The document discusses various technologies used at the House Ear Institute including genomics, proteomics, and imaging. It describes how researchers are using these tools to study diseases like neurofibromatosis type 2 (NF2) at the molecular level in order to develop personalized treatments and therapies. Maintaining high quality biospecimens is important for enabling various types of research.

1. HOUSE EAR INSTITUTE Robert Gellibolian, Ph.D.

4. Spatial Dynamic Range: ~10 12 ~ 10 -12 m ~ 10 -9 m ~ 10 -6 m ~ 1 m ~ 10 -2 m ~ 10 -1 m Temporal Dynamic Range: ~10 15 ~ 10 -6 s ~ 10 -3 s ~ 1 s ~ 10 3 s ~ 10 6 s ~ 10 9 s Molecular Events Diffusion (Cell Signaling) Motility Cell Division Protein Turnover Human Lifetime ORGANISM ORGAN ORGAN SYSTEM TISSUE CELLS MOLECULES ATOM / BOND

20. IONIZER MASS ANALYZER DETECTOR MALDI Electro-Spray Ionization (ESI) Time-Of-Flight (TOF) Quadrapole Ion-Trap Electron Multiplier (EM) Anatomy of a Mass Spectrometer Sample + _

21. What we have to do this: Database Search & Probability Match Eksigent 2D nano-LC Separation of molecules ABI 4000 QTRAP Determine “mass” of molecules

25. 1951: “The Man in the White Suit” starring Alex Guinness 1981: “Blade Runner” starring Harrison Ford

27. LM EM 0.3 microns

29. SEM TEM

31. Example: Zebrafish

32. Mammalian Inner Ear Glen MacDonald Hair Cells Dr. Sonja Pyott

33. ZEISS LSM 710 w/ CONFOCOR 3 LEICA SP5

36. Biorepositories Imaging Chemical & Molecular Therapies Cell & Tissue Therapies Immune & Monitoring Vaccines Translational Research & Resources Basic Research Prototype Design or Discovery Preclinical Development Clinical Development FDA filing/Approval & Launch Preparation Translational Research

37. Powerful Tools : Powerful Risks Low Quality Samples Low Quality Data GIGO

39. Finding the targets for detection, therapy, prevention Genomics Proteomics Metabolomics Identification of targets for drug development, treatment and prevention Identify biologic variations that determine drug efficacy and drug toxicity Defining markers for susceptibility Validation of new therapeutics ALL DEPEND ON HIGH QUALITY BIOSPECIMENS

40. Consensus of the Broad Scientific Community: The lack of high-quality human specimens has become the limiting factor for post-genomic biomedical science.

42. Patient The “specimen” exists in situ within the specific biologic context of the patient

43. Medical/Surgical Procedures Examples include administration of antibiotics, anesthesia and other drugs. At this point, anesthesiologists, nurses, and doctors may introduce variables into the procedure that change the biologic context of the “specimen” and, as a result, cause dynamic changes in the molecular profiles within the “specimen”.

44. Acquisition Various methods of acquiring the “specimen” may introduce specific types of stress or trauma to the living specimen. Examples include time delays (from handling to delivery), and packaging (changing exposure to desiccation or oxidation).

45. Handling/Processing Preparation variables introduced prior and during biologic stabilization of the specimen include: time @ RT, RT, type and time in fixative, method & rate of freezing, size of specimen aliquots. Human involvement contribute to this process.

46. Storage Variables include storage temperature, duration, progressive dehydration, desiccation and oxidation may contribute to this process.

47. Distribution Variation sin transport conditions may lead to alterations in the specimen. Package, transport and receiving people all contribute to this type of variations.

48. Scientific Analysis Different methods of analysis for various classes of biomolecules may be affected by any of the factors already mentioned. Even with a single analytical method, each individual investigator and technician involved in the specimen analysis may contribute to such variation.

49. Restocking Environmental variables may be introduced through the return of unused samples from research laboratories to storage.

50. CLINICAL STUDIES SECTIONS Cell Struct & Function Developmental Biology NEURAL TUMOR RESEARCH (NF2) SECTIONS Pathogenesis Cell Differentiation CORE TECHNOLOGIES GENOMICS PROTEOMICS IMAGING TISSUE BANKING