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Tesi etd-03092024-121257


Tipo di tesi
Tesi di laurea magistrale
Autore
PARENTI, VITTORIO
Indirizzo email
v.parenti2@studenti.unipi.it, vittorioparenti98@gmail.com
URN
etd-03092024-121257
Titolo
A QUANTITATIVE METHOD BY SUPER-RESOLUTION MICROSCOPY TO CHARACTERIZE EARLY DNA-DAMAGE AND REPAIR INDUCED BY FLASH RADIOTHERAPY
Dipartimento
FISICA
Corso di studi
FISICA
Relatori
relatore Prof.ssa Cella Zanacchi, Francesca
correlatore Prof. Capaccioli, Simone
Parole chiave
  • Fluorescence Microscopy
  • DNA-damage
  • Radiotherapy
  • Biophysics
  • Super-resolution Microscopy
Data inizio appello
25/03/2024
Consultabilità
Non consultabile
Data di rilascio
25/03/2027
Riassunto
My research explores cutting-edge radiotherapy techniques, particularly FLASH radiotherapy, which delivers high doses rapidly with minimal damage to healthy tissues. I employed super-resolution microscopy, specifically the STORM (STochastic Optical Resolution Microscopy) technique, which offers an effective resolution below 20 nm, to quantitatively analyze the effects of irradiation on DNA damage and repair proteins. Additionally, I optimized a quantitative method based on clustering analysis for gammaH2AX histones characterization, fundamental proteins traditionally linked to DNA damage and repair. As a case study, I applied the optimized method to characterize irradiation-induced DNA damage for Familial Partial Lipodystrophy (FPLD), a rare genetic disorder that exhibits an enigmatic behavior under irradiation. My research focused on comparing the effects of Conventional-RT and FLASH-RT on cellular responses, using mutated cells from FPLD-afflicted patients and healthy samples. I conducted a quantitative analysis of repair foci, linking the signal obtained to the quantity of proteins recruited to damaged sites. Clustering algorithms were then applied to determine protein quantities within clusters, aiding in the analysis of histone averages and densities within repair spots.
Additionally, the study examined variations in response under different radiation doses and the unique behavior of cells with the FPLD mutation. This research advances the in vitro characterization of the FLASH sparing effect using advanced biophysical and imaging tools.
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