High Content Confocal Imaging

Opera
The Opera is designed for high-content, high-throughput imaging of multiwell plates. The multiple laser lines allow for multiplexed assays in up to four colors at magnifications of 10x-60x. The time of acquisition will vary based on number of channels, strength of signal and number of images per well. For example, a full 384-well plate using two channels and taking 2-3 images per well takes approximately 30 minutes to complete. The integrated automatic plate handler is capable of holding 42 plates (> 16,000 wells in 384-well format), with a capacity of approximately 30,000 wells per day.

The Acapella analysis software allows for automated image analysis on the fly and is capable of measuring multiple readouts including expression changes, translocation events, morphological changes and spot analysis. In addition, numerous features can be extracted from each analysis such as cell count, cell morphology, nuclear morphology, sub-cellular and texture features.


• Four laser excitation lines (405, 488, 561 and 647nm)
   Can be run in parallel
• UV lamp for non-confocal imaging in DAPI channel
• Capable of imaging plates in 96, 384 or 1536-well format.
• Magnifications range from 10x to 60x.
   10x, 20x and 40x air objectives
   20x, 40x and 60x water objectives for higher resolution
• Flexible image analysis software for creation of assay specific algorithms.



UltraView
The Ultraview is a high-content confocal imager designed for low-throughput experiments at higher resolution. Capable of acquiring images in up to 4 channels at magnifications between 20x-90x, the Ultraview provides the HTCB with the ability to handle samples in a variety of formats. The integrated Volocity software is designed for the acquisition and analysis of 3-D image stacks and time-lapse microscopy.  


• Four laser excitation lines (405, 488, 561 and 647nm)
   Can be run in parallel
• Real-time confocal imaging
• Capable of imaging using epifluorescence and transmitted light
• Can image slides, culture dishes, flasks and multiwell plates.
• 20x (air), 40x (oil) and 60x (oil) objectives
   Optovar module allows for additional 1.5x magnification
• Creation and viewing of 3-D image stacks and time-lapse movies
• 3-D and 4-D (time-lapse) analysis using Volocity

The Volocity software is designed for 3-D reconstruction of image stacks and creation of time-lapse movies.

Plate Reader Assays

Low content data acquisition is performed on Tecan InfiniTe plate readers. These can generate data in multiwell format for a number of different readouts. Fluorescent intensity can be measured at any wavelength for reporter assays. Luminescence can be measured in several different wavelengths for luciferase-based assays.



• Absorbance: UV Silicon photodiode, 4 channel parallel reading.
• Fluorescence: Extended wavelength (UV and red sensitive).
• Luminescence: Photon counting low dark current PMT 96-, 384-, 1536-well fibers.
• High energy UV/VIS/NIR xenon flash lamp, light–emitting diodes.

The plate readers are attached to our automated platforms for large-scale experiments or can be used as standalone units for single plate experiments.

Data Acquisition

For all readouts (low content, high content, 4-D), data can be provided in several ways. Investigators can choose to receive raw table files for each experiment. Alternatively, our informatics team can process the raw data into various formats. Please see the Informatics services for more details.