University of Leeds Faculty of Medicine and Health Flow Cytometry and Imaging Facility

Bioimaging & Bioscreening Facility

Equipment

The facility has a range of light microscopes on the St James's University Hospital campus providing high resolution, fluorescence, high throughput-high content and live cell imaging monitoring capabilities. The facility currently houses 6 imaging systems and is constantly looking to resource new technologies that can help to improve our understanding of cell biology. All our equipment can be readily reserved thorough the QReserve online calendar system. Offline analysis of data can be carried out on any of our dedicated offline data analysis PC free of charge. All our microscopes are user-operated following specific training for the instrumentation from facility support staff. To organise training on a system please email: medlconf@leeds.ac.uk.

Laser Scanning Confocal Microscopes (LSM)

A1R LSM (Nikon Instruments) – Room 8.04j WTBB

The A1R system is built on the Eclipse TiE Microscope (Nikon Instruments) providing stable time-lapse imaging with automatic focus correction and a high speed motorized stage with a broad range of sample vessel options. The A1R LSM provides a hybrid scan head allowing for the collection of high-quality images through its unique pinhole and galvano scan head or high acquisition rates with 512 x 512-pixel definition at speeds of up to 30 frames per second. This system is fitted with its own incubation chamber allowing for live cell confocal assessments in gas and temperature controlled environment. The system has enhanced spectral imaging abilities and is able to perform fast acquisition of 32-channel spectral images and real-time spectral unmixing. Additional key features of this system include the incorporation of a perfect focus system and a fast piezoelectric focus drive. The microscope is equipped with 405nm, 457-514nm, 561nm and 642nm lasers. The system is controlled by the easy to use NIS-Elements C advanced software platform which enables complicated sequences of experiments to be performed such as photoactivation, image tiling (stitching of adjacent fields to give high resolution imaging of tissues) and multidimensional image acquisition (x, y, z, time, wavelength, and multipoint). For details click: https://sites.google.com/view/flowimaginguol/imaging/imaging-equipment/laser-scanning-confocal

LSM 980 with Airyscan (Carl Zeiss) Room 8.04j WTBB

LSM 980 with Airyscan 2 is the ideal platform for your confocal 4D imaging, optimised for simultaneous spectral detection of muliple weak labels with up to 900nm emisson with the highest light efficiency. The LSM 980 is built on a fully motorised Axio Observer 7 inverted microscope stand with Z-peizo, environmental chamber with temperature and CO2 control and uses the ZEN Blue 3 acquisition and analysis sortware. The Airyscan 2 includes options for fast imaging with improved resolution – resolution doubles up to 120nm from 240 nm resolution in confocal. In SR mode, the resolution can be further improved to 90 nm using the ZEN3 module Airyscan Joint Deconvolution. This woks in 2D and 3D image. Suitable applications include live cell imaging/time courses, colocalization studies, photo-activiation, FRAP, FRET, spectral imaging, stitching of large areas and imaging of fixed samples.

Available objectives: 2.5x, 10x, 20x, 40x and 63x

Available laser lines: 405nm, 445nm, 488nm, 514nm, 561nm, 594nm and 638nm

High Content High Throughput Fluorescence Microscopy

Operetta CLS (Perkin Elmer) Room 8.04d WTBB

With a choice of objectives and up to 4 channels per run the Operetta offers a way to collect images and data from 96 wells plates automatically with the option of a robotic plate handler for automatic running of large numbers of plates. Harmony and Columbus software offer meta-analysis capabilities (For details click: https://sites.google.com/view/flowimaginguol/imaging/imaging-equipment/throughput-highcontent-imaging).

