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Eunice Kennedy Shriver National Institute of Child Health and Human Development - Microscopy and Imaging Core
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Zeiss LSM 510 Inverted Meta Specifications

This point scanning instrument is available for either fixed slides or live specimen with temperature control, perfusion and solution changes. Photoactivation (405 nm), FRAP and FRET are fully supported. A Meta detector allows for spectral imaging and online emission fingerprinting.

Microscope

Zeiss Axiovert 200 M microscope (inverted)

Objectives

10x Zeiss plan-neofluar dry, 0.3 NA
20x Zeiss plan-apochromat dry, 0.75 NA
40x Zeiss plan-neofluar oil, 1.3 NA, DIC
63X Zeiss plan-apochromat oil, 1.4 NA, DIC
100x Zeiss alpha plan fluar oil, 1.45 NA, DIC

Fluorescence Filters Cubes (visual inspection only)

FITC / GFP: 450-490 nm excitation, 515-586 nm emission.
Rhodamin: 540-552 nm excitation, emission beyond 590 nm.
DAPI: 350-400 nm excitation, 417-477 nm emission.
Cy5: 610-650 nm excitation, 670-720 nm emission.

Laser Lines (scanning only)

405 nm: solid-state laser, 25 mW
458, 488 and 514 nm: Argon ion laser, 30 mW
543 nm: Helium-Neon laser, 1mW
633 nm: Helium-Neon laser, 15mW
AOTF switched: photoactivation, FRAP, FRET and uncagging fully supported

Filter Blocks (scanning only, must be used in certain combinations)

Excitation:

NT 80/20: beam splitter, 20% excitation reflected, 80% emission transmitted
HFT 405/488/543/633: dichroic 405, 488, 543 and 633 nm
HFT 405/488/543: dichroic 405, 488 and 543 nm
HFT 405/514: dichroic 405, 514 nm
HFT 458/514: dichroic 458, 514 nm
HFT 458/543: dichroic 458, 543 nm
HFT 458: dichroic 458 nm
HFT 488: dichroic 488 nm

Steering Meta vs PMT2/3:

NFT 490: long pass 490 nm
NFT 515: long pass 515 nm
NFT 545: long pass 545 nm
NFT 635 vis: long pass 635 nm
NFT KP 545: short pass 545 nm
Plate: glass plate correcting for misalignment, automatically inserted by software

Steering PMT 2 vs. PMT 3:

NFT 490: long pass 490 nm
NFT 515: long pass 515 nm
NFT 545: long pass 545 nm

Emission Filters

Meta detector

The Meta is a spectrometer spreading fluorescence emission among an array of detectors. It is, in itself, a configurable emission filter.

PMT 2

LP 420: long pass 420 nm
LP 475: long pass 475 nm
LP 505: long pass 505 nm
BP 420-480 IR: band pass 420 to 480 nm, IR-blocking
BP 470-500: band pass 470 to 500 nm
BP 475-525: band pass 475 to 525 nm
BP 505-530: band pass 505 to 530 nm

PMT 3

LP 505: long pass 505 nm
LP 560: long pass 560 nm
LP 650: long pass 650 nm
BP 500/20 IR: band pass 490 to 510 nm, IR-blocking
BP 505-530: band pass 505 to 530 nm
BP 505-550: band pass 505 to 550 nm
BP 530-600: band pass 530 to 600 nm
BP 560-615: band pass 560 to 615 nm

Detectors

Two photo-multiplier tubes fluorescence, 8 or 12 bit sampling (256 or 4096 gray levels, respectively)
One transmission detector for transmission / DIC imaging
One Meta detector for spectral imaging.. Up to 8 channels may be defined, each grouping any of the 32 detectors with a spectral width of 10 nm each (overlap between channels is not allowed). The meta detector is almost as sensitive as a PMT except at higher wavelength. However, its intrinsic noise is twice as high as a PMT.

Computer & Software

Fujitsu Siemens 2.4 GHz Xenon CPU running Windows 2000, 4 GB RAM, dual 114 GB storage, 2.4x DVD+/- recorder, dual 19" 1280 x 1024 flat panels
LSM 5 acquisition software with Physiology and Time Series modules