Instrumentation

The LSM-510-NLO is a confocal/two-photon system that allows for optical sectioning. It is used for 3D imaging of fluorescent proteins using one photon and/or two-photon excitation. Two-photon excitation allows for reduced phototoxicity and deeper penetration of light into the sample making it ideal for thick or live specimens. The microscope is equipped with the LSM-510 and NDD detection systems. The 510 portion consists of a 2 channel PMT setup for conventional fluorescence imaging and a META detector array for spectral separation of fluorochromes with overlapping emission curves. The NDD portion includes two non-descanned detectors allowing for maximum collection of emission and increased sensitivity. The microscope is equipped with an incubation chamber to control temperature as well as oxygen and carbon dioxide levels.

Laser lines: 458, 488, 514, 543, 561, 633, and a 720-1060 nm Chameleon laser for two-photon excitation

Microscope Location: 451G

Microscopy Center Contacts: Will Marshall Amanda Kroesen

Stowers Links:
Stowers Institute Home Page
Stowers Institute Core Facilities
Two-photon Microscopy
Place a Reservation
LSM-510-NLO Status

Web Links:
NLO Page: Zeiss
Multiphoton Microscopy: Mol Exp
Multiphoton Interactive Tutorial: Mol Exp

Literature:
Photoactivation with Two-Photon

Manuals:
Multiphoton Laser Scanning Microscopy
Chameleon
LSM-510 Meta
Axiovert 200
SMC Guidelines

The LSM-510-PRO is a confocal/two-photon system that allows for optical sectioning. It is used for 3D imaging of fluorescent proteins using one photon and/or two-photon excitation. Two-photon excitation allows for reduced phototoxicity and deeper penetration of light into the sample making it ideal for thick or live specimens. The microscope is equipped with the LSM-510 and NDD detection systems. The 510 portion consists of a 2 channel PMT setup for conventional fluorescence imaging and a META detector array for spectral separation of fluorochromes with overlapping emission curves. The NDD portion includes two non-descanned detectors allowing for maximum collection of emission and increased sensitivity.

Laser lines: 458, 488, 514, 543, 561, 633, and a 720-930 nm Chameleon laser for two-photon excitation

Microscope Location: 172

Microscopy Center Contacts: Sean McKinney Amanda Kroesen

Stowers Links:
Stowers Institute Home Page
Stowers Institute Core Facilities
Two-photon Microscopy
Place a Reservation
LSM-510-PRO Status

Web Links:
Two-Photon Microscopy: Zeiss
Multiphoton Microscopy: Mol Exp
Multiphoton Interactive Tutorial: Mol Exp

Literature:
LSM-510-PRO Brochure
Photoactivation with Two-Photon
LSM-510 Meta Manual
SMC Guidelines

The LSM-510-VIS is a confocal laser scanning microscope that allows for optical sectioning. The microscope is capable of imaging fluorescently labeled samples in 3D as well as taking transmitted light images that utilize various contrast methods for visualization. The detection system consists of two PMTs and one META detector array that can be used the separate fluorescent proteins with overlapping spectra.

Laser lines: 458, 488, 514, 561, 633, 405

Microscope Location: 451F

Microscopy Center Contacts: Richard Alexander Cindy Maddera

Stowers Links:
Stowers Institute Home Page
Stowers Institute Core Facilities
Confocal Microscopy
Place a Reservation
LSM-510-VIS Status

Web Links:
Confocal Microscopy: Zeiss
Confocal Microscopy: Mol Exp
Comparing Confocal and Widefield: Olympus

Literature:
LSM-510 Meta Manual
SMC Guidelines

The LSM-510-LIVE is a confocal laser scanning microscope that allows for optical sectioning and high speed imaging. The microscope is equipped with two detection systems. The 510 scan module consists of two PMTs for routine and 3D confocal imaging as well as a META detector array that can be used the separate fluorescent proteins with overlapping spectra. A PMT is also available for acquiring transmitted light images and utilizing various contrast methods for visualization. The LIVE detection unit uses a line-scanning approach as opposed to a rasterized point-scan to achieve ultrafast confocal imaging. It is capable of acquiring 120 512x512 images per second making FRAP experiments on fast moving proteins possible. The microscope is also equipped with an incubation chamber to control temperature as well as oxygen and carbon dioxide levels.

