The Center for Advanced Microscopy offers six high-resolution, point-scanning Confocal
Laser Scanning Microscope systems and one Yokogawa spinning disk Confocal Laser Scanning
Microscope system and provides a broad range of CLSM services for both biological
and physical sciences. Each system is designed for observation of a variety of live
or fixed biological specimens and a broad range of materials samples. Instrumentation
includes the systems listed below.
The 3i Spinning Disk confocal system is configured with a Yokogawa CSU-W1 T1 confocal
scanner that uses a microlens design to provide high-resolution confocal detection
with a wide imaging field of view. With imaging speeds of up to 200 frames per second,
the system has been optimized for rapid 2-D and 3-D imaging of live cells and tissues
over time with minimal photobleaching and phototoxicity. The system is configured
on an automated Zeiss Axio Observer inverted microscope with 3i SlideBook image acquisition
and analysis software and an OKO Lab environmental stage incubator for temperature
and CO2 control.
Multi-channel confocal fluorescence imaging, including standard green fluorescence
(excitation 488nm), red fluorescence (excitation 561nm), and far-red fluorescence
(excitation 637nm), as well as cyan fluorescence (excitation 445nm) and yellow fluorescence
(excitation 514nm).
2D and 3D live cell imaging and quantitative analyses over time
To get started using this microscope, contact Dr. Melinda Frame for help to collect data at operator assisted rates. Students can also become independent
users, but they must complete the NSC 837 Laboratory - Laser Scanning Confocal Microscopy
course, after which they are qualified to operate the microscopes at the self-operator
rate and are granted 24/7 access. Special arrangements can be made for faculty and
post-docs to attend classes.
Fees
Hourly rates for using this microscope are listed on our rates page.
The Leica Stellaris 5 Laser Scanning Confocal Microscope is high-resolution, high-sensitivity
confocal system configured on an automated inverted microscope with a supercontinuum
white light laser that provides a tunable range of laser excitation from 488nm to
685nm and both a 405nm and 448nm diode laser for imaging and photoactivation applications.
Fluorescence emissions are spectrally separated through a prism design and isolated
through variable slits that provide freely adjustable bandwidths in 1nm increments
through a detection range from 410-850nm. Four high-sensitivity Power HyD S detectors
are available for simultaneous or sequential recording of the confocal fluorescence.
The Stellaris 5 offers a range of advanced imaging applications including fluorescence
lifetime imaging (FLIM), imaging stitching for large area scanning, and Lightning
super resolution imaging with lateral (XY) resolution as low as 120nm and axial (Z)
resolution as low as 200nm.
Specifications
DMi8 Automated Inverted Microscope
Widefield fluorescence optics (blue, green, and red emission filters)
Fully automated precision scanning XY stage
Automated Z focus control, 0.05um step resolution
Brightfield, polarized light, and DIC transmitted light optics
Stage inserts that can accommodate most standard glass slides, 35mm dishes, chambered
slides, as well as multi-well dishes (6-, 24-, 48-, 96-well)
Leica Objectives
10x Plan Apo objective, NA 0.4, WD 2.56mm
20x Plan Apo objective, NA 0.75, WD 0.62mm
20x Plan Apo multi-immersion (oil, glycerol, water) objective, NA 0.75, WD 0.66mm
40x Plan Apo oil immersion objective, NA 1.3, WD 0.17mm
40x Plan Apo water immersion objective, NA 1.10, WD 0.62mm
63x Plan Apo oil immersion objective, NA 1.4, WD 0.14mm
100x Plan Apo oil immersion objective, NA 1.4, WD 0.13mm
Confocal Scanner
Two Galvano scan mirrors for high-resolution imaging, with uni-directional scanning
(1 frame/sec, 512x512) and bi-directional scanning (2 frames/sec, 512x512) options.
Fluorescence is detected through a single, variable pinhole aperture.
Maximum 8192x8192 pixel density
Detectors
Four high-sensitivity, low-noise Power HyD S photon counting detectors, providing
expanded detection sensitivity up to 850nm and the option of analog or photon counting
modes.
Each detector uses an adjustable variable slit for filter-free, spectral separation
of fluorescence through a 410nm – 850nm emission detection range. The variable slits
can be adjusted in as small as 1nm increments.
Dedicated transmitted light PMT detector for laser-based brightfield, polarized light,
or DIC imaging.