CellDiscoverer 7 (Carl Zeiss) Room 8.04d WTBB

The CellDiscoverer 7 platform delivers fully automated image acquistion, analysis, and data management for robust phenotypic fingerprinting. The system has a range of excitations enabling the assessment of a variety of fluorescent tags. High through-put 96 well sampling capability with fully integrated robot plate loader and incubator allowing for a maximised automated assay capability. Automated high-content widefield inverted microscope with a full environmental chamber for temperature and CO2 control and Collibri LED illumination. The system is controlled by the Zen Blue software and is equipped with a large range of filter sets, seven LEDs (385 / 420 / 470 / 520 / 567 / 590 / 625 nm), four objectives (5x, 20x, 20x, 50x magnification chnager 0.5x,1x, 2x) and the Zeiss Axiocam 702 camera)

Widefield Fluorescence Microscopes

Eclipse TiE (Nikon instruments) Room 8.04f WTBB

The Eclipse TiE Microscope (Nikon Instruments) widefield fluorescence microscope is a workhorse within the facility allowing for high-quality phase, chromophore and fluorescence assessments of cell lines, and tissue in a wide variety of vessels. Often this is used to more cheaply optimise research approaches prior to moving these projects onto more expensive and complex technologies such as confocal microscopy (For details click: https://sites.google.com/view/flowimaginguol/imaging/imaging-equipment/widefield-fluorescence-micrscopes).

Live Cell Imaging Microscopes

HoloMonitor Live Cell Imaging System

The HoloMonitor system, is a live cell imager utilising holographic microscopy with the ability to measure cytometric parameters such as optical cell volume and thicknessso that cellular behaviour, drug response, and cell events can easily and continuously be visualised and quantified over time inside the incubator directly in the cell culture vessel. This enables HoloMonitor to automatically identify cells for quantitative single-cell and population analysis. The Holomonitor along with two proprietary software systems, Hstudio and HoloMonitor App Suite, automates the workflow from image capture, processing and, analysis to create publication-ready images data.

The HoloMonitor system comprises: an incubator-compatible base unit with motorised stage (optional), a laser unit, proprietary software.

Fluorescence, Absorbance & Luminescence Imaging

Cytation 5 (Biotek) room 5.04b WTBB

The system offers multi-mode detection for slide and plate formats with drug/reagent injection capabilities for fluorescence, absorbance and luminescence. Upgradable to offer microscopy with multi-mode fluorescence and phase outputs in z with environmental control. Gen5 Microplate Reader and Imaging Software allows for easy to use standard curve outputs from plate reader applications. Software includes functionality for easy image capture and analysis, for both qualitative and quantitative data including imaging deconvolution options. Capable of:

Qualitative and quantitative assays, endpoint, kinetic
EC50 determinations
Parallel line analysis
Spectral analysis
Micro-volume direct nucleic acid analysis

For details click:

https://sites.google.com/u/0/s/1b4NWrvniKo181IDjrwJ8ve2vhRfcOy-r/p/15wtjGl6XxKnnQ-CUKj3dBjryJM5Jyuib/preview

Services

We offer our researchers and our external collaborators a range of services and support to allow a streamlined workflow from assay design through to publication. The core services offered by the FoMH core Flow Cytometry & Imaging Facility are:

Project design
Grant Costings
Experimental Planning & Panel Design
Bioscreening
Applications Workshops
Equipment Training
Teaching Applications
Data Analysis Support
Experimental Troubleshooting
Outputs for Data Publication

Facility Charges

Required Equipment and Facility Charges

Zeiss LSM980 Confocal Microscope – £46.48/hr
Nikon A1 Confocal Microscope (LSM) - £46.48/hr (£40.00/hr 4 hr Discount Block Rate)
Nikon TiE Widefield Fluorescent Microscope - £18.08/hr
Perkin Elmer Operetta High Through High Content Plate Microscope - £24.75/hr
Zeiss Cell Discoverer 7 High Content imager – £46.48/hr
Phi Holomonitor Live Cell Imager – £2.89/hr
BioTek Cytation 5 Multi-Mode Imaging Plate Reader - £8.09/run
Additional Staff Support for data Analysis or Acquisition on top of training or setting of instrument settings charge at £30/hr