Laser lines: 458, 488, 514, 561, 633, 635, 405

Microscope Location: 172

Microscopy Center Contacts: Will Marshall Zulin Yu

Stowers Links:
Stowers Institute Home Page
Stowers Institute Core Facilities
Confocal Microscopy
Place a Reservation
LSM-Live Status

Web Links:
Confocal Microscopy: Zeiss
Confocal Microscopy: Mol Exp
Comparing Confocal and Widefield: Olympus

Literature:
LSM-510-DUO Manual
LSM-510 Meta Manual
SMC Guidelines

The Zeiss LSM 5 PASCAL is an upright confocal laser scanning microscope that allows for optical sectioning. The microscope is capable of imaging fluorescently labeled samples in 3D as well as taking transmitted light images that utilize various contrast methods for visualization. The detection system consists of two PMTs as well as a transmitted light detector.

Laser lines: 405, 458, 488, 514, 543

Microscope Location: 451F

Microscopy Center Contacts: Cindy Maddera Will Marshall

Stowers Links:
Stowers Institute Home Page
Stowers Institute Core Facilities
Confocal Microscopy
Place a Reservation
Upright Pascal Status

Web Links:
Confocal Microscopy: Zeiss
Confocal Microscopy: Mol Exp
Comparing Confocal and Widefield: Olympus

Manuals:
LSM 5 PASCAL
SMC Guidelines

The LSM-510-RGB is a confocal laser scanning microscope that allows for optical sectioning. The microscope is equipped with two detection systems. The 510 scan module consists of two PMTs for routine and 3D confocal imaging as well as a META detector array that can be used the separate fluorescent proteins with overlapping spectra. A PMT is also available for acquiring transmitted light images and utilizing various contrast methods for visualization. The microscope is also equipped with a ConfoCor 3 for taking fluorescence correlation spectroscopy (FCS) measurements. This technology is used to study diffusion times and protein-protein interactions. The ConfoCor 3 is equipped with high efficiency avalanche photodiodes (APDs) that allow for light imaging of dim samples.

Lasers lines: 405, 458, 488, 514, 561, 633

Microscope Location: 172

Microscopy Center Contacts:Shiqiang Dai Will Marshall

Stowers Links:
Stowers Institute Home Page
Stowers Institute Core Facilities
Confocal Microscopy
Fluorescence Correlation Spectroscopy
Place a Reservation
LSM-510-RGB Status

Web Links:
Confocal Microscopy: Mol Exp
Comparing Confocal and Widefield: Olympus
Fluorescence Correlation Spectroscopy: Zeiss

Literature:
LSM-510 Meta Manual
ConfoCor 3 Manual
SMC Guidelines

The LSM-710 is an upright microscope optimized for performing deep tissue, 2-photon imaging. Unlike the LSM-510, routine fluorescence acquisition is configured using a prism as opposed to an arrangement of emission filters. Signal from all emission wavelengths is then collected in parallel by a QUASAR detector. This detection system is similar to the 510 META but with significantly more sensitivity. Also attached to the LSM-710 are nine non-descanned detection (NDD) for collecting 2-photon emission and/or second harmonic generated signal, and a GaAsP detector for maximum sensitivity. Microscope function and image acquisition are controlled by Zeiss's new software package ZEN.  For long term live imaging an incubator has been added that offers temperature and CO2 control.

