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Leica DMI4000 with Yokagawa CSU-X1 Spinning Disk Confocal Attachment


Description

The Leica DMI4000 Microscope with Yokagawa CSU-X1 Spinning Disk Confocal Attachment offers a full environmental enclosure, programmable motorized stage, and sensitive EM-CCD camera make this spinning disk confocal system ideal for long-term multipoint timelapse microscopy.  It is also excellent for large-scale automated tile acquisition and image stitching.


Applications

  • Long-term multi-point timelapse microscopy
  • Fluorescence, brightfield, differential interference contrast (DIC), and polarization microscopy
  • Large-scale image tile acquisition and stitching

Leica DMI4000 Spinning Disk Confocal

Spinning disk microscope
Technical Specifications: Spinning Disk Tech Sheet (PDF)

Scheduling Core Instruments

Registered users can access the PVIC online scheduler to reserve and use the Penn Vet Imaging Core instruments.

If you are not already registered, please contact the core manager.

Application Examples

Multipoint Timelapse on Spinning Disk

muti-point timelapse, spinning disk

A timelapse experiment done on the Penn Vet Imaging Core’s Yokagawa spinning disk confocal microscope is shown schematically.  Multiple stage positions, different imaging modes (such as brightfield and laser confocal of GFP fluorescence in this example), number and range of depth for multiple focal planes, and time interval and duration can all be specified within the MetaMorph software to automate image acquisition.  A full environmental enclosure around the microscope allows cells to be maintained at a stable 37°C temperature and 5% CO2 level for long-term imaging. 

Images of HEK293T cells transfected to express GFP-tagged ebola VP40 protein courtesy of Gordon Ruthel, Xiaohong Liu, Bruce Freedman, and Ron Harty, Department of Pathobiology, UPenn School of Veterinary Medicine.


VP-40 Timelapse Movie


This timelapse movie of HEK293T cells that were transfected to express a GFP-tagged ebola VP40 protein was acquired on the Penn Vet Imaging Core’s Yokagawa spinning disk confocal microscope.  The images were rendered as color heatmaps of GFP fluorescence intensity with MetaMorph imaging software.  Elapsed time is shown as hours:minutes:seconds in the lower left corner. 

Movie courtesy of Gordon Ruthel, Xiaohong Liu, Bruce Freedman, and Ron Harty, Department of Pathobiology, UPenn School of Veterinary Medicine.

Large-scale image tile acquisition and stitching

CEACGN OD vasculature, Imaging Core

The vasculature pattern of a dog retina pictured here was pieced together from 1600 individual image fields acquired with a 10x objective lens and 488 nm laser on the Penn Vet Imaging Core’s Yokagawa spinning disk confocal microscope.

For each field, an image Z-stack was taken at 5 µm intervals over an almost 100 µm thickness and the maximal projection image for each field was used for the montage.

The images were stitched together with MetaMorph imaging software, which was also used to automate the acquisition of the 1600 individual images. The resulting fluorescence image was color-inverted for the view presented here.

Image courtesy of William Beltran, Section of Ophthalmology, Department of Clinical Studies, UPenn School of Veterinary Medicine.