Lasers
Continuous wave 405nm diode laser, for most blue fluorochromes such as DAPI and for
targeted photoactivation and photobleaching applications.
Continuous wave 448nm diode laser, for most cyan fluorochromes such as CFP.
Pulsed supercontinuum white light laser, providing a tunable range of laser excitation
from 485nm-685nm in 1nm increments.
Software
Leica LAS-X acquisition and analysis software, version 4.6.0.
Applications
High-resolution, high-sensitivity confocal imaging in both fluorescence and reflected
light modes.
Simultaneous or sequential multi-channel confocal fluorescence imaging through a 410-850nm
emission detection range.
3-Dimensional image acquisition, rendering, and animation.
Time series acquisition and analysis.
Quantitative image analysis, including intensity, length, area, and co-localization
analysis.
Lightning Super Resolution mode for improved resolution that exceeds the diffraction-limit
of light, with lateral (XY) resolution as low as 120nm and axial (Z) resolution as
low as 200nm.
Fluorescence Lifetime imaging through the Leica TauSense Tools (TauContrast, TauSeparation,
TauGating, and TauInteraction). Provides fluorochrome lifetime information that can
be used to monitor changes in cellular microenvironments, improve spectral separation
of overlapping dyes, and remove unwanted autofluorescence.
Imaging stitching for large area scanning through the Leica Navigator mosaic tile
scan.
Targeted photobleaching and photoactivation applications, such as FRAP and FRET.
Excitation and emission lambda scanning and spectral linear unmixing for separation
of overlapping fluorescence emission spectra and separation and removal of background
autofluorescence. Emission lambda scanning employs a variable slit, with an emission
bandwidth down to 5nm. Excitation lambda scanning employs the variable white light
laser, with a laser range from 485-685nm and a bandwidth down to 1nm.
Brightfield or DIC transmitted laser-light imaging.
Training
To get started using this microscope, contact Dr. Melinda Frame for help to collect data at operator assisted rates. Students can also become independent
users, but they must complete the NSC 837 Laboratory - Laser Scanning Confocal Microscopy
course, after which they are qualified to operate the microscopes at the self-operator
rate and are granted 24/7 access. Special arrangements can be made for faculty and
post-docs to attend classes.
Fees
Hourly rates for using this microscope are listed on our rates page.
The Nikon A1Rsi Laser Scanning Confocal Microscope is a high-resolution, high-sensitivity
system configured on an automated inverted microscope platform with 6 lines of laser
excitation. A hybrid confocal scan head provides both a galvano scanner for high-resolution
imaging and a high-speed resonance scanner for rapid live cell imaging. Confocal
fluorescence is recorded using two standard photomultiplier tube (PMT) detectors and
two high-sensitivity Gallium Arsenide Phosphide (GaAsP) PMT detectors, with an independent
detector for the collection of brightfield, polarized light, and DIC images. In addition,
the system is configured with a high-sensitivity EMCCD camera for Total Internal Reflection
Fluorescence (TIRF) imaging and superresolution STORM applications, providing lateral
(XY) resolution as low as 10nm and axial (Z) resolution as low as 20nm. An OKO Lab
environmental stage incubator is available for temperature (heating/cooling) control
and CO2 control.
Specifications
Nikon Eclipse Ti Automated Inverted Microscope
Widefield fluorescence optics, including optics for blue, cyan, green, yellow, and
red fluorescence and multi-pass optics for blue, green, red, far red fluorescence
combinations and cyan, yellow, red fluorescence combinations.
X-Cite 120 LED fluorescent lamp
Fully automated precision XY stage
Automated control of microscope Z focus, 0.001um step resolution
MCL Nano-Drive piezo Z focus control, 0.001um step resolution, 100um focus range,
for rapid acquisition of 3D image series.
Perfect Focus System (PFS) to counter microscope focus fluctuations in real time during
long-term live cell imaging applications.
Brightfield, polarized light, and DIC transmitted light optics
Stage inserts that can accommodate most standard glass slides, 35mm dishes, chambered
slides, as well as multi-well dishes (6-, 24-, 48-, 96-well)
OKO Lab environmental stage incubator with heating and cooling temperature control
and CO2 control.
