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Light Imaging Section: Equipment
NAME |
BLDG |
RM |
TYPE |
MICROSCOPE |
Zeiss LSM 510 Meta* |
50 |
1435 |
confocal |
inverted |
Leica SP5 NLO* |
50 |
1536 |
confocal, |
inverted |
Leica DMR |
50 |
1536 |
wide-field |
upright |
Leica DMRI* |
50 |
1435 |
wide-field |
inverted |
Zeiss Axio Observer |
50 |
1423 (Skin Development Unit) |
wide-field |
inverted |
Leica TIRF |
50 |
1407 |
wide-field |
inverted |
Leica DMLB* |
10 |
13C121 |
wide-field |
upright |
PC Workstation* |
50 |
1533 |
|
|
Signup for the * instruments is done via a web-based calendar which can only be reached from inside the NIH network (http://niamsinside.niams.nih.gov/calendar/calcium3101/calcium310.pl?Op=UserLogin). For the other instruments, please contact us by e-mail or telephone. Also see the Access & Services section.
Zeiss LSM 510 Meta Confocal Microscope (bldg 50, rm 1435)
Zeiss LSM 510 Meta Confocal Microscope
The 510 Meta is based on an inverted Axiovert 200 microscope. In addition to "conventional" scanning it allows spectral imaging, a technique allowing the separation of fluorescent signals from fluorophores with emission spectra that overlap and are normally difficult to distinguish. It is equipped with 4 lasers (see Table) and several dichroic filters allowing numerous combinations of fluorophores. The 510 LSM microscope is surrounded by an environmental box to permit control of temperature, humidity, and CO2. It is also capable of autofocusing.
It is the instrument of choice for time-lapse work with live cells and techniques such as FRAP or FRET.
More information can be obtained on the Zeiss web site (http://www.zeiss.com/micro)
| Wavelength | Power (mW) | Type |
|---|---|---|
| 405 | 25 | diode |
| 458/ 477/488/ 514 | 40 | Ar |
| 543 | 1.5 | HeNe |
| 633 | 1.0 | HeNe |
Leica SP5 NLO Confocal Microscope (bldg 50, rm 1536)
Leica SP5 NLO Confocal Microscope
The SP5 is based on the Leica DMI6000 inverted microscope. It also allows spectral imaging. The basic equipment consists in 5 lasers (see Table). It has recently been upgraded to work in 2-or multi-photon mode with the addition of the MaiTai, a pulsed Ti:Sapphire laser. It is also equipped for intra-vital microscopy. Currently (June. 2008), it is used for fixed cell imaging in one-photon mode and for Second Harmonic Generation (SHG) and 2-Photon Excited Fluorescence (both in 2-photon mode).
More information can be obtained on the Leica web site (http://www.fluorescence-microscopy.com/)
A PC workstation (Dell) is loaded with the Zeiss image
viewing software and with the Metamorph software package (Universal
Imaging Corporation) for advanced digital image analysis. NIAMS users
also have access to the 3-D rendering and image analysis software Imaris
(Bitplane AG). Information on the specifics of the commercial software
packages IPLab,
Imaris, and Metamorph can be found on the websites of the parent
companies (Scanalytics, Bitplane, and Universal Imaging Corporation).
Non-commercial software packages which can be accessed freely by
everyone at NIH are mentioned with their URL in the Resources section
of this site.
| Wavelength | Power (mW) | Type |
|---|---|---|
| 405 | 50 | diode |
| 458/ 476/488/ 496/514 | 100 | Ar |
| 561 | 10 | diode |
| 594 | 2 | HeNe |
| 633 | 10 | HeNe |
| 690-1020 | 3,000 | Spectra-Physics Mai-Tai Ti:sapphire |
Leica DMR Fluorescence Microscope (bldg 50, rm 1536)
Leica DMR Fluorescence Microscope
The Leica DMR is an upright fluorescence microscope equipped with single bandpass filters for the observation of blue (DAPI, Hoechst), green (FITC, Alexa 488, GFP, YFP), red (Cy3, TRITC, Alexas 543,568, RFP), and deep red (Cy5, Texas Red, Alexa 592, 633, 647) fluo rescent conjugates. It also has double bandpass filters for the simultaneous observation of green and red fluorophores or of blue and yellow chromophores (CFP-YFP e.g.). It has 16x, 25x, 40x, 63x, and 100x oil immersion lenses and a 10x air lens. It can be used for fluorescence and transmitted light imaging in phase, DIC, bright-field, and pseudo-dark-field mode. It is fitted with a black and white digital camera (Hamamatsu ORCA) driven by the software IPLab (from iVision) on a Macintosh computer. A color digital camera (QImaging Micropublisher), driven by its own software, is also available.
