History

The CSIF purchased the DeltaVision deconvolution microscope system from Applied Precision, Inc. in June 1997. The DeltaVision includes a wide-field microscope and software to deconvolve the raw data captured on the cooled CCD camera. The system is based on the one developed in the laboratories of Drs. John Sedat and David Agard at UCSF. (see publication list for Sedat lab)

The system was upgraded to a RT system in 2006. The cooled CCD camera was upgraded to a newer interline CCD camera with a peak QE of 60% (CoolSNAP HQ, Princton Instruments). Download a copy of the camera specifications here. Live cell fitler sets designed specifically for CFP/YFP fret imaging as well as eGFP/dsRED live cell imaging were also added. Other features include new SoftWoRx 3.4.5 software running on a Linux workstation and SoftWoRx Explorer Suite running on a Windows workstation, which includes Bitplane's Imaris software.

A brief explanation and references for deconvolution microscopy can be found at Molecular Expressions' Optical Microscopy site and more detailed info at SVI's Wiki pages at SVI.nl/wiki

In addition to the Delta Vision deconvolution software, the CSIF has installed on the computer St. Francis SVI's Huygens Essential deconvolution software that can perform image restortaion on widefield, confocal and 2P datasets.

System specifications

Microscope - IX-70 inverted microscope with epi-fluorescence attachment.

Objectives -

Name	Magnification	NA	Immersion	working distance
UAPO /340	20x	0.75	air	0.55 mm
UPLAN APO	40x	0.85	air	0.20 mm
U-APO	40x	0.65-1.35	oil	0.10 mm
PLAN APO	60x	1.40	oil	0.10 mm
UPLAN APO	60x	1.20	water	0.25 mm
UPLAN APO	100x	1.40	oil	0.10 mm

Light source - Excitation light is provided by a mercury arc lamp which provides illumination from the UV to the near-IR. This system can excite DAPI and Hoescht, eCFP FITC, eGFP, eYFP, Rhodamine, Cy3, Texas Red, Cy-5, dsRed (and some variants) and most Alexa dyes.

Excitation filters available:

• 360/40 for DAPI, Hoescht
• 405/30 for Lucifer yellow
• 490/20 for FITC, GFP, PE
• 555/28 for Rhod, TR, Cy3
• 640/20 for Cy5
• 340/14 for Fura2 (denominator)
• 380/14 for BFP, Fura2 (numerator)
• 436/10 for CFP
• 500/20 for YFP

Emission filters available:

• 457/50 for DAPI, Hoescht
• 528/38 for FITC, GFP, PE, Fura-2, Lucifer yellow
• 617/73 for Rhod, TR, Cy3
• 685/40 for Cy5
• 470/30 for CFP
• 535/30 for YFP

Detection:

• A Princton HQ CoolSNAP interline cooled CCD camera is used for both high resolution imaging of fixed samples as well as fast live cell imaging.
  
• Triple labelled specimens can be imaged at all three wavelengths.

Software: SoftWoRx, version 3.4.5

• controls image acquisition parameters
• simple interface for automated deconvolution process
• produces three-dimensional reconstruction of datasets
• includes modelling tools (wire-frame,solid surface)
• contains limited analysis tools for numerical computations.

• Enkil2 (Linux OS, microscope workstation)
• Memnoch (SGI Octane workstation, SoftWoRx 2.5) and Memnoch2 (Windows 64bit, dual processor, 8GB RAM, Bitplane Imaris, Huygens Essential)
  

Manuals

To obtain a copy of the CSIF user manuals for this system, you will need to have Adobe Acrobat reader installed on your computer.

Download CSIF user manual - Applied Precision SoftWoRx v3.4.5 RT upgrade


Download CSIF DeltaVision server information sheet - explains where data is stored, how long it is stored, and how to ftp from the CSIF computers to your computer.