Cytometry is the measurement of number and characteristics of cells. Variables that can be measured by cytometric methods include cell size, cell count, cell morphology (shape and structure), cell cycle phase, DNA content, and the existence or absence of specific proteins on the cell surface or in the cytoplasm. Cytometry is used to characterize and count blood cells in common blood tests such as the complete blood count. In a similar fashion, cytometry is also used in cell biology research and in medical diagnostics to characterize cells in a wide range of applications associated with diseases such as cancer and AIDS.
Image cytometry is the oldest form of cytometry. Image cytometers operate by statically imaging a large number of
cells using optical microscopy. Prior to analysis, cells are commonly stained to enhance contrast or
to detect specific molecules by labeling these with fluorochromes. Traditionally,
cells are viewed within a hemocytometer to aid manual counting.
Since the introduction of the digital camera, in the mid-1990s, the automation level of
image cytometers has steadily increased. This has led to the commercial availability of automated image cytometers, ranging from simple cell counters to sophisticated high-content screening systems.
Flow cytometry
Due to the early difficulties of automating microscopy, the flow cytometer has since
the mid-1950s been the dominating cytometric device.
Flow cytometers operate by aligning single cells using flow techniques. The cells are characterized
optically or by the use of an electrical impedance method called the Coulter principle.
To detect specific molecules when optically characterized, cells are in most cases stained with the same
type of fluorochromes that are used by image cytometers. Flow cytometers generally provide
less data than image cytometers, but have a significantly higher throughput.
Flow cytometry
Cell sorters are flow cytometers capable of sorting cells according to their characteristics.
The sorting is achieved by using technology similar to what is used in inkjet printers.
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The hemocytometer (or haemocytometer) is a counting-chamber device originally designed and usually used for counting blood cells. The hemocytometer was invented by Louis-Charles Malassez and consists of a thick glass microscope slide with a rectangular indentation that creates a precision volume chamber. This chamber is engraved with a laser-etched grid of perpendicular lines. The device is carefully crafted so that the area bounded by the lines is known, and the depth of the chamber is also known.
A Coulter counter is an apparatus for counting and sizing particles suspended in electrolytes. The Coulter counter is the commercial term for the technique known as resistive pulse sensing or electrical zone sensing. The apparatus is based on the Coulter principle named after its inventor, Wallace H. Coulter. A typical Coulter counter has one or more microchannels that separate two chambers containing electrolyte solutions. As fluid that contains particles or cells is drawn through the microchannels, each particle causes a brief change to the electrical resistance of the liquid.
Flow cytometry (FC) is a technique used to detect and measure physical and chemical characteristics of a population of cells or particles. In this process, a sample containing cells or particles is suspended in a fluid and injected into the flow cytometer instrument. The sample is focused to ideally flow one cell at a time through a laser beam, where the light scattered is characteristic to the cells and their components. Cells are often labeled with fluorescent markers so light is absorbed and then emitted in a band of wavelengths.
Immunoengineering is an emerging field where engineering principles are grounded in immunology. This course provides students a broad overview of how engineering approaches can be utilized to study im
This module covers the fundamentals of Flow Cytometry, both practical and theoretical. Students will work in small groups, from sample preparation to data analysis, giving them hands-on time and allow
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