nikon S Plan Fluor ELWD 0.45 NA 8.2 mm WD, Air 40X Nikon Plan Apo 0.95 NA 0.17-0.25 mm, Air 40X Nikon Apo 1.15 NA 0.59-0.61 mm WD, Water 40X Nikon S Plan Fluor ELWD 0.4 NA 3.6 mm WD, Air 60X Nikon Plan Apo 1.2 NA 0.28-0.31 mm WD, Water 60X Nikon Plan Fluor Air ELWD 0.7 NA 1.8-2.6 mm WD, Air	4X Nikon Plan Apo 0.2 NA 20 mm WD, Air 10X Nikon Plan Apo 0.45 NA 4 mm WD, Air 20X Nikon Plan Fluor ELWD 0.45 NA 7.5 mm WD, Air 60X Nikon Plan Fluor ELWD 0.7 NA 1.8 mm WD, Air	4X Plan 10X Plan Fluor 20X Plan Fluor	4X
Excitation wavelengths	Blue 405 nm Green 488 nm Red 561 nm Far-red 640 nm	Blue 405 nm Green 488 nm Red 561 nm Far-red 640 nm	1st module: Green 440-480 nm (Incucyte S3, SX5) Red 565-605 nm (Incucyte S3, SX5) Swappable module Incucyte Green 453–485 nm Orange 648-674 nm NIR 576–639 nm	Blue 377+/-50 nm Green 483+/-32 nm Red 531+/-40 nm Far-red 628+/- 40 nm
Non-fluorescent channels	Brightfield	Brightfield, DIC, Phase	Phase	Brightfield
Imaging modality	2D, 3D	2D, 3D, Maximum intensity projection	2D	2D
Environmental Control	Yes	Yes	Yes	No
Associated software	NIS Elements	Columbus	Built-in Incucyte analysis	Built-in Celigo analysis

Pricing

Pricing is for external academic users only. Internal or industry users should contact Monica Hasegan (hasegan@lunenfeld.ca) for pricing.

HCS Instruments	Assisted	Unassisted (1st hour)	Unassisted (subsequent hours)	Offpeak¹ (1st hour)	Off-peak¹ (subsequent hours)
Nikon BioPipeline	$99.00	$75.00	$53.00	$25.00	$16.00
GE IN Cell 6000	$99.00	$75.00	$53.00	$25.00	$16.00
Celígo	$99.00	$30.00	$21.00	$10.00	$6.00

¹Peak hours are Monday to Friday, 9:00 AM to 6:00 PM

Incucytes	External Pricing ($/plate/day)
Essen Incucyte SX5	$110.00
Essen Incucyte SX3	$110.00

External Pricing ($/hr)
Software	Assisted	Unassisted
Image Analysis Workstation	$99.00	$12.00

Assisted Use	External Pricing ($/hr)
Training/Theory/Data Analysis/Assay Development	$99.00

Policies

Accessing the Facility

Users interested in accessing the High-Content Screening Facility should contact Monica Hasegan (hasegan@lunenfeld.ca) to discuss the scope of their project.

Getting Started

All new users must review our getting started guide. To work unassisted, users must first complete training with the facility manager.

Biosafety

Users who plan to do live cell imaging experiments have to meet with the manager to discuss potential bio-safety risks and to ensure appropriate training. Users must be familiar with practices outlined in Laboratory Biosafety Guidelines (3rd edition, 2004, Public Health Agency of Canada-PHAC and enforced by the human Pathogens and Toxins act).

Data Management

Data storage is the sole responsibility of the client and clients are urged to transfer data to their own storage devices as soon as possible. Data will be removed regularly from the instrument workstation since it adversely affects instrument performance.

Data storage on the dedicated analysis workstations will be removed when necessary (and without notice), so please use access to the LTRI network drives available on the analysis workstations to save your data or store directly to external storage devices.

Acknowledgement

Preferred acknowledgment for the Network Biology Collaborative Centre High-Content Screening Facility:
The authors wish to thank [staff name] of the Network Biology Collaborative Centre High-Content Screening Facility (RRID: SCR_025391) at the Lunenfeld-Tanenbaum Research Institute for [service]. The facility is supported by the Canada Foundation for Innovation and the Ontario Government.

Click here for our publication policy and preferred acknowledgement for other NBCC Facilities.

NBCC

Network Biology Collaborative Centre

Sinai Health, Lunenfeld-Tanenbaum Research Institute
600 University Avenue, Room 970
Toronto, Ontario Canada M5G 1X5

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High-Content Screening

An Introduction

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