40x Plan Apo oil immersion objective, NA 1.30, WD 0.24mm
60x Apo oil immersion objective, NA 1.40, WD 0.14mm
60x Plan Apo water immersion objective, NA 1.20, WD 0.22mm
100x Plan Apo TIRF oil immersion objective, NA 1.45, WD 0.13mm
Confocal Scanner
Two galvano scan mirrors for high-resolution imaging, with uni-directional scanning
(2 frames/sec, 512x256) and bi-directional scanning (4 frames/sec, 512x256).
Two high-speed resonant scan mirrors for high-speed live cell imaging, with uni-directional
scanning (30 frames/sec, 512x256) and bi-directional scanning 60 frames/sec, 512x256).
Fluorescence is detected through a single, variable pinhole aperture.
Maximum 4096x4096 pixel density
Detectors
Two multialkali photomulitiplier (PMT) detectors (blue and far red fluorescence channels).
Two high-sensitivity Gallium Arsenide Phosphide (GaAsP) PMT detectors (cyan, green,
yellow, and red fluorescence channels).
A 32-channel spectral detector with spectral unmixing software for separation of overlapping
fluorescence emission spectra and/or background autofluorescence and for variable
band pass emission detection. The spectral detector can record a 430nm-750nm emission
range in 2.5nm, 6.0nm, or 10.0nm wavelength increments.
Dedicated transmitted ligth PMT detector for laser-based brightfield, polarized light,
or DIC imaging.
Andor iXon DU897 Ultra EMCCD camera for widefield, TIRFM, and STORM imaging.
Lasers
405nm diode laser, for most blue fluorochromes such as DAPI
445nm diode laser, for cyan fluorochromes such as CFP
488nm diode laser, for most green fluorochromes such as Alexa 488 or GFP
514 nm diode laser, for most yellow fluorochromes such as YFP
561nm diode laser, for most red fluorochromes such as Alexa 568 or RFP
647nm diode laser, for most far red fluorochromes such as Alexa 647 or chlorophyll
TIRF and STORM
The Nikon A1Rsi microscope system is also configured with optics and software for
Total Internal Reflection Fluorescence (TIRF) microscopy and 2D/3D super resolution
STORM imaging. For these specialized applications, the system is configured with
a high-sensitivity Andor iXon DU897 Ultra EMCCD camera (512×512 pixels, 16um-square
pixels, 17MHz) and a 100x Plan Apo TIRF oil immersion objective (NA 1.45). A 405/488/561/647
quadruple pass filter cube and a 445/514/561/ triple pass filter cube are available
to accommodate a wide range of fluorochromes for these applications.
Software
Nikon NIS-Elements AR acquisition and analysis software, version 5.42.06.
Applications
High-resolution, high-sensitivity confocal imaging in both fluorescence and reflected
light modes
Multi-channel confocal fluorescence imaging, including blue fluorescence (excitation
405 nm) through far red fluorescence (excitation 647 nm).
3-Dimensional image acquisition, rendering, and animation.
Live cell imaging and quantitative analyses over time
Quantitative image analysis, including intensity, length, area, cell counting and
co-localization analysis.
Imaging stitching for large area scanning
Targeted photobleaching and photoactivation applications, such as FRAP and FRET.
Spectral imaging and linear unmixing for separation of overlapping fluorescence emission
spectra and separation and removal of background autofluorescence using a 32-channel
spectral detector with 2.5nm, 6nm, and 10nm bandwidth selection.
Fluorescence protein imaging, including CFP, GFP, YFP, RFP, and BiFC analysis
Total Internal Reflection Fluorescence (TIRF) imaging.
Super resolution STORM imaging providing lateral (XY) resolution as low as 10nm and
axial (Z) resolution as low as 20nm.
Brightfield, polarized light, and DIC transmitted light imaging
OKO Lab environmental stage incubator for temperature (heating/cooling) control and
CO2 control.
Training
To get started using this microscope, contact Dr. Melinda Frame for help to collect data at operator assisted rates. Students can also become independent
users, but they must complete the NSC 837 Laboratory - Laser Scanning Confocal Microscopy
course, after which they are qualified to operate the microscopes at the self-operator
rate and are granted 24/7 access. Special arrangements can be made for faculty and
post-docs to attend classes.
Fees
Hourly rates for using this microscope are listed on our rates page.
The Nikon AXR CLSM system is configured on a fixed-stage upright microscope platform
with a focusing nosepiece and a high numerical aperture, long working distance immersible
(dipping) water objective. Designed with a large field of view (25mm), the system
is optimized for high-speed, high-resolution recording of fast cellular responses
in live tissues and organs.
