The Lakkaraju lab uses the Andor Revolution XD microscopy system
(with high speed spinning-disk confocal, TIRF, and Mosaic capabilities) to study intracellular organelle trafficking (endosomes, lysosomes and autophagosomes) and follow endo- and exocytic events at the cell membrane in real time. This is a shared instrumentation facility supported by the Department of Ophthalmology & Visual Sciences, the School of Medicine and Public Health and by grants from the Research to Prevent Blindness Foundation and Reeves Foundation for Macular Degeneration Research.
Our objective is to discover the cellular basis of blinding diseases such as age-related macular degeneration (AMD), which affects over 30 million people worldwide currently. The long-term goal of our research is to use this knowledge to develop novel therapies that could help prevent devastating vision loss. Our research also seeks to understand whether mechanisms that underlie age-related macular degeneration share features with other diseases of aging such as Alzheimer’s disease, Parkinson’s disease, atherosclerosis, etc. A common thread in these seemingly disparate diseases is abnormal cholesterol metabolism and disrupted lysosome-autophagic function.
Andor Revolution XD Confocal inverted microscope
- Nikon Objectives:
- 100x Apo TIRF (Oil, NA = 1.49, WD = 0.12 mm) with Temperature Correction collar (for 23°C and 37°C)
- 60x Plan Apo VC (Oil, NA = 1.4, WD = 0.13 mm)
- 40x Plan Fluor (Oil, NA = 1.3, WD = 0.2 mm)
- 20x Plan Fluor (Dry, NA = 0.5, WD = 2.1 mm)
- Andor Laser Combiner (ALC): Four solid state lasers at 405, 488, 561, and 640 nm with 10 millisecond switch time and active blanking control to minimize unnecessary illumination.
- Confocal emission bandpass filters:
- DAPI: 447/60 (417-477nm)
- GFP: 525/30 (510-540nm)
- RFP: 607/36 (589-625nm)
- Far-red (CY5): 685/40 (665-705nm)
- Okolab CO2 cage incubator: Environmental regulation of temperature, CO2, and humidity to achieve cell culture conditions. Temperature is thermostat-regulated to ± 0.1°C from 3° above ambient temperature up to 50°C.
Supported by the Department of Ophthalmology and Visual Sciences, the University of Wisconsin School of Medicine & Public Health, the Carl and Mildred Reeves Foundation for Macular Degeneration Research, and generous funding provided by the Schuette/Kraemer Family gift.