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DeltaVision Elite Deconvolution/TIRF

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  • Recommended for imaging thin, fixed specimens and time-lapse imaging of "small, dim, live" cells.
  • Illumination: choice of fluorescence filters (DAPI, GFP, dsRed, mCherry, Cy5, CFP, YFP) for widefield and deconvolution (see details below), and two TIRF lasers: 488, 561nm.
  • Objectives: 10x, 20x, 40x dry, 63x oil, 63x water, 63x TIRF
  • Features: sCMOS camera, motorized x-y stage, "big box" incubation with temperature, CO2 and humidity control.

New Users

Faculty and students who wish to use the microscope should follow the instructions in the New Users Guide. Training takes ~2 to 3 hrs, with additional mandatory follow-up sessions required until the director determines the user is capable of operating the microscope independently. Users are encouraged to bring their own samples to the training session. If none are available, prepared slides will be used. It is also highly recommended to review the DeltaVision Quickstart Guide before training begins.

Policy

The microscope is available to trained users on an equal basis. Individual users may reserve one four hour block of time during "peak" daytime hours, and an additional 6 "off-peak" hours (all other times). Investigators performing live cell imaging experiments may signup for 12 hours blocks of time after 6pm.

For a comprehensive list of policies (including BSL-2 protocols), please see the Facility SOP.

Rates

The rate schedule applies to all users and is current for the 2022/2023 academic year. 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, lens cleaning solution and mercury bulbs.

System Details

The  DeltaVision Elite Deconvolution/TIRF microscope system 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  a number of objective lenses (including 60x oil, 60x water and 60x TIRF), 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 acquisition. 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 can help prevent z-drift during time-lapse studies.  

When in TIRF mode (excitation 488, 561nm), the system images samples within 100-200nm of the coverslip surface. The TIRF angle is variable and each wavelength is chromatically corrected to ensure both laser lines penetrate the sample to the same depth.
Microscope Stand
Olympus IX71 Inverted
Illumination
TIRF: 488, 561nm lasers. Widefield/deconvolution:  7 color InsightSSI solid state fluoescence illumination with the following filters: DAPI (390/18), CFP (438/24), GFP (475/28), YFP (513/17), TRITC (542/27), mCherry (575/25), Cy5 (632/22)
Polychroics and Emission
Polychroic beamsplitters: (1) DAPI/FITC/TRITC/Cy5  (2) DAPI/FITC/mCherry/Cy5  (3) CFP/YFP/mCherry. Emission filters: DAPI (435/48), CFP (475/24), GFP (525/48), YFP (548/22), TRITC (597/45), mCherry (625/45), Cy5 (679/34). Click here for a summary chart of beamsplitters and filters.
Light detector
Front illuminated sCMOS, 2560 x 2160 pixels, Pixel size, 6.45 μm, Readout speed, 95 MHz, 286 Mhz.
Objective Lenses
10x, 20x 0.75NA, 40x 0.6NA, 63x 1.42NA oil, 63x 1.2NA water, 63x 1.49NA TIRF
Incubation
"Big Box" incubator with temperature, humidity, and CO2 control
X-Y Stage
Precise motorized X,Y,Z stage with 25mm x 50mm travel area
Software
SoftWorx

References

Imaging Core protocols:
  • DeltaVision Start-Up and Shut-down Procedure
  • DeltaVision Policy
  • DeltaVision Quickstart Guide
  • DeltaVision Filter Guide

Contact Us

Amy Beaven
Facility Director
abeaven@umd.edu
301.405.7238
0107 Microbiology Building

Dr. Charles Delwiche
Faculty Supervisor
delwiche@umd.edu
301.405.8286
2108 Bioscience Research Building

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  • Home
  • Imaging
    • Imaging Core >
      • Instruments >
        • Zeiss LSM 980
        • Leica SP5X
        • Zeiss LSM710
        • DeltaVision
        • Nikon Eclipse
        • Zeiss AxioObserver
      • NEW USER GUIDE
      • RATES
      • FAQ
      • IMAGE GALLERY
    • Imaging Incubator >
      • INSTRUMENTS >
        • Zeiss LLS7
        • PerkinElmer
        • ASI diSPIM
        • JPK AFM
        • 3P
      • Rates
      • NEW USER GUIDE
  • Flow Cytometry
  • Genomics