Cellular Investigations Flashcards
1
Q
Define what cellular investigations can help with identification of ;
A
- cell surface phenotype
- intracellular proteins
- cellular function
- secreted products
- abnormal cellular constituents
2
Q
What techniques are typically involved in cellular investigations ?
A
- flow cytometry
- EIA/RIA
- Tissue culture
- PAGE
- genetic (PCR based)
3
Q
Describe flow cytometry
A
- corner stone of cellular analysis
- single cell suspensions analysed - tissues can be digested
- laser light scatter
- fluorescent probe detection - dyes / monoclonal anitbodies for specificity
4
Q
Describe the process of flow cytometry
A
- 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
5
Q
Describe Tissue culture
A
- In Vitro studies of cells require isolation of cells from whole blood
- density gradient centrifugation - fickle/percoll
- immunomagnetic isolation –> paramagnetic beads with monoclonal antibodies
6
Q
What are some kinds of Proliferation Assays?
A
- T cell responses
1. tritiated thymidine
2. mitogenic stimuli - others like flow cytometery
7
Q
What are some examples of soluble mediator assays ?
A
- EIA/RIA (elisa)
- Functional assay
- biomarker assay
8
Q
What are some examples of Apoptosis Assays ?
A
- caspase assay
- ion flux
- cell cycle analysis
- mitochondrial depolarisation
Detect defective apoptosis in;
- ALPS (Autoimmune lymphoproliferative syndrome)
- caspase deficiency
9
Q
Describe BAL & Sputum
A
- Bronchoalveolar Lavage = saline washing of lungs
- Sputum = induced/spontaneous & must processed to release cells