Lasers lines: 405, 458, 488, 514, 561, 633, and a 690-1064 nm Chameleon laser for two-photon excitation

Microscope Location: 176

Microscopy Center Contacts: Jeff Lange Will Marshall

Stowers Links:
Stowers Institute Home Page
Stowers Institute Core Facilities
Two-photon Microscopy
Place a Reservation
LSM-710 Status

Web Links:
NLO Page: Zeiss
Multiphoton Microscopy: Mol Exp
Multiphoton Interactive Tutorial: Mol Exp

Literature:
Photoactivation with Two-Photon
LSM-710 NLO Brochure
LSM-710 Training Guide
SMC Guidelines

The Leica SP2 is a confocal microscope that has a true UV laser. The optics are corrected for efficient UV excitation. Thus, this system is optimized for samples such as DAPI and HOESCHT. These dyes can also be excited using 405 nm lasers.

Microscope Location: 176

Microscopy Center Contacts: Shiqiang Dai Will Marshall

Stowers Links:
Stowers Institute Home Page
Stowers Institute Core Facilities
Confocal Microscopy
Place a Reservation
Leica SP2 Status

Web Links:
Leica Microsystems Homepage
Confocal Microscopy: Mol Exp
Comparing Confocal and Widefield: Olympus

Literature:
Leica SP2 Manual
SMC Guidelines

TThe PerkinElmer Ultraview is a spinning disk confocal microscope that allows acquisition of images at very high frame rates with minimum illumination of samples.  The microscope is equipped with one CCD (ORCA-R2) and one EMCCD (C9100-13) optimized for speed, sensitivity and resolution. Based on the Yokogawa CSU-X1 Spinning disk scanner, the system provides exceptional speed for live cell fluorescence imaging (up to 32 frames per second). A PhotoKinesis accessory is also available to target concentrated laser illumination onto a user defined region of interest to allow Fluorescence Recovery After Photobleaching (FRAP) and similar experiments.  Microscope function and image acquisition are controlled by PerkinElmer Velocity software. 

Microscope Location: 176

Microscopy Center Contacts: Zulin Yu Jeff Lange

Stowers Links:
Stowers Institute Home Page
Stowers Institute Core Facilities
Place a Reservation
Ultraview Spinning Disk Status

Web Links:
PerkinElmer
Spinning Disk Confocal Microscopy

Literature:
SMC Guidelines
PerkinElmer Ultraview Training Guide
PerkinElmer Ultraview Spinning Disk Manual
PerkinElmer Ultraview Spinning Disk Specifications

The Axiovert 451D is an inverted widefield microscope. It is capable of acquiring both fluorescence emission and transmitted light images that utilize various contrast enhancement methods such as DIC and Phase.  The system is equipped with a motorized stage and a high resolution Axiocam HRm (monochrome/black & white) camera as well as a Zeiss Apotome for acquiring fluorescent wide-field images in 3D.

Microscope Location: 451D

Microscopy Center Contacts: Qingfeng Yu Mark Richardson

Stowers Links:
Stowers Institute Home Page
Stowers Institute Core Facilities
Basics of Light Microscopy
Place a Reservation
Axiovert 451D Status

Web Links:
Basics in Optical Microscopy: Mol Exp
Apotome: Zeiss
ApoTome Tutorial: Zeiss
Axiocam HRm: Zeiss

Literature:
Axiovert 200 Manual
Structured Illumination
SMC Guidelines

The Axiovert 206 is an inverted widefield microscope. It is capable of acquiring both fluorescence emission and transmitted light images that utilize various contrast enhancement methods such as DIC and Phase. The system is equipped with a motorized stage and a high resolution Axiocam HRc (color) camera. The scope uses ONLY DRY objectives with long working distances that allow for imaging samples in plastic dishes/plates.

Microscope Location: 206

Microscopy Center Contacts: Cindy Maddera Shiqiang Dai

Stowers Links:
Stowers Institute Home Page
Stowers Institute Core Facilities
Basics of Light Microscopy
Place a Reservation
Axiovert 206 Status

Web Links:
Basics in Optical Microscopy: Mol Exp
Axiocam HRc: Zeiss

Literature:
Axiovert 200 Manual
SMC Guidelines

The Axioplan 176 Motorized is an upright widefield microscope. It is capable of acquiring both fluorescence emission and transmitted light images that utilize various contrast enhancement methods such as DIC and Phase. The system is equipped with a motorized stage and a high resolution Axiocam HRm (monochrome/black & white) camera.

