Bioimaging Systems

Multichannel confocal imaging

The BD CARV II™ imager is suitable for both live cell and fixed cell confocal imaging using a wide variety of fluorophores.

Stack projection of HeLa cells with Qdot® conjugates. Nucleus – Qdot® 655; Golgi – Qdot® 585; Microtubules – Qdot® 525. Quantum Dot Corporation, Hayward, CA.	Stack projection of mouse intestine section labeled with Alexa Fluor® 350 WGA, Alexa Fluor® 568 phalloidin, and SYTOX® Green. Molecular Probes/Invitrogen, Eugene, OR.
3D reconstruction of imaged skin sample labled using pan-neuronal marker, protein gene product 9.5 localized with Cy™3 and basement membrane marker, type IV collagen with CY2. Endothelial cells are stained using Cy™5-Ulex europaeus agglutinin type I. Dr. William R. Kennedy and Gwen Wendelshafer-Crabb, University of Minnesota, Minneapolis, MN.

3D reconstruction

Automation of internal multi-position excitation (8 position), dichroic (5 position) and emission (8 position) filter wheels allows multi-channel 3D and 4D images to be obtained quickly without compromising resolution. Using any of the recommended software packages, 3D and 4D reconstructions can be easily performed.

Maximum projection and orthogonal view created from confocal sections through a sea urchin embryo fixed and stained with anti-tubulin (green) and phalloidin (red). Tubulin (green), Actin (red), (60X 1.4NA) Dr. George von Dassow, Center for Cell Dynamics, Friday Harbor Labs, University of Washington, Friday Harbor, WA.

Orthogonal view (XZ)

Time lapse

Using any recommended imaging software and camera, time lapse imaging of cells or organisms can be performed for long periods of time without significant bleaching or phototoxicity. Depending on the speed of the cellular event being measured, and the installed BD CARV II components, time lapse at a single plane or at multiple planes (4D) can be imaged.

Time lapse images at a single confocal plane of a sand dollar embryo during mid blastula stage (rhodamine-tubulin and Alexa® 488 phalloidin, Molecular Probes, Eugene, Oregon). Dr. George von Dassow, Bill Bement, Center for Cell Dynamics, Friday Harbor Labs, University of Washington, Friday Harbor, WA.

Time lapse images at a single plane of a nematode embryo going through the first cell division stage. GFP-tubulin, GFP-histone. Dr. Fabio Piano, NYU, New York, NY.

High speed calcium imaging

The BD CARV II™ imager, in combination with CCD cameras with on-chip amplification, can be used to image fast fluorescence changes at rates ranging from 50–100 frames per second.

Time lapse recordings at 50 frames per second of calcium sparks in muscle cells loaded with calcium indicator dye Fluo-4, Molecular Probes, Eugene, Oregon. The Roper Cascade 512B CCD camera in conjunction with Metamorph® (Molecular Devices, Sunnyvale, CA) imaging software was used to capture the images.

Fluorescence Recovery After Photobleaching (FRAP)

Our FRAP capability allows software control of a variable rectangular aperture for photobleaching a selected area with full spectrum (white) light. A software protocol can then be set up to record fluorescence before and after bleaching. The recovery kinetics can be analyzed using a variety of commercially available software packages.

Time lapse images and FRAP kinetics of endoplasmic reticulum in a plant cell stained with the membrane dye 3,3-dihexyloxacarbocyanine iodide (DiO6(3), Molecular Probes, Eugene, Oregon). The FRAP protocol including image capture and analysis was performed using BD IPLab™ imaging software.

Dual emission imaging (FRET)

The BD CARV II imager, in combination with the Dual View™ adapter from Optical Insights LLC, Tucson, AZ, allows you to perform dual emission imaging including CFP/YFP and GFP/RFP Fluorescence Resonance Energy Transfer (FRET).