Currently, the Nikon AXR confocal system is located in Dr. Brian Gulbransen's laboratory,
in BPS Room 3159. For access and training on the Nikon AXR, contact Dr. Melinda Frame within the Center for Advanced Microscopy.
Widefield fluorescence optics (blue, green, red, and far red emission filters)
Fully automated precision XY stage
Manual control of microscope Z focus
Ultra-fast automated piezo Z focus control, 0.001um step resolution, 450um focus range,
for rapid acquisition of 3D image series.
Brightfield transmitted light optics
Thermo stage plate for temperature control
Nikon Objective
25x Apo water immersible (dipping) objective, NA 1.1, WD 2mm
Single-position focusing nosepiece
Confocal Scanner
Two galvano scan mirrors for high-resolution imaging, with uni-directional scanning
(2 frames/sec, 512x256) and bi-directional scanning (4 frames/sec, 512x256).
Maximum pixel density for Galvano scan mode is 8192x8192.
Two high-speed resonant scan mirrors for high-speed live cell imaging, with uni-directional
scanning (30 frames/sec, 512x256) and bi-directional scanning 60 frames/sec, 512x256).
Maximum pixel density for Resonant scan mode is 2048x2048.
Maximum speed for Resonant scan mode is 720 frames/sec, 2048x16 pixels.
Fluorescence is detected through a single, variable pinhole aperture.
Detectors
Four detection channels: three high-sensitivity GaAsP PMTs plus one Multialkali PMT
designed for detection into the near-IR range.
Two tunable spectral detection channels (capable of spectral unmixing)
Two filter-based detection channels.
Dedicated transmitted light PMT detector for laser-based brightfield imaging.
Emission filters include 451/45nm band pass filter (Blue), 525/50nm band pass filter
(Green), 598/55nm band pass filter (Red), 700/75nm band pass filter (Far Red), and
795/110nm band pass filter (IR).
Lasers
405nm diode laser, for most blue fluorochromes such as DAPI
488nm diode laser, for most green fluorochromes such as Alexa 488 or GFP
561nm diode laser, for most red fluorochromes such as Alexa 568 or RFP
640nm diode laser, for most far red fluorochromes such as Alexa 647 or chlorophyll
730nm diode laser, for infrared (IR) fluorochromes such as Alexa Fluor 750
Software
Nikon NIS-Elements AR acquisition and analysis software, version 6.
Applications
Multi-channel confocal fluorescence imaging, including blue fluorescence (excitation
405 nm) through infra red fluorescence (excitation 730 nm).
3-Dimensional image acquisition, rendering, and animation.
Live cell imaging and quantitative analyses over time
Quantitative image analysis, including intensity, length, area, cell counting and
co-localization analysis.
Imaging stitching for large area scanning
Targeted photobleaching and photoactivation applications, such as FRAP and FRET.
Spectral imaging and linear unmixing for separation of overlapping fluorescence emission
spectra and separation and removal of background autofluorescence.
Fluorescence protein imaging, including CFP, GFP, YFP, RFP, and BiFC analysis
Brightfield transmitted laser-light imaging.
Training
To get started using this microscope, contact Dr. Melinda Frame for help to collect data at operator assisted rates. Students can also become independent
users, but they must complete the NSC 837 Laboratory - Laser Scanning Confocal Microscopy
course, after which they are qualified to operate the microscopes at the self-operator
rate and are granted 24/7 access. Special arrangements can be made for faculty and
post-docs to attend classes.
Fees
Hourly rates for using this microscope are listed on our rates page.
The Nikon C2+ Confocal Laser Scanning Microscope is a basic confocal system configured on an upright
microscope with four diode lasers and three high-sensitivity PMT detectors. This
system can be used for high-resolution fluorescence imaging of most fixed biological
specimens and materials samples such as metals and plastics. Reflection optics specifically
designed for material applications and EDF analysis are available. In addition, the
system is configured with a color camera for recording widefield fluorescence or conventional
transmitted light images, such as brightfield, phase contrast, polarized light, and
DIC.
Specifications
Nikon Eclipse Ni Upright Microscope
Widefield fluorescence optics (blue, green, and red emission filters)
SOLA SM II 365 fluorescence lamp
Manual XY Stage Control
Automated Z focus control, 0.001um step resolution
Brightfield, phase contrast, polarized light, and DIC transmitted light optics
DS-Fi2 color camera for recording conventional transmitted light and widefield fluorescence
images.