Microscope Location: 176

Microscopy Center Contacts: Mark Richardson Qingfeng Yu

Stowers Links:
Stowers Institute Home Page
Stowers Institute Core Facilities
Basics of Light Microscopy
Place a Reservation
Axioplan 176 Status

Web Links:
Basics in Optical Microscopy: Mol Exp
Axiocam HRm: Zeiss

Literature:
Axioplan Manual
SMC Guidelines

The Axioplan 206 Motorized is an upright widefield microscope. It is capable of acquiring both fluorescence emission and transmitted light images that utilize various contrast enhancement methods such as DIC and Phase. The system is equipped with a motorized stage and a high resolution Axiocam HRc (color) camera. It is important to note that the microscope body has no motorized components and cannot be controlled by the software.

Microscope Location: 206

Microscopy Center Contacts: Cindy Maddera Shiqiang Dai

Stowers Links:
Stowers Institute Home Page
Stowers Institute Core Facilities
Basics of Light Microscopy
Place a Reservation
Axioplan 206 Std Status

Web Links:
Basics in Optical Microscopy: Mol Exp
Axiocam HRc: Zeiss

Literature:
Axioplan Manual
SMC Guidelines

The Low Light - TIRF system is used for widefield and TIRF imaging of dim samples. The system houses two specialized cameras: one for high sensitivity (EM-CCD) and another for high speed (Axiocam HS). Other additions to the system include a total internal reflection fluorescence (TIRF) illuminator used to generate ultra-thin optical sections of samples directly adjacent to the coverslip and an incubation chamber to control temperature as well as oxygen and carbon dioxide levels. The following laser lines are available for TIRF imaging: 458, 488, and 514.

Microscope Location: 451E

Microscopy Center Contacts: Richard Alexander Sean McKinney

Stowers Links:
Stowers Institute Home Page
Stowers Institute Core Facilities
Basics of Light Microscopy
Place a Reservation
Low Light Turf Status

Web Links:
Low Light TIRF: Zeiss
TIRF: Mol Exp

Literature:
Axiovert Manual
SMC Guidelines

The Tissue Culture Scope is used to quickly visualize samples in transmitted light with the option of using Phase to create contrast. The microscope has no fluorescence capabilities and is not equipped with a camera.

Microscope Location: 451F

Microscopy Center Contacts: Shiqiang Dai Cindy Maddera

Stowers Links:
Stowers Institute Home Page
Stowers Institute Core Facilities
Basics of Light Microscopy
Tissue Culture Status

Web Links:
Carl Zeiss: Homepage

Literature:
SMC Guidelines

The Leica Stereoscope 206 has a fluorescent excitation system with DAPI, CFP, GFP and RFP filter sets (a Cy5 filter is available in the cabinets in room 206 – please contact for assistance). The dissection scope is used to obtain low magnification images for large samples, such as whole mouse embyros. It is equipped with a 1.0X Plan-Apochromat Objective and a high resolution Axiocam HRc (color) camera.

Microscope Location: 206

Microscopy Center Contacts: Cindy Maddera Shiqiang Dai

Stowers Links:
Stowers Institute Home Page
Stowers Institute Core Facilities
Basics of Light Microscopy
Place a Reservation
Leica Stereoscope 206 Status

