– Cell Biology Laboratory

The Cell Biology Laboratory is equipped with the Leica TCS SP5 II confocal microscope, Nikon Eclipse 80i Advanced research digital microscope and the BD FACS Aria II Flow cytometer.

Leica TCS SP5 II confocal microscope

The confocal microscope offers several advantages over conventional optical microscopy, including controllable depth of field, the elimination of image degrading out-of –focus information, the ability to collect serial optical sections from thick specimens. The key to the confocal approach is the use of spatial filtering to eliminate out-of-focus light or flare in specimens that are thicker than the plane focus. There has been tremendous explosion in the popularity of confocal microscopy in recent years, due in part to the relative ease with which extremely high-quality images can be obtained from specimens prepared for conventional optical microscopy. There are applications in many areas of research in cell and molecular biology. Our lab is designed with dim lighting to reduce fading of the fluorescence probe during imaging. The confocal microscope also comes with an incubation chamber to allow real-time live imaging of cells for more than 24 hours.

Nikon Eclipse 80i Advanced Research Digital Microscope

The Nikon microscope delivers remarkably high signal to noise ratios in producing fluorescence images. Fluorescent-tagged structures or proteins can be located and the cellular morphology of specimens can be visualized using a high-performance DIC method in combination with Hi S/N epi-fluorescence illumination. The microscope is for the visualization tissue/cells fixed on glass slide.

BD FACS Aria II Flow cytometer

The FACSAria II from Becton Dickinson (BD Biocsiences, San Jose, USA) is a high speed cell sorter for measuring and sorting fluorescently labelled cells. These cells can be prokaryotic (e.g. bacteria) or eukaryotic (e.g. mammalian) with a minimum size of 0.2 um (maximum 100 um). Fluorescence staining is usually performed with antibodies specific to the proteins of interest, which can be intra-cellular or on the cell surface. These primary antibodies are conjugated to a fluorescent dye, or a fluorescent secondary antibody may be used. Fluorescent substrates or compounds can also be detected.

A laser excites the fluorescent dyes bound to the cells. The emitted light has a longer wavelength and passes through a bandpass filter to shut out the excitatory and other unwanted light. The strength of this fluorescent light is measured with a detector (PMT) for each cell and is displayed in form of a histogram.