Nikon Objectives
10x Plan Fluor objective, NA 0.3, WD 16mm
20x Plan Apochromat Lambda objective, NA 0.75, WD 1.0mm
40x Plan Fluor Phase Contrast DLL objective, NA 0.75, WD 0.66mm
60x Plan Apochromat Lambda oil immersion objective, NA 1.4, WD 0.13mm
50x TU Plan Fluor objective, NA 0.80, WD 1.0mm, designed for reflected light applications
Confocal Scanner
Two galvano scan mirrors for high-resolution imaging, with uni-directional scanning
(1 frames/sec, 512x512) and bi-directional scanning (2 frames/sec, 512x512).
Fluorescence is detected through a single, variable pinhole aperture.
Maximum 2048x2048 pixel density
Detectors
Three high-sensitivity, low noise PMT detectors for confocal fluorescence.
Dedicated transmitted light PMT detector for laser-based brightfield, polarized light,
or DIC imaging.
Emission filters include 445/30BP (Blue), 525/50BP (Green), 600/50BP (Red), 660LLP
(Far Red), and 488/10 (Reflection).
DS-Fi2 color camera for widefield fluorescence or conventional transmitted light imaging.
Lasers
405nm Diode Laser, for most blue fluorochromes such as DAPI
488nm Diode Laser, for most green fluorochromes such as Alexa 488 or GFP
561nm Diode Laser, for most red fluorochromes such as Alexa 568 or RFP
640nm Diode Laser, for most far red fluorochromes such as Alexa 647 or chlorophyll
Software
Nikon NIS-Elements AR acquisition and analysis software, version 5.42.03.
Applications
High-resolution confocal imaging in both fluorescence and reflected light modes.
Simultaneous or sequential multi-channel confocal fluorescence imaging, blue through
far red fluorescence.
3-Dimensional image acquisition, rendering, and animation.
Time series acquisition and analysis.
Quantitative image analysis, including intensity, length, area, cell counting and
co-localization analysis.
Extended Depth of Focus (EDF) analysis, Topography analysis, Z height profiling.
Brightfield, polarized light, or DIC transmitted laser-light imaging.
Training
To get started using this microscope, contact Dr. Melinda Frame for help to collect data at operator assisted rates. Students can also become independent
users, but they must complete the NSC 837 Laboratory - Laser Scanning Confocal Microscopy
course, after which they are qualified to operate the microscopes at the self-operator
rate and are granted 24/7 access. Special arrangements can be made for faculty and
post-docs to attend classes.
Fees
Hourly rates for using this microscope are listed on our rates page.
The Olympus FluoView 1000 Spectral-based Laser Scanning Confocal Microscope is configured
on an automated inverted microscope platform with 6 lines of laser excitation. High-resolution
confocal fluorescence is detected through a single, variable pinhole aperture and
recorded using two high-sensitivity PMT detectors with with variable slits for spectral
wavelength selection and one high-sensitivity PMT detector with band pass emission
filters for wavelength selection. The unique Tornado scan permits rapid, high-intensity
photobleaching and photoactivation, optimized for methods such as FRAP, FRET, and
caged compound release.
Specifications
Olympus IX81 Automated Inverted Microscope
Widefield fluorescence optics (blue, green, and red emission filters)
X-Cite 120PC Mercury Arc Lamp
Fully automated precision XY scanning stage
Automated Z focus control, 0.01um step resolution
Brightfield, polarized light, and DIC transmitted light optics
Stage inserts that can accommodate most standard glass slides, 35mm dishes, and chambered
slides.
Olympus Objectives
4x U Plan FL N objective, NA 0.13, WD 17 mm
10x U Plan S Apo objective, NA 0.4, WD 3.1mm
20x U Plan FL N objective, NA 0.50, WD 2.1mm
40 U Plan FL N oil immersion objective, NA 1.3, WD 0.13mm
60x Plan Apo oil immersion objective, NA 1.4s, WD 0.15mm
100x U Plan S Apo oil immersion objective, NA 1.40, WD 0.13mm
Confocal Scanner
Two galvano scan mirrors for high-resolution imaging, with uni-directional scanning
(1 frames/sec, 512x512) and bi-directional scanning (2 frames/sec, 512x512).
