Cellular Investigations

Question 1

Define what cellular investigations can help with identification of ;

Answer 1

• cell surface phenotype
• intracellular proteins
• cellular function
• secreted products
• abnormal cellular constituents

Question 2

What techniques are typically involved in cellular investigations ?

Answer 2

• flow cytometry
• EIA/RIA
• Tissue culture
• PAGE
• genetic (PCR based)

Question 3

Describe flow cytometry

Answer 3

• corner stone of cellular analysis
• single cell suspensions analysed - tissues can be digested
• laser light scatter
• fluorescent probe detection - dyes / monoclonal anitbodies for specificity

Question 4

Describe the process of flow cytometry

Answer 4

• cells move in a single file through laser
• laser light is scattered depending on the size of the cell & on the complexity of the cell contents
• fluorescent dyes & antibodies can be used to reveal further info about cells
• some machines permit specifically labelled cells to be collected for further analysis

Question 5

Describe Tissue culture

Answer 5

• In Vitro studies of cells require isolation of cells from whole blood
• density gradient centrifugation - fickle/percoll
• immunomagnetic isolation –> paramagnetic beads with monoclonal antibodies