Web Links:
Basics in Optical Microscopy: Mol Exp

Literature:
Leica Stereo-Fluorescence Systems Manual
SMC Guidelines

The LSM-510-DEV is a development system that consists of a standard Zeiss 510 scan module and a variety of added features. The 510 detection unit contains two PMTs for routine and 3D confocal imaging as well as a META detector array that can be used the separate fluorescent proteins with overlapping spectra. A PMT is also available for acquiring transmitted light images and utilizing various contrast methods for visualization. Other detection systems on the microscope include a ConfoCorR 3 for taking FCS measurements, two NDDs for increased sensitivity, and a time correlated single photon counting (TCSPC) module for performing fluorescence lifetime imaging microscopy (FLIM). The microscope is also equipped with a two-photon laser that allows for reduced phototoxicity and deeper penetration of light into live or thick samples. Using two-photon the system is also capable of looking at second harmonic generation (SHG). The primary purposes of the DEV system is to work on the development of newer technologies by members of the Microscopy Center and perform more advanced experiments that cannot be carried out on other systems in the institute.

Lasers lines: 458, 488, 514, 543, 561, 633, and a 720-1040 nm Chameleon laser for two-photon excitation

Microscope Location: 461

Microscopy Center Contacts: Zulin Yu Richard Alexander

Stowers Links:
Stowers Institute Home Page
Stowers Institute Core Facilities
Two-photon Microscopy
Fluorescence Correlation Spectroscopy
Fluorescence Lifetime Imaging
Place a Reservation
LSM-510-DEV Status

Web Links:
NLO Page: Zeiss
Multiphoton Microscopy: Mol Exp
Multiphoton Interactive Tutorial: Mol Exp
Fluorescence Correlation Spectroscopy: Zeiss
Olympus: FLIM

Literature:
LSM-510 Meta Manual
ConfoCor 3 Manual
Chameleon Manual
Zeiss FLIM Manual
SMC Guidelines

The Zeiss PALM MicroBeam is a widefield system that provides a method for no-contact extraction and capture of cells for biochemical analysis. The system uses a fine-focused 355nm pulsed laser to cut-out and extract cell groups, single cells, or even parts of cells, without contamination or heat damage to the sample. PALM-LCM is equipped with a robotic arm that positions a micro-centrifuge tube cap above the stage to capture single or multiple samples once catapulted and can be used on a wide variety of sample types, including live, fixed, fluorescent, and brightfield.

Microscope Location: 172

Microscopy Center Contacts: Mark Richardson Zulin Yu

Stowers Links:
Stowers Institute Home Page
Stowers Institute Core Facilities
Place a Reservation
PALM Microbeam Status

Web Links:
Laser Microdissection: Zeiss

Literature:
PALM Microbeam Brochure
SMC Guidelines

The PALM (Photo Activation Light Microscopy) is a total internal reflection (TIR) based, widefield microscope. It's function is to acquire fluorescent images with resolution below the "diffraction-barrier" of common optical microscopes. PALM works by turning single photoactivatable fluorescent molecules on one at a time and finding their center. The molecules are then bleached to ensure that only one is activated per diffraction limited spot at any given moment.

Microscope Location: 451F

Microscopy Center Contacts: Sean McKinney

Stowers Links:
Stowers Institute Home Page
Stowers Institute Core Facilities
Place a Reservation
PALM Status

Web Links:
PALM: HHMI
TIRF: Mol Exp

Literature:
SMC Guidelines

The FluoroMax-3 spectrofluorometer is an instrument designed for fluorescence analysis in a cuvette. This system is equipped with a Xenon lamp and excitation monochromator for precise excitation of a sample and an emission monochromator and PMT (photo multiplier tube) detection scheme. The sample chamber can be equipped with a Peltier cooling device to ensure constant sample temperature during measurements. The instrument is controlled by the FluorEssence (Origin-based) software on the PC connected to the system. Examples of data that can be acquired on this system include excitation and emission curves of fluorescent species.

Microscope Location: 451C

Microscopy Center Contacts: Amanda Kroesen

Stowers Links:
Stowers Institute Home Page
Stowers Institute Core Facilities
Place a Reservation
Leica Stereo 451C Status

Web Links:
Product Page: Horiba Scientific
Principles of Fluorescence Spectroscopy: Horiba Scientific

Literature:
Zeiss FLIM Manual
SMC Guidelines