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Current Research

Microscope Imaging Facility


The Microscopic Imaging Facility at The Aaron Diamond AIDS Research Center supplies facilities for live-cell deconvolution microscopy. The facility houses an Applied Precisions DeltaVision restoration microscope, utilizing an Olympus IX-70 inverted microscope platform. This microscope system is contained in a BSL-3 facility for work with infectious material. A Bioptechs Focht Live-Cell Chamber system (FCS2) is available for continuous observation of living cells on the DeltaVision microscope. Oil and water immersion objectives are available for work in various cell culture conditions.

DeltaVision Microscope


DeltaVision available optical filters
Fluorochrome Excitation (nm) Emission (nm)
DAPI, Hoechst 360/40 457/50
Alexa Fluor 405 405/30  
CFP 436/10 470/30
FITC, GFP 490/20 538/38
YFP 500/20 535/30
PE, Tx Red, Rhodamine,
PKH26, Propidium Iodide
555/28 617/73
Alexa Fluor 568 575/25  
Alexa Fluor 594, 7AAD, Nile Red   630/40
APC, Cy5 640/20 685/40


DeltaVision objective lenses
Lens ID# Lens Type Magnification/NA Coupling Media Working Distance (mm) Depth of Field (um) Comment
10107 UPlanApo 10x/0.40 Air 3.1 3.16  
10205 UApo/340 20x/0.75 Air 0.55 0.70  
10404 UPlanApo 40x/0.85 Air 0.20 0.56  
10403 UApo/340 40x/1.35 Oil 0.10 0.32 0.65-1.35 NA
10602 PlanApo 60x/1.40 Oil 0.10 0.28  
10603 UPlanApo 60x/1.20 Water 0.25 0.36  
10003 UPlanApo 100x/1.35 Oil 0.10 0.32 0.50-1.35 NA


Image Gallery

Click to view a large version
GFP-Vpr labelled HIV-1 virus particles (Green) bound to the surface of target cells labeled with rhodamine-concanavalinA (red) and Hoechst 33258 (blue)
Image by Paul Bieniasz
and Theodora Hatziioannou

DeltaVision Primer
The following illustrates the technique of deconvolution and restoration microscopy as implemented on the API DeltaVision system.

This image was acquired from a DAPI-stained preparation of 293T cells, provided by Dr. Lubbertus Mulder at ADARC, acquired at 100X magnification.

The DeltaVision microscope is based upon a standard fluorescence microscope, with the addition of precision X, Y and Z stage control. After finding and focusing on a field of interest, the DeltaVision system is set up by the investigator to automatically acquire a number of images (image stack) while the system mechanically focuses through the cell at precise intervals. The following image is number 16 from an image stack of 20 images, each of which is an image taken as the focus was changed by 0.2 um.
At this point the image-processing computer takes over, using a Point Spread Function (PSF) file as a reference to deblur or deconvolve the original image stack. The PSF is a file that characterizes the microscope’s optical characteristics using the same objective lens, coverslip and immersion oil.

This operation generates another image stack of deconvolved images, of which this is again the 16th image from the stack of 20. Notice the increase in resolution obtained by the removal of blurring.

This image stack may be further computer processed to yield a 3-D image of the cells.

Nikon Eclipse TS300 inverted microscope
Filter sets for GFP+DsRed or CFP+YFP, a Spot Insight QE color digital camera for brightfield and fluorescence imaging.

Other Equipment


Molecular Dynamics Store 840 Imaging Systems
The Store imaging system is an optical scanner that produces digital images of Labeled Samples. The Store 840 system includes the phosphor screen and the chemifluorescence/blue-excited fluorescence virtual instruments.

Applied Biosystems Real Time PCR
The Applied Biosystems Real Time PCR system (7500 system) uses fluorescent-based PCR chemistries to provide quantitative detection of nucleic acid sequence using real-time analysis and qualitative detection of nucleic acid sequence using end-point and dissociation curve analysis. The 7500 system allows you to perform several assay types with plates or tubes in the 96-well format.
Thermo Scientific NanoDrop1000
The Thermo Scientific NanoDrop1000 Spectrophotometer enables highly accurate UV/Vis analyses of 1 ul samples. The sample retention system eliminates the need for cuvettes and capillaries which decreases the measurement cycle time. In addition, the high absorbance capability eliminates the need for most dilutions.