The Brown lab microscopes
Widefield fluorescence imaging workstation 1
Nikon TiE inverted motorized microscope with Perfect Focus System, phase contrast and epifluorescence; Prior Instruments linear motor motorized stage with linear encoders; TMC vibration isolation table; full line of high NA Plan Apo objectives; Andor Mosaic Digital Diaphragm multi-mirror array device with mercury arc lamp illumination for selective photobleaching and photoactivation of user-defined regions of interest; Andor MicroPoint laser ablation system with pulsed nitrogen pumped tunable dye laser; Andor Ultra 897 EMCCD and CoolSNAP HQ cooled CCD cameras on left and right side ports; Sutter Instruments excitation and emission filter wheels and shutters; Uniblitz shutter and driver for control of transmitted light; Photometrics DualView imaging module for simultaneous dual-wavelength imaging; X-Cite 120 LED illuminator; Okolab stage-top incubator with temperature and humidity control; Bioptechs objective heater and FCS2 heated imaging and perfusion chamber system; Warner Instruments in-line solution heater and heated imaging and perfusion chamber system with dual channel heater controller; PC workstation with 32GB RAM and MetaMorph software for device control and image acquisition, processing and analysis.
Widefield fluorescence imaging workstation 2
Nikon TE2000u inverted microscope with phase contrast and epifluorescence; mercury arc lamp illumination; TMC vibration isolation table; full line of high NA Plan Apo objectives; Princeton Instruments MicroMAX 512 BFT back-thinned frame transfer cooled CCD camera and Photometrics Quantix KAF1400 cooled CCD camera on left and right side ports; Uniblitz shutters and drivers for control of transmitted light and epifluorescence illumination; Bioptechs objective heater and FCS2 heated imaging and perfusion chamber system; Nevtek Air-stream incubator; Narashige hydraulic 3-axis micromanipulator and Medical Systems Corporation Pressure Injector; PC workstation with 4 GB RAM and MetaMorph software for device control and image acquisition, processing and analysis.
Nikon TE300 inverted microscope with phase contrast and epifluorescence; TMC vibration isolation table; Eppendorf NI2 InjectMan and FemtoJet Cell Injection system; Uniblitz shutters and drivers for control of transmitted light and epifluorescence illumination; Nevtek Air-stream incubator; Nikon cooled CCD camera; PC workstation with 4GB RAM and Nikon NIS Elements imaging software for shutter control and image acquisition, processing and analysis.
Spinning Disk Confocal imaging workstation in the Neuroscience Imaging Core
This microscope is a shared resource, located in the Neuroscience Imaging Core. Based around a fully motorized Nikon TiE inverted microscope equipped with DIC, LED transmitted and epi-fluorescent light sources, and Perfect Focus. An Okolab Bold Line stage-top incubator and controller with objective heater provide temperature, CO2 and humidity control. The objectives include: Plan Fluor 10x/0.3NA; Plan Apo 20X/0.75NA; Lambda S Apo LWD 40X/1.15NA; Plan Apo VC 60x/1.2NA; Plan Apo VC 100X/1.4NA. An ASI motorized piezo XYZ stage is equipped with rotary encoders for positional feedback and features a 22 nm XY step size with a 120 x 100 mm range of travel, and a 50 nm Z-axis step size with a 300 µm range of travel.
The Yokogawa CSU-W1 confocal unit is fully motorized and equipped with two camera ports, two 10-position emission filter wheels, a high efficiency motorized wide-field bypass mode, and a motorized adjustable aperture for detectors up to 17×16 mm. The spinning disk has 50 µm pinholes and operates at 4000 rpm allowing for imaging at up to 200 frames per second. A motorized slider contains three triple band pass dichroics for dual or triple wavelength imaging across the visible spectrum. The Revolution 500 series AOTF Laser Modulator and Multiport Combiner allows for fast switching between six solid state lasers: 100 mW 405nm diode laser; 50 mW 488nm DPSS laser; 40 mW 445nm diode laser; 50mW 515nm DPSS laser; 50 mW 561nm DPSS laser; 100 mW 640nm diode laser. A Precision Controller Unit allows for synchronized triggering of the cameras and laser blanking. On one camera port there is an Andor Neo 16-bit sCMOS camera with 2560×2160 pixel chip and 6.5 µm pixels deep-cooled to -40°C for fast confocal imaging with high spatial resolution and a large field of view. This camera has a 57% quantum efficiency and is capable of acquiring up to 30 frames per second. On the other camera port there is an Andor TuCam dual camera imaging adapter and two Andor iXon Ultra 897BV back-illuminated EMCCD cameras with 512×512 pixel chips and 16 µm pixels deep-cooled to -70°C for fast confocal imaging with maximum sensitivity. These cameras have a quantum efficiency of >90% and are capable of acquiring up to 56 frames per second. The TuCam contains two dual band-pass filter cubes with dichroic mirrors and emission filters for simultaneous dual-wavelength imaging of CFP/YFP or GFP/mCherry.
A laser-galvo operated FRAPPA unit introduced through a back port of the microscope allows for photobleaching or photoactivation of user-defined regions of interest within the field of view, with fast switching between any of the six available laser lines. The entire system is controlled by a PC workstation with 32GB RAM, 27″ LCD monitor with 2560×1440 resolution, nVidia high performance graphics processor, and MetaMorph Premier software for multi-dimensional acquisition, processing and analysis.