Fluorescence is detected through a single, variable pinhole aperture.
Maximum 2048x2048 pixel density
Detectors
Two high-sensitivity PMT detectors with with variable slits for spectral wavelength
selection (capable of spectral unmixing).
One high-sensitivity PMT detector with band pass emission filters for wavelength selection.
Dedicated transmitted light PMT detector for laser-based brightfield, polarized light,
or DIC imaging.
Lasers
405nm diode laser, for most blue fluorochromes such as DAPI
458nm Argon gas laser, for cyan fluorochromes such as CFP
488nm Argon gas laser, for most green fluorochromes such as Alexa 488 or GFP
514 nm Argon gas laser, for most yellow fluorochromes such as YFP
559nm solid state laser, for most red fluorochromes such as Alexa 568 or RFP
635nm diode laser, for most far red fluorochromes such as Alexa 647 or chlorophyll
Software
Olympus FluoView FV10-ASW acquisition and analysis software, version 4.2.
Applications
High-resolution confocal imaging in both fluorescence and reflected light modes.
Simultaneous or sequential multi-channel confocal fluorescence imaging, blue through
far red fluorescence.
3-Dimensional image acquisition, rendering, and animation.
Live cell imaging and quantitative analyses over time.
Quantitative image analysis, including intensity, length, area, and co-localization
analysis.
Targeted photobleaching and photoactivation applications, such as FRAP and FRET.
Spectral imaging and linear unmixing for separation of overlapping fluorescence emission
spectra and separation and removal of background autofluorescence.
Fluorescence protein imaging, including CFP, GFP, YFP, RFP, and BiFC analysis
Brightfield, polarized light, or DIC transmitted laser-light imaging.
Training
To get started using this microscope, contact Dr. Melinda Frame for help to collect data at operator assisted rates. Students can also become independent
users, but they must complete the NSC 837 Laboratory - Laser Scanning Confocal Microscopy
course, after which they are qualified to operate the microscopes at the self-operator
rate and are granted 24/7 access. Special arrangements can be made for faculty and
post-docs to attend classes.
Fees
Hourly rates for using this microscope are listed on our rates page.
The Zeiss 980 CLSM with the AiryScan 2 super resolution module is a high-resolution,
high-sensitivity system designed for high-speed imaging with full spectral flexibility
and resolution. The system is configured on an automated inverted microscope platform
with 5 lines of laser excitation and the QUASAR spectral detection unit - a 32-channel
high-sensitivity Gallium Arsenide Phosphide (GaAsP) PMT detector that can be used
for multi-color fluorescence imaging through a 384nm-758nm emission range in partitions
as small as 20nm. In addition, the system is configured with the AiryScan 2 superresolution
detection module, designed to provide up to two times higher lateral (XY=120nm) and
axial (Z=350nm) optical resolution.
Specifications
Automated inverted Axio Observer 7 microscope
Widefield fluorescence optics (blue, green, and red emission filters)
X-Cite Xylis LED Fluorescence Lamp
Fully automated precision XY scanning stage
Automated piezo Z focus control, 0.01um step resolution, 500um focus range
Definite Focus 3 to counter microscope focus fluctuations in real time during long-term
live cell imaging applications.
Brightfield, polarized light, and DIC transmitted light optics
Stage inserts that can accommodate most standard glass slides, 35mm dishes, chambered
slides, as well as multi-well dishes (6-, 24-, 48-, 96-well)
Zeiss Objectives
5x Fluar objective, NA 0.25, WD 12.5mm
10x Plan-Apochromat objective, NA 0.45, WD 2mm
20x Plan-Apochromat objective, NA 0.80, WD 0.55mm
40x LD LCI Plan-Apochromat multi immersion (water, silicone oil or glycerin) objective,
NA 1.20, WD 0.41mm
63x Plan-Apochromat oil immersion objective, NA 1.40, WD 0.19mm
Confocal Scanner
Two galvano scan mirrors for high-resolution imaging, with uni-directional scanning
(2.5 frames/sec, 512x512) and bi-directional scanning (1.2 frames/sec, 512x512).
Fluorescence is detected through a single, variable pinhole aperture.
