1. Laboratory toxicology

2. Toxicological methods In vitro –Cell cultures –Cell-free systems –Mechanistic In vivo (lab animals) –Acute –Subchronic –Chronic –Reproductive

3. In vitro toxicology tests What is the chemical (or other hazard)? What is the appropriate cell type? What is the outcome we expect to see as effect? What is the dose range to test? What are the detection methods? Sensitivity of detection (#molecules change)? Replicates and controls

4. Detecting DNA breaks the DNA ladder

5. Comparing samples for DNA breaks on gel

6. Detecting apoptotic cells

7. ELISA kit for apoptotic cells (Roche)

8. The dose-effect result: compare test and control

9. (Immuno)fluorescence detection method

10. Flow cytometry

11. Western blotting

12. Microarrays in gene-expression toxicology Used to identify patterns of gene expression In response to treatment with xenobiotic

13. Basic in vivo toxicity studies Type of studyNumber of exposuresOutcomes AcuteSingle Determine lethal dose; Identify potential target organs; Indicate comparative toxicity SubchronicIntermittent Determine specific target organs Repeated that may be damaged at levels (up to 90 days) below which adverse effects are observed ChronicIntermittent Identify specific abnormalities Repeated or diseases, determine the over long periods conditions of exposure and dose (up to lifetime) required for specific forms of damage

20. Sample size and sensitivity of detection

24. Immunohistochemistry for animal tissues

25. The cost of testing

26. Alternatives to animal testing