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 is our most capable system built on the Eclipse TiE Microscope (Nikon Instruments) providing stable time-lapse imaging with automatic focus correction system 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

C1 LSM (Nikon Instruments) Room 8.04j WTBB

The C1 is our workhorse confocal LSM based on the TE2000E inverted microscope (Nikon Instruments) providing stable time-lapse imaging with automatic focus correction system and an ORCA ER monochromatic camera for TIRF (total internal reflection fluorescence) microscopy. The Nikon C1 four laser scanning confocal with TIRF attachment, housed in a heated CO2 incubation chamber. This system supports multichannel fluorescence capture, time-lapse recording, photoactivation imaging and spectral unmixing (For details click: https://sites.google.com/view/flowimaginguol/imaging/imaging-equipment/laser-scanning-confocal).

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).

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

BioStation IM (Nikon Instruments) Room 8.04 WTBB

The BioStation IM Microscope (Nikon Instruments) widefield fluorescence live cell imaging microscope is a workhorse within the facility allowing for high-quality phase, and fluorescence assessments of cell lines over long time courses requiring a controlled atmosphere and high optical magnifications. Often this is used to more cheaply optimise research approaches and experimental time frames prior to moving these projects onto more expensive and complex technologies such as confocal microscopy looking at a more focused time course (For details click: https://sites.google.com/view/flowimaginguol/imaging/imaging-equipment/live-cell-imaging-systems).

IncuCyte Zoom (Sartorius) Room 9.04c WTBB

The IncuCyte Zoom (Sartorius) widefield fluorescence live cell imaging microscope is a workhorse within the facility allowing for high-quality phase, and fluorescence assessments of cell lines over time courses under a controlled atmosphere. 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/live-cell-imaging-systems).

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

Bioscreening Services

Offering a bespoke service tailored to your specific needs from a single plate to a whole library, whether you are looking to carry out your own screen or make use of our staff to carry out the work for you we have the expertise, equipment and experience to guide you through the whole screening process.

Please contact us to discuss your specific requirements.

We have a range of instruments available to facilitate the accurate, reproducible plate preparation for High Content work.

For plate set-up there is an automated cell counter the Countess and for plating out cells fluidX xrd380 dispensers
For the treatment of 96 well plates - adding ‘libraries’, etc. there is our liquid handling platform - Agilent Bravo
The ‘fluidX xrd380’ dispensers are also available to facilitate and speed up staining protocols

Facility Charges

Required Equipment and Facility Charges

Combined Facility Annual Registration and access charge £50/yr
Nikon A1 Confocal Microscope (LSM) - £29.84/hr (£24.10/hr 4 hr Discount Block Rate)
Nikon TiE Widefield Fluorescent Microscope - £15.25/hr
Nikon BioStation Fluorescent Live Cell Microscope (Dishes) - £2.79 /hr
Essen IncuCyte Live Cell Microscope (Plates) - £3.16/hr
Perkin Elmer Operetta High Through High Content Plate Microscope - £30.75/hr
BioTek Cytation 5 Multi-Mode Imaging Plate Reader - £2.83/run
Additional Staff Support for data Analysis or Acquisition on top of training or setting of instrument settings charge at £30/hr