Microscopy and Analysis Suite

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Advanced hypoxic imaging facility

Our hypoxic imaging facility consists of three state-of-the-art analytical instruments to study in vitro cell function under hypoxic conditions. Each instrument will measure different aspects of cell or molecular function under continuous hypoxic conditions. The facility includes a Leica Dmi8 Imaging Microscope, a BMG Labtech CLARIOstar plate reader with regulated oxygen control.

Benefits to cell function research

With this facility we can control oxygen levels to study different aspects of cell function from the level of a single cell through to entire cell populations. This will provide cell models that mimic oxygen levels in living systems. We use these cell models to explore the effects of oxygen deprivation, which can lead to tissue damage and disease.

This unique facility is funded by the University of Bath and a BBSRC ALERT14 equipment grant.

To use this facility please contact Ursula Potter.

Equipment

Baker Ruskinn Sci-tive Hypoxic workstation housing an advanced Leica Dmi8 imaging microscope

A Baker Ruskinn asymmetric Sci-tive Dual provides environmental control of O2, CO2, temperature and humidity in two inter-connected chambers. One chamber is available for cell culture under regulated oxygen conditions with HEPA filtration.

Features and applications (first chamber):

  • Large working area for mammalian cell culture
  • Large pass interlock that holds variety of flasks or dishes
  • O2 stability from anoxic to 20.9 % in 0.1 % increments
  • CO2 stability from 0.1 % to 30 % in 0.1 % increments
  • Humidity control from ambient to 85% RH
  • One touch sensor calibration
  • Easy access with Ezee sleeves
  • Cycle programming allows four different O2 and CO2 concentrations to set in a user defined timed sequence
  • Electrical power sockets in work area
  • HEPA filtration

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The second connected chamber houses a state-of-the-art motorized Leica Dmi8 imaging microscope with Andor iXon Ultra 897 EMCCD detection system and autofocus system. The two inter-connected chambers will enable the culture of cells and advanced imaging of cells under continuous hypoxic conditions.

Features and applications (second chamber):

  • Andor iXon Ultra 897 EMCCD detection system
  • Adaptive focus control (AFC) to maintain focus in real time
  • FRET measurements (YFP/CFP measurments)
  • FURA-2 calcium imaging
  • Temperature control
  • Automated focus control for long term measurements
  • Motorized microscope for remote control

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BMG Labtech CLARIOstar plate reader with regulated gas control

The BMG Labtech CLARIOstar plate reader is a multifunctional reader with regulated O2 and CO2 control that uses a revolutionary new dual monochromator technology that provides bandwidth selection and decreased background signal. To complement the high-resolution spatial imaging experiments provided by the Leica Dmi8 microscope, the plate reader provides exceptional performance in time resolved fluorescence, fluorescence resonance and FRET measurements for high throughput screening of biochemical-molecular and live cell assays.

Features and applications:

  • Uses advanced linear variable filter (LVF) monochromators and filters in single measurement
  • Adjustable bandwidth (8 – 100 nm) and wavelength (320 – 850 nm)
  • Ultrafast UV/Vis spectrometer
  • Automated Z focus control and adjustment of the height of the well
  • Integrated fluorophore library
  • Fluorescence polarization, FRET, BRET and time-resolved FRET measurements
  • Luminescence measurements
  • Atmospheric control unit for control of O2 (0.1-20%) and CO2 (0.1 – 20%) levels
  • Gas concentration curve display and LCD touchscreen control to regulate gas levels
  • Compatible with 6 to 1536-well plates
  • Top and bottom reading
  • End point and kinetic measurements
  • Sequential multi-excitation or multi-emission measurements
  • Spectral scanning (fluorescence, luminescence, absorbance)
  • Well scanning
  • Two built-in injectors with injection volume up to 500 μl
  • Temperature control

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