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Please use this identifier to cite or link to this item: http://hdl.handle.net/1942/8555

Title: A highly reliable and budget-friendly Peltier-cooled camera for biological fluorescence imaging microscopy
Authors: JOLLING, Koen
VAN DE VEN, Martin
Issue Date: 2007
Citation: JOURNAL OF MICROSCOPY-OXFORD, 228(3). p. 264-271
Abstract: The SAC8.5, a low-cost Peltier-cooled black and white 8-bit CCD camera for astronomy, was evaluated for its use in imaging microscopy. Two camera-microscope configurations were used: an epifluorescence microscope (Nikon Eclipse TE2000-U) and a bottom port laser scanning confocal microscope system (Zeiss LSCM 510 META). Main advantages of the CCD camera over the currently used photomultiplier detection in the scanning setup are fast image capturing, stable background, an improved signal-to-noise ratio and good linearity. Based on DAPI-labelled Chinese Hamster Ovarian cells, the signal-to-noise ratio was estimated to be 4 times higher with respect to the currently used confocal photomultiplier detector. A linear relationship between the fluorescence signal and the FITC-inulin concentrations ranging from 0.05 to 1.8 mg mL(-1) could be established. With the SAC8.5 CCD camera and using DAPI, calcein-AM and propidium iodide we could also distinguish between viable, apoptotic and necrotic cells: exposure to CdCl2 caused necrosis in A6 cells. Additional examples include the observation of wire-like mitochondrial networks in Mito Tracker Green-loaded Madin-Darby canine kidney cells. Furthermore, it is straightforward to interface the SAC8.5 with automated shutters to prevent rapid fluorophore photobleaching via easy to use ASTROVIDEO software. In this study, we demonstrate that the SAC8.5 black and white CCD camera is an easy-to-implement and cost-conscious addition to quantitative fluorescence microfluorimetry on living tissues and is suitable for teaching laboratories.
Notes: Hasselt Univ, Ctr Environm Sci, Diepenbeek, Belgium. Hasselt Univ, Biomed Res Inst, Physiol Lab, Diepenbeek, Belgium.
URI: http://hdl.handle.net/1942/8555
ISI #: 000251251400002
ISSN: 0022-2720
Category: A1
Type: Journal Contribution
Validation: ecoom, 2008
Appears in Collections: Research publications

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