DeltaVision Elite

System Overview

The Imaging Core's new DeltaVision Elite Deconvolution/TIRF microscope system (installed on March 24-25, 2014) is a 3-D deconvolution (image restoration) system with 488nm and 561nm laser lines for TIRF (total internal reflection microscopy).

Deconvolution uses the properties of the microscope system (point spread function) to deblur and remap out-of-focus fluorescent signal in order to enhance image resolution and contrast. TIRF is a technique used to image samples within ~100-200nm of the coverslip surface. 

The system is equipped 60x oil, water and TIRF objective lenses, a CMOS camera, and DAPI, CFP, GFP, YFP, TRITC, mCherry, and CY5 fluorescence filters. It is recommended for fluorescence imaging of thin fixed specimens and time-lapse imaging of live cells. The proprietary solid state illumination system provides uniform, high intensity light to the sample, enabling detection of small, dim objects, and allowing for shorter exposure times and fast image acqusition. An incubation system with temperature control and 5% CO2 is available for long-term live cell imaging experiments. The system is equipped with an Ultimate Focus laser that is capable of continually monitoring the position of the stage to eliminate z-drift during time-lapse studies.  

When in TIRF mode, the system images samples within 100-200nm of the coverslip surface. The TIRF angle is variable and each wavelength is chromatically corrected to ensure all laser lines penetrate the sample to the same depth.

New Users

Faculty and students who wish to use the microscope should follow the instructions in the New Users Guide.

Prepared slides will be used during the training session in order to acquaint new users with all aspects of the microscope, however, new users are also encouraged to bring their own samples to the training session if any are available.  It is also highly recommended that new users review the DeltaVision Imaging System Manual and/or the DeltaVision Short Imaging Guide before training.

Rate Schedule

Following January 1, 2015, the full rate schedule applies. Fees are used to help cover the cost of the service contract on the microscope, which is partially subsidized by the department, and to help pay for materials such as lens paper and lens cleaning solution. 

Policy

The microscope is available to trained users on an equal basis. An individual user may reserve one four hour block of time during peak hours and an additional 6 hours of time during off-peak hours. Users who need more time for long-term live cell imaging experiments should contact Amy Beaven. Scheduling is accomplished using Google Calendar.  For information on how to apply for and schedule instrument time using Google Calendar, please click here.  

All users are encouraged to familiarize themselves with the complete list of policies governing the use of the facility.

References

The facility manager is available during normal business hours to provide technical assistance to users.  Additional resources include the following Imaging Core protocols:

Other Useful Links:

System Specifications

Microscope:

  • Olympus inverted IX-71 base
  • Precise motorized X,Y,Z stage with 25mm x 50mm travel area
  • Clear environmental chamber with regulated temperature, CO2 and humidity
  • Ultimate focus (hardware-based focus stability) 
  • Long working distance DIC condenser lens

Objective lenses:

  • 60x, 1.2NA UPLSAPO water lens (WD 0.28mm)
  • 60x, 1.49 NA oil APON TIRF lens (WD 0.1mm)
  • 60x, 1.42 NA PlanApo oil lens

Insight SSI 7-color solid state fluorescence illumination with the following excitation filters:

  • DAPI (390/18)
  • CFP (438/24)
  • GFP/FITC (475/28)
  • YFP (513/17)
  • TRITC (542/27)
  • mCherry/AF594 (575/25)
  • CY5 (632/22)

Filter cubes/dichroics/emission filters (click here for a chart of available filter combinations):

  • Polychroic beam splitter suitable for DAPI, FITC, RD-TR-PE, CY5
  • Poychroic beam splitter suitable for DAPI, FITC, Alexa 594, CY5 and GFP/mCherry
  • Polychroic beam splitter suitable for CFP/YFP/mCherry
  • Single band pass emission filters for DAPI (435/48), CFP (475/24), GFP/FITC (525/48), YFP (548/22), TRITC (597/45), mCherry (625/45), CY5 (679/34).

Camera:

  • DV Elite CMOS camera

Contact Information

Amy Beaven
Director
Imaging Core Facility
0107 Microbiology Building
University of Maryland
abeaven@umd.edu
301-405-7238

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