Maximum 8192x8192 pixel density
Detectors
Two multialkali photomulitiplier (PMT) detectors (typically for blue and/or far red
fluorescence emissions)
QUASAR spectral detection unit - a 32-channel high-sensitivity Gallium Arsenide Phosphide
(GaAsP) PMT detector that can be used for multi-color fluorescence imaging through
a 384nm-758nm emission range in partitions as small as 20nm.
The QUASAR spectral detector can also be used with spectral unmixing software for
the separation of overlapping fluorescence emission spectra and/or removal of background
autofluorescence. The spectral detector can record through the 384nm-758nm emission
range in 2.9nm, 4.4mm, 8.8nm, 17.6nm, 26.5nm, or 35.3nm wavelength increments.
AiryScan 2 superresolution detector, designed with 32 concentrically arranged GaAsP
detection elements, can provide up to two times higher lateral resolution (XY=120nm)
and axial resolution (Z=350nm). Addtional deconvolution software processing can produce
90nm lateral (XY) resolution and 270nm axial (Z) resolution.
Dedicated transmitted ligth PMT detector for laser-based brightfield, polarized light,
or DIC imaging.
Lasers
445nm diode laser, for cyan fluorochromes such as CFP
488nm diode laser, for most green fluorochromes such as Alexa 488 or GFP
514nm diode laser, for most yellow fluorochromes such as YFP
561nm solid state laser, for most red fluorochromes such as Alexa 568 or RFP
639nm diode laser, for most far red fluorochromes such as Alexa 647 or chlorophyll
Software
Zeiss ZEN acquisition and analysis software, Version 3.10.103
Multi-channel confocal fluorescence imaging, including cyan fluorescence (excitation
445 nm) through far red fluorescence (excitation 647 nm).
3-Dimensional image acquisition, rendering, and animation.
Live cell imaging and quantitative analyses over time
Quantitative image analysis, including intensity, length, area, and co-localization
analysis.
Imaging stitching for large area scanning
Targeted photobleaching and photoactivation applications, such as FRAP and FRET.
Spectral imaging and linear unmixing for separation of overlapping fluorescence emission
spectra and separation and removal of background autofluorescence using the 32-channel
QUASAR spectral detector
Fluorescence protein imaging, including CFP, GFP, YFP, RFP, and BiFC analysis
Super resolution AiryScan 2 imaging providing lateral (XY) resolution as low as 120nm
and axial (Z) resolution as low as 350nm.
Brightfield, polarized light, and DIC transmitted light imaging
Training
To get started using this microscope, contact Dr. Melinda Frame for help to collect data at operator assisted rates. Students can also become independent
users, but they must complete the NSC 837 Laboratory - Laser Scanning Confocal Microscopy
course, after which they are qualified to operate the microscopes at the self-operator
rate and are granted 24/7 access. Special arrangements can be made for faculty and
post-docs to attend classes.
Fees
Hourly rates for using this microscope are listed on our rates page.
A labeled plant cell co-expressing membrane markers of the endoplasmic reticulum,
golgi complex, and a soluble endoplasmic reticulum marker. Image taken on the Zeiss
Meta at the Center for Advanced Microscopy at Michigan State University. Research
from the lab of Dr. Federica Brandizzi.
The comprehensive range of advanced imaging applications offered by the Confocal Laser
Scanning Microscope systems include:
High-resolution, high-sensitivity confocal imaging in both fluorescence and reflected
light modes
Multi-channnel fluorescence imaging, including blue fluorescence (excitation 405 nm)
through far red fluorescence (excitation 730 nm)
High-speed imaging, through resonance scanning and spinning disk options.
3D rendering and animation
Co-localization analysis
Cell counting and image analysis software
Live cell imaging and quantitative analysis over time
FRAP, FRET, and BiFC imaging and analysis
Targeted photobleaching and photoactivation
Fluorescence protein imaging, including CFP, GFP, YFP, and RFP
Both excitation and emission spectral imaging
High resolution large area scanning
Fluorescence Lifetime Imaging microscopy (FLIM)
Total Internal Reflection Fluorescence (TIRF) microscopy
Super Resolution STORM imaging
Super Resolution AiryScan imaging
Super Resolution Lightning imaging
Extended Depth of Focus (EDF) analysis, Topography analysis, Z height profiling
For enrollment in NSC-837 Confocal Microscopy Lab, contact Dr. Melinda Frame, framem@msu.edu,
517-432-2327. This is a limited enrollment course with specific enrollment procedures.
Please see ourCoursespage for more information.
