Zeiss LSM510 META (Inverted)

A confocal laser point scanning microscope is equipped with a high-precision motorized stage with “MultiTime” macro programing. The confocal detectors include two single channel PMTs and a META, polychromatic 32-channel detector for spectral imaging/emission fingerprinting. The META detector can also be used as another detector with freely definable emission ranges. Thelaser module includes7 lines covering wide ranges of excitation wavelength, 405nm, Argon laser (458, 477, 488 and 514nm lines), 561nm DPSS and HeNe 633 nm laser. The objective lenses include a Plan Apochromat 10x/0.45 NA, Plan Apochromat 20x/0.8 NA, C. Apochromat 40x/1.2 NA water and C. Apochromat 63x/1.2 NA water (Nomarski DIC, and Alpha Plan-Apochromat 100x/1.46 NA oil). The PC was updated in 2015 to Windows 7 and runs on Zen 2009 software.

Objectives

  • EC Plan-Neofluar 10x/0.3 NA
  • Plan-Apochromat 20x/0.8 NA
  • C. Apochromat 40x/1.2 NA water
  • C. Apochromat 63x/1.2 NA water
  • Plan-Apochromat 100x/1.46 NA oil

 

Applications

Optical sectioning and 3-D Reconstruction

2-D projection of Z-stack images: C. elegans dauers expressing lag-2::GFP in the inner-labial neurons.
N. Shroeder, M. Barr.

 

Quantitative Image Analysis

Mouse embryonic stem cells (mESC) and primary fibroblasts (FIB) stained with antibodies against γH2AX and RAD51 proteins (Left). Quantitative analysis of fluorescence foci co-localilocalization before (NoIR) and after X-irradiation (right). 
L. Serrano, J.A. Tischfield.

 

Fluorescence Emission Fingerprinting

hTfR-GFP expression in C. elegans intestine unmixed from intestinal autofluorescence using reference spectra. A) hTfR-GFP; B) intestinal autofluorescence; C) Merge.
C. Chen, C. Rongo.

 

Six colors 3-D FISH of HOX gene clusters and fibrillarin immunostaining (purple) in a neural stem interphase nuclei counterstained by DAPI (blue).
N. Kane-Goldsmith, L. Serrano

 

Tile Scan

Tile- Scan of ten single Z-stack images (20X). Mouse hippocampus stained with ZnT3 (green) and synaptophysin (red) antibodies.
G. Martel, G. Shumyatsky

 

FRAP Analysis

FRAP analysis of Suv39h1-EGFP in NIH3T3 cells. Top) Fluorescence signals from centromeric heterochromatin. The red arrows indicate the 1μm bleached circle. Bottom) Quantification of the halftime of fluorescence recovery and mobile fraction.
N. Kane-Goldsmith, L. Serrano