The DMR is used principally for the examination of fixed cells /tissues on glass coverslips mounted on slides.
Leica DMRI Fluorescence Microscope (bldg 50, rm 1435)
Leica DMRI Fluorescence Microscope
The Leica DMRI is an inverted fluorescence microscope equipped with single bandpass filters for the observation of blue, green, red, and deep red fluorophores. It has 10x, 40x, and 63x air lenses. It is equipped with a LEP system of rotating filter wheels for 3-color time-lapse recordings and is also equipped with a Brinkmann micro-injection system. It can be used for fluorescence and transmitted light imaging in phase, DIC, and bright-field. Images can be recorded on a Coolsnap HQ (Photometrics) black and white digital camera or on the QImaging color camera.
The DMRI is used for checking expression of fluorescent proteins in cells grown in dishes with a coverglass bottom, for micro-injection into attached cells and for short timelapse recordings such as in chemotaxis experiments.
Leica TIRF System (bldg 50, rm 1407)
This system is based on the Leica DMI6000 microscope and is equipped with 4 diode lasers (405 nm, 488 nm, 561 nm, 633 nm) for up to 4 color-imaging. It is equipped with an Andor 512x512 EM CCD camera.
Microscope and EM CCD camera (lasers& monitors not shown)
100x, N.A. 1.46 TIRF objective
Leica DMLB Fluorescence Microscope (bldg 10, 13 C 121)
Leica DMLB Fluorescence Microscope
The Leica DMLB is an upright fluorescence microscope equipped with single bandpass filters for the observation of blue, green, red, and deep red fluorescent conjugates. It has 5x, 20x, and 40x dry lenses and can be used for fluorescence and transmitted light (phase and DIC). Images can be recorded on a Coolsnap HQ (Photometrics) black and white digital camera (using the IPlab software) or a QImaging color camera and software.
A 63x oil objective is available upon request.
The DMLB is recommended principally for the examination of fixed cells on glass coverslips mounted on slides.
PC Workstation (bldg 50, rm 1533)
A PC workstation (HP) is loaded with the full software versions of Zeiss LSM and Leica LASAF. NIAMS users also have access to the 3-D rendering and image analysis software Imaris (Bitplane AG) and the deconvolution software Huygens for confocal data (from SVI, supported by Bitplane). Information on the specifics of the commercial software packages IPLab, Imaris and Huygens deconvolution can be found on the websites of the parent companies (iVision, Bitplane, SVI). The non-commercial software packages Image J and MIPAV can be accessed freely by everyone at NIH. A scanner is attached to the workstation and is run via Adobe Photoshop under the import command.
Zeiss Axio Observer Fluorescence Microscope (bldg 50, rm 1423, Developmental Skin Biology Unit)
Zeiss Axio Observer Fluorescence Microscope
The Developmental Skin Biology Unit has acquired a Zeiss Axio Observer inverted microscope. It is equipped with long working distance lenses and uses a new contrast method (PlasDIC). PlasDIC gives excellent contrast in transmitted light with cells plated on plastic; the long working distance lenses can accomodate plastic dishes. This microscope therefore allows the examination and photography of fluorescently labeled cells on plastic dishes or plastic coverslips. One application is the visualization of fluorescent constructs for the selection of permanently transfected cell colonies. The system is equipped with filters for blue, green, and red fluorophores, can work in fluorescence or transmitted light modes (phase, plasDIC, bright-field), has 5x, 10x, 20x, and 40x air lenses and a Zeiss AxioCam digital camera. It is accessible to NIAMS staff.
Information on the PlasDIC technique can be found in the attached pdf's from the Zeiss website
Data Transfer
All computers, except that on the Leica SP5, are connected to the network. Users may use NIAMSIRPD/LIDATA/ Light Imaging Facility Users to transfer their files to their own computer. For the SP5, 1 GB flashdrives are available for file transfer.
Updated March 11, 2009
