Microscopy
Confocal Microscopes
Lighhouse currently operates three spectral confocal microscopes:
- Zeiss LSM 880 with AiryScan
- Zeiss LSM 710
- Leica TCS SP2 AOBS
Confocal microscopes can take images in which only the focal plane of a sample is visible. This results in an improved signal to noise ratio, as well as an increase in effective resolution relative to those made with a standard fluorescence microscope. Confocal microscopy is of potential interest in a wide range of experiments, for example those involving thick specimens, intracellular studies, and colocalization, as well as in studies where the three-dimensional structure of the sample is important.
All of the systems are spectral confocal microscopes, and the lasers available on the different systems are listed below.
Laser Wavelength(s) | Examples of Dyes or Proteins Which Can Be Used |
Violet 405nm | DAPI, Alexa 405, Pacific Blue, Brilliant Violets (421, etc.) |
Multiline Argon (458 nm, 488 nm, 514nm) | Sytox Blue, ECFP, Spectrum Aqua, Brilliant Violet 480 Alexa 488, FITC, CY2, EGFP, YOYO-1, DiOC6(3), JC-1 EYFP, TOTO-1, Oregon Green 514, Rhod-123, Bodipy-FL |
543 nm (Leica) 561 nm (Zeiss 710/880) 594 nm (Leica) | CY3, TRITC (Rhodamine), Alexa 546, Alexa 555, Propidium Iodide, DsRed,mCherry, Texas Red, Alexa 594 |
Red 633 nm | CY5, Alexa 633 / 647, TOTO-3, DRAQ-5, DRAQ-7 |
Both Zeiss microscopes are equipped with climate control chambers for live cell imaging/timelapse measurements. The LSM 880 with AiryScan is a super resolution microscope which is able to take images of specimens beyond the diffraction limit.
Only those investigators who have been previously trained to use the microscopes by the Core Facility will be allowed to use them without an operator. For information about reserving time on the instruments, or about labeling strategies for your experiments, just contact the facility.
Lighthouse also has separate computer workstations with deconvolution, 3D reconstruction and image analysis programs which can be used to help analyze images and Z-stacks taken with the microscopes. For freeware to help you with examination of your images please see our confocal links of interest page below.