This document discusses various cellular techniques used for separating, analyzing, and studying cells, including: 1. Fluorescence-activated cell sorting (FACS) which uses fluorescent antibodies to separate and analyze cells by flow cytometry. 2. Immunofluorescence techniques like direct testing with primary antibodies, indirect testing with secondary antibodies, and sandwich assays using capture and secondary antibodies. 3. Microscopy techniques like light, electron, and fluorescence microscopy used to visualize cells. 4. Functional assays like the chromium release assay to measure a cell's ability to kill target cells and limiting dilution analysis to estimate the frequency of precursor cells.

1. Cellular techniques G. Groenhof s0729884 S. Hemelaar s0729906 S. Hofstraat s0729914 K. Jenniskens s0729949 Y. Khaled s0729981 L. De Kroon s0730041 S. Van Kuijk s0625337

3. Intrinsic fluorescence

4. Buoyant density

5. Adherence

8. Immunofluorescence techniques Direct test Primary antibodies Indirect Secondary antibodies Sandwich Capture antibodies Secondary antibodies

9. Microscopy Light microscopy Histochemical colouring Electron microscopy Electron-dense immunolabel (colloidal gold) UV microscopy Fluorescence

10. Chromium release assay Ability of CTL to kill target cells (functional) Target cell labeled with Chromium 59 (intracellular) Cytotoxicity: release of radioactive protein into medium Repetition at different ratios Effector cells vs. Target cells

11. Limiting dilution analysis Estimate frequency of precursor cells Precursor cells: indication of immune response Cell suspension (~1 cell/ sample) Poisson distribution: 37% of samples contain no precursor cells

12. Plaque technique Agar gel coated with RBC Add complement B cells added Antibodies produced Lysis of RBC Clear spots on agar B cells in clear spots

13. ELISpot Immobilized antigen on well plate B cells added Produced antibodies bind to antigen Secondary antibody added Labeled with enzyme Coloured product solid in gel

14. Stadia of the cell cycle Interphase: G1 phase: increase cytoplasm, production proteins S phase: DNA replication G2 phase: production particles for mitosis Mitosis G0 phase: resting cell

15. Cell proliferation Properties for measuring Mitotic activity DNA activity Metabolic activity Cell proliferation measuring techniques Direct counting of cells Measuring 3H-thymidine, adding cytokines Scintillation counter Detecting dehydrogenase activity increase ELISA

17. 3 different antigens in 4 different concentrations

18. 3 different